DP25 en

Manual
DP25
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ColorView III
DP25
The DP25 camera 3
SYSTEM REQUIREMENTS ......................................................4
Acquiring images 5
WHEN CAN COLORS APPEAR TO BE FALSE? .....................6

WHEN CAN THE IMAGE BE TOO DARK? ..............................6
CALIBRATING INTELLIGENT EXPOSURE .............................9
EXPOSURE TIME LOCK ........................................................10
Acquiring images with manual control 10
FURTHER OPTIONS IN THE CAMERA CONTROL ..............16
NetCam 23
INSTALLING THE WEB SERVER ..........................................24

NETCAM IN THE INTRANET .................................................24
NETCAM IN THE INTERNET .................................................24
USING NETCAM .....................................................................26
Setting up the camera 28

EDP250907
LOGICAL INPUT CHANNELS ................................................28

INPUT TAB ..............................................................................29
DISPLAY TAB .........................................................................31
FORMAT TAB .........................................................................35
ColorView III
Appendix I: Microscope settings 37
ILLUMINATION ....................................................................... 37
OBJECTIVE, TOTAL MAGNIFICATION AND USEFUL MAGNI-
FICATION ............................................................................... 39
Appendix II: How to install the camera 40
How to install the camera 42
When encountering problems with the cam-

era... 53
WEEE DECLARATION 55
CE Declaration of Conformity 56
The DP25 camera
DP25
The DP25 camera
DP25 is a quick, high resolution CCD color camera used for biomedical and
What is the DP25? material sciences applications. It was specially developed for acquisitions
using light microscopy, has an especially compact design, and can be
connected to all types of light-microscopes via the C-mount. Its high
dynamic scope enables it to acquire images which have very light and very
dark areas
Technical data DP25
Dynamic range 3 x 12 bit,
Exposure times 200 µs to 16 s
Frame rates 32 / s with binning 4
8.4 / s at full resolution
Resolution options 2560 x 1920,
1280 x 960,
3
854 x 640 or
640 x 480
Camera Mount Standard C-Mount
PC-interface Firewire (IEEE 1394)
DP25 • System Requirements
System Requirements
To properly use DP25, the following system requirements for your computer
and your image analysis system should be taken into consideration:
Hard- / Software Required
CPU P4 with at least 1,8 GHz
Memory 512 MB
Hard Disk Storage 30 GB
Graphics VGA with 32 MB integrated graphic
memory
Interface Firewire (IEEE 1394)
Operating system Windows XP Pro, SP2
Image analysis program analySIS FIVE, build 1175 or higher
cell^A, cell^B, celld^D, cell^F, cell^P,
build 1175 or higher
Overview of the chapters

"Acquiring images" on page 5 describes the automated acquisition of
images with the help of the Intelligent Exposure function. In the majority of
situations you will be able to use this method to acquire high quality images.
Should you not attain the required acquisition quality with Intelligent Expo-
sure, or should you basically prefer to work with manual controls, chapter
"Acquiring images with manual control" on page 10 describes how to work
with this method.
"NetCam" on page 23 describes the use of the NetCam add-in for making
direct transmissions of images via the Internet or an intranet.
In chapter "Setting up the camera" on page 28 you can learn how to change
the camera's settings, and how you can define different input channels for
different tasks.
"Appendix I: Microscope settings" on page 37 provides you with a short
overview of how you select the correct settings for your microscope in order
to achieve the best results.
4
"Appendix II: How to install the camera" on page 40 contains detailed

instructions about the installation of the camera.
Acquiring images
Acquiring images
Your image analysis program comes with a procedure for automatically
acquiring images. By using Intelligent Exposure you can acquire high quality
images with just a few mouse clicks.
1) Click the Intelligent Exposure button in the Standard button bar.
" Should you use Intelligent Exposure for the first time with this
camera, you will be asked to perform a calibration. (see Calibrating
Intelligent Exposure on page 9)
" The Intelligent Exposure dialog box will be opened.
2) Click the Acquire > button.

" The complete acquired image will be shown in the document
window. Check if the required image segment is shown in this
setting.
" The exposure times for the live-image and for a snapshot are
displayed in the dialog box's status bar.
3) Find the place on the sample that interests you.
4) Clicking the Focus mode button will enable you to bring the image into
5
focus.
" The central image segment will then be magnified to the point
where one camera pixel is equivalent to one pixel on the image.
This setting enables you to focus optimally.
5) Bring the image into focus.
" To help you do this, you can use the sharpness monitor.
6) Click the Focus mode button to return to the complete image.
7) Then click the Snapshot button to acquire a concluding image with the
settings you have chosen.
" This image will remain available to you even when the Intelligent
Exposure dialog box has been closed, while the live-image will be
deleted.
Related topics
Sharpness Monitor 20
DP25 • When can colors appear to be false?
When can colors appear to be false?

For the color reproduction, the correct settings for the camera's three color
channels, red, green and blue, are decisive.
When is it necessary to perform a white balance?

The most important criterion for the color settings of the camera is the
comparison between the image in the eyepiece and its display on your
monitor. A white balance can be made necessary by the following changes:
• The lamp is exchanged
• The lamp voltage is changed.
This is how you perform a white balance with Intelligent Expo-
sure
1) Procure a sample that is suitable for a white balance.
" For reflected light microscopy this will be a standardized gray card.
Should one not be available, you can also use white laboratory
porcelain or neutral ND-filter paper.
" When using the transmitted light method, look for a position at
which the light passes through the cover slip, embedding material,
and microscope slide, but not through the specimen. Should you
not find a position, remove the sample and perform the white
balance without a sample.
2) Position the sample under your microscope and start Intelligent Expo-
sure.
3) Click the White balance button to set an ROI (Region of Interest) for the
white balance with Intelligent Exposure, and to activate the white
balance.
" Pull the ROI's frame shown in the image document to the required
position and set its size by moving the mouse while keeping the
right mouse button depressed.
" Rightclick to confirm position and size of the ROI.
4) Then continue as usual.
6
The white balance will then remain activated until you once more click the
White balance button.
When can the image be too dark?

Very bright image segments or ones that reflect strongly, lead to a false
exposure time.
Intelligent Exposure calculates the exposure time that is necessary by
analyzing the signals. Normally, when this is done, all of the pixels will be
used. Very bright areas in an image, or areas that reflect strongly, cause the
exposure time to be reduced, to also enable these areas to be correctly
displayed. Areas that are not so bright, or don't reflect so strongly, will then
appear to be darker than they are.
When can the image be too dark?
This is how you avoid your images being made dark by bright
areas
In this example a graphite sample is to be acquired.
1) Start Intelligent Exposure and click the Snapshot button.
" The darker area of the sample will then be displayed very darkly.
The lower part of the image

shows a fiber.
In the default setting,
scratches and inclusions
don't stand out very much
from the background.
This makes an automatic
evaluation difficult.
2) Click the Set ROI for White Balance button.

" The mouse pointer appears in the image. On its tip, a red rectangle,
the so-called ROI (Region of Interest), can be seen.
3) Move the mouse to alter the position of the ROI. Change the size of the
ROI by keeping the mouse key depressed and moving the mouse.
4) Position the ROI in an image segment that you can display well.
" While you are moving the ROI, the exposure time will not yet be
changed.
5) Click your right mouse button to fix the size and position of the ROI.
" Intelligent Exposure will then readjust the exposure time to have the
area within the ROI optimally displayed.
7
When the ROI for the calcu-
lation of the exposure time is
moved to the darker area of
the sample, the sample and
its structures will be brighter,
and more clearly shown.
With these settings, an auto-
matic evaluation is made
much easier.
DP25 • When can the image be too dark?
This is how you avoid your images being made too dark by
strongly reflecting areas
In this example, a coin with strongly reflecting spots is to be acquired.
1) Start Intelligent Exposure and click the Snapshot button.
" The sample will be displayed very darkly, since the reflecting spots
simulate a bright sample area when the exposure time is calcu-
lated.
With the default setting of

the exposure time correc-
tion, the reflecting areas on
the surface of this copper
coin cause the complete
image to be shown darkly.
The histogram shows that
there are no bright pixels in
the image.
The camera's dynamic
scope will then not be fully
utilized.
2) Move the Exposure adjustment slide control to the right.

" The exposure time for snapshots will then be increased. The dark
areas will then become brighter.
Under the same conditions,
the Exposure adjustment
slide control has here been
moved to the right. This has
8
lead to a noticeable
improvement in the contrast.
Intelligent Exposure calcu-
lated an exposure time that
was twice as long.
The histogram shows that
the camera's dynamic
scope has been utilized
much better.
Calibrating Intelligent Exposure
Calibrating Intelligent Exposure

Before you use it for the first time, you have to calibrate Intelligent Exposure.
The calibration will be automatically set off when you start Intelligent Expo-
sure for the first time.
Intelligent Exposure arranges linear correlation between the exposure time
and the mean intensity for calculating the optimal exposure time. In order to
take advantage of this correlation, the camera's dark current must be known
as an offset of the linear function. Thus, your first step when working with
Intelligent Exposure will be to measure this dark current via the acquisition
of an image using a darkened camera. When first starting Intelligent Expo-
sure, you will be automatically asked to carry out this calibration.
This calibration step measures the camera's dark current, in order to make
an offset correction possible. After successful measurement of the dark
current, Intelligent Exposure asks you to take the next step.
The second calibration step consists of measuring the signals at various
9
camera resolutions. In this way the camera's sensitivity will be determined.
The image display changes considerably during this step.
Repeating the The calibration is specific to each camera.
calibration Should you change the camera, you will have to perform the calibration once
more. The Calibrate... button, located in the Intelligent Exposure > Options
dialog box, gives you access to this function.
DP25 • Exposure Time Lock
Exposure Time Lock

Exposure Time Click the Exposure Lock button in the Intelligent Exposure dialog box to lock
Lock the current exposure time. From this moment on, no more changes will be
made to the exposure time, even if you shift the Exposure time slide control.
Click the button anew to return to the dynamic adjustment of the exposure
time.
Use the Exposure Lock option when you want to compare the intensity of
different images or when you want to combine them. You will want to
combine separate images, for example, when you use the MIA add-in, with
which the images are combined into one composite image.
Acquiring images with manual control

The methods that have been described for making acquisitions using Intel-
ligent Exposure already cover a wide range of possible applications. In
special cases, or when you prefer to have full control over your camera, you
can apply manual controls.
The manual control of your camera offers you numerous possibilities. The
instructions that follow demonstrate a typical working plan. By means of
using different settings for the acquisition parameters you will be shown how
the possible options affect the results you achieve.
Preparing the input channel

1) Use the Image > Configure Input... command.
" The Configure Input dialog box will be opened.
2) On the Display tab select the Online histogram check box.
" The online histogram is an important tool that helps you adjust the
camera while the acquisition is being made.
3) In the Image scaling picklist, select the way you want the camera image
to be displayed on your monitor.
" For examining the sample, select the Adjust to viewport option.
With this setting you will see the camera's complete image on your
10
monitor while you are making the acquisition. It is suitable for

moving to the area of the sample that interests you.
" To focus, select the Full size (100%) option. With this option each
monitor pixel is equivalent to one camera pixel.
4) Click the OK button to close the Configure Input dialog box.
Acquiring a live-image
5) Click the Acquire button in the Standard button bar to start the acquisi-
tion.
" The live-image will be displayed in the image document.
" The online histogram will be displayed.
6) Click the Camera Control button in the Standard button bar.
" The Camera Control dialog box offers you all of the possibilities for
making settings for the camera.
11
At this point, the exposure time has not yet been correctly set. The histogram shows that the camera's dynamic range
has not been fully utilized. The image appears to be very dark.
DP25 • Acquiring images with manual control
Adjusting the exposure time

7) Click the Auto button, located in the Camera Control dialog box, to have
the exposure time automatically set correctly.
" Alternatively, you can change the exposure time by entering the
values you want, or by clicking the [+] and [-] keys. While doing this,
keep an eye on what the histogram shows.
8) Click the Snapshot button to acquire a snapshot with the current
settings.
12
The automatic calculation of the exposure time results in the image becoming considerably brighter. The camera's
dynamic range will then be more fully utilized.
Automatic Gain Display and fixed scaling

Even when the exposure time has been correctly set, it is still, in certain
cases, possible to improve the image quality. Which of the two methods,
automatic gain display and fixed scaling will achieve the best results,
depends on the sample. Try out both of the methods as described in the
following. The properties of both methods will be explained in the section
"Difference between Automatic Gain Display and fixed scaling" on page 15.
Automatic Gain Display

1) Click in the Camera Control or Histogram dialog box, the Use automat-
ic gain display button, to activate the Automatic Gain Display. The Au-
tomatic Gain Display regulates the amplification of the signal
continually, so that a fixed proportion will land in the overflow (Channel
0 resp. Channel 4095).
" You can set the proportion of the signal that is to land in the over-
flow by using the Image > Configure Input > Display command.
When you activate the Automatic Gain Display, the range that is displayed will be limited correspondingly to the signals
present. In the example shown, it reaches from channel 302 (blue line in the histogram) to channel 3505 (red line).
13
DP25 • Acquiring images with manual control
2) Click the Histogram calculation on ROI to limit the image segment on

which the display is to be optimized.
3) In the Camera Control dialog box, click the Set ROI for Histogram
button.
" A red rectangle will appear in the image. The rectangle indicates
the area that is to be taken into account for the histogram calcula-
tion. To change its size, move the mouse while keeping its left
button depressed, and to change its position move the mouse
without pressing its button.
4) Position the ROI in such a way as to encompass the area that is to be
displayed as well as possible.
14
Limiting the calculation of the histogram data to that of the red rectangle makes the image appear brighter. The calcu-
lation of the Automatic Gain Display is only based on the data from this rectangle.
Fixed scaling
1) In either the Camera Control or the Histogram dialog box, click the Use
fixed scaling button.
" The Automatic Gain Display will be deactivated.
2) Click the Automatically adjust fixed scaling button to have the once
more specified area adapted to the current conditions.
By having the fixed scaling automatically set, the range displayed will be limited according to the signals present. Only
15
the upper limit (red line) will be changed, in order to avoid color deviations.
Difference between Automatic Gain Display and fixed scaling

Automatic gain display and fixed scaling are both methods of transforming
the crude camera data into an image that can be displayed. While the
camera works with a resolution of 12 bit per color channel, for the display,
storage and further processing, only 8 bit per channel can be used.
Automatic Gain The Automatic Gain Display carries out the adaptation dynamically
Display according to an analysis of the histogram. In this way the image quality will
be automatically adapted when changes are made in the illumination or the
sample. It is, however, not possible to compare separate images that have
been acquired with this setting, where their intensity is concerned.
What counts for the Automatic Gain Display, is the area on which the histo-
gram has been calculated.
DP25 • Further options in the Camera Control
Button Area Effect

Full image The histogram is calculated on the complete image.
This option is recommended if the structures to be observed
are equally dispersed throughout the entire image.
Crosshairs The histogram is calculated according to two strips that run in
vertical and horizontal directions in the form of a cross, and
are only a few pixels wide.
ROI The histogram is calculated on a rectangular area that is de-
fined by clicking on the Set ROI for Histogram button.
This option is suitable when the reproduction of a part of the
image is to be optimized.
Fixed scaling With the fixed scaling option the setting that has been made will not be
altered. This makes it possible to compare the intensity of images that have
been acquired with the Fixed scaling option.
This makes it possible to define a fixed intensity range. This method is
mostly used for monochrome acquisitions.
Further options in the Camera Control

White Balance
The human eye reacts very sensitively to color variations which are
perceived as a color tingeing effect. The adjustment of the color channels
and the white balance depend on the illumination conditions. When you
change the lamp or altering the lamp voltage, you will have to subsequently
carry out another white balance.
When using white balance, the individual color channels are scaled in such
a way that the white or neutral gray area of the image displayed on the
monitor is displayed correctly as white or gray. The white balance can be
carried out automatically on each image or manually for individual images.
Microscope Settings The microscope settings you use for the acquisition for a white balance must
correspond to those you use in practice. This especially holds true for the
16
filter and the lamp voltage settings. The lamp voltage of a 12V halogen lamp
should be set to 9V. It should, however, not be set lower than 5V. Use the
color neutral ND-filter, should you have to reduce light intensity.
Specialized microscopy It is very difficult to execute a white balance in some microscopic methods,
techniques i.e. DIC or polarization. In these cases, execute the white balance using the
bright field method and only then begin with the special methods.
Transmitted Light When using the transmitted light method, look for a position at which the light
passes through the cover slip, embedding material, and microscope slide,
but not through the specimen. Should you not find such a position, remove
the sample and perform the white balance without a sample. When you do
that, the lamp light used for illuminating the specimen will fall directly on the
objective.
Reflected light When using the reflected light method, you need a reference surface. Stan-
dardized gray cards are best for this. Should one not be available, you can
also use white laboratory porcelain or neutral ND-filter paper. Should your
samples contain very reflective areas, defocus your microscope when
setting the white balance.
Performing a white balance with manual acquisition

1) Acquire an appropriate snapshot.
2) Use the Image > Camera Control... command.
" The Camera Control dialog box will be opened.
3) Click the Set ROI for White Balance button.
4) Pull the ROI's red frame, shown in the image document, to a position
which is a uniform white or gray.
" Change the size of the area by moving the mouse while keeping its
left button depressed. The size you can make this ROI for the white
balance is limited.
" The ROI can be difficult to see if the image has a reddish shade. In
this case, use the Special > Preferences > View command to
change the color of the frame. You can select a more appropriate
color from the Primary list located in the Mouse cursor group.
5) Rightclick to confirm position and size of the ROI.
6) Click the White Balance button to perform an automatic white balance.
" The settings for the individual color channels are optimized in the
set area. The result is directly visible in the image buffer's image.
7) Click the Color Settings button.
" The Color Settings dialog box will be opened.
8) Click the Define...button.
" The Define Default Color Settings dialog box will be opened.
9) Click the Current settings button to adopt the current values for the rest
of the session.
" When using this setting, the white balance is valid for all images
which are acquired with this input channel during the current
session.
10) Click the OK button to return to the Color Settings dialog box.
11) Click the Standard button to save these values as general standards.
" The current values for the white balance are now saved and will be
imported when the program is restarted. A manual change of the
color settings changes the white balance.
The image analysis program alters the color settings in such a way that the
17
image area in the ROI appears as white or gray as possible. After you have
carried out the white balance once, you can activate it at any time during a
running live-image acquisition.
Sharpen filter
Use the sharpen filter to increase the sharpness of your live image by using
edge enhancement.
Sharpen Filter Click the Sharpen Filter On/Off button to activate/deactivate the sharpen
On/Off filter. The manner in which the acquired image is affected by the sharpen
filter depends on the sharpen filter parameter. You can adjust this parameter
by using the Sharpen Filter Settings dialog box.
Clicking the Sharpen Filter On/Off button produces the same results as
selecting/clearing the Activate check box in the Sharpen filter group in the
Input tab. The button is simply a more convenient alternative.
Further information on sharpen filters and sharpen filter parameters can be
found in the section "Input tab" on page 29
Sharpen Filter Click on the Sharpen Filter Settings button to open a dialog box in which you
Settings can set the sharpen filter parameter:
To adjust the numerical value of the sharpen filter parameter, use your
mouse to move the slide control to the right or left. This is a convenient alter-
native to using the Input tab - where you can also adjust this numerical value
(in the Sharpen field).
Color Settings
Click the Color Settings button to manually alter the camera's color settings.
The settings can be altered by using the five slide controls: Red, Green,
Blue, Gamma, and Saturation.
Red, Green, Blue The Red, Green and Blue slide controls can each be set from 0,00 to 20,0.
18
The set value is the factor with which the signal of the corresponding color
channel is multiplied.
Gamma Use the Gamma slide control to obtain a nonlinear contrast enhancement.
Please note that this gamma filter does not only affect monitor display but
also alters actual image information. If you are planning on quantitatively
evaluating your images at a later time, use the value 1.0 for the gamma filter.
The Oper menu provides you with commands for adjusting image contrast
at any later time.
What is gamma? As is the case with the slide controls of the individual color channels, the
numerical value for gamma effects how the illumination intensity of the
pixels of the camera CCD chip is converted into the color value for the corre-
sponding pixel in the image buffer. If gamma equals 1, the conversion from
illuminance into gray/color value is linear. If gamma does not equal 1, this
results in a nonlinear contrast enhancement. If gamma is smaller than 1, the
brighter gray values will be spread more significantly than the darker ones.
The result will be enhanced contrast in the lighter areas of the image. The
image as a whole will become darker. If gamma is greater than 1, the darker
gray values will be spread more significantly than the brighter ones. The
result will be enhanced contrast in the dark image areas. The image as a
whole will become lighter.
Saturation The Saturation slide control enables you to alter the color rendering from a
black and white signal to maximum color saturation. In most cases, the
"good" setting of this value is seldom higher than the range of 0,10 to 0,20.
The Gamma slide control's value range is from 0,00 to 2,00; the Saturation
slide control's range is from 0,00 to 0,99.
Contrast You can change the image contrast by using the Contrast slide control.
Move the slide control up to increase the contrast of the image. Move the
slide control down to reduce the image’s contrast.
Possible values for the Contrast slide control range from 0.00 to 2.00. A
value of 1.13 is preset. The contrast value is a relative, nondimensional one.
Brightness With the Brightness slide control you can manipulate the image’s brightness.
Move the slide control up to increase the brightness of the image. Move the
slide control down to reduce the brightness of the image.
Brightness and Auto- If you change the Brightness slide control while the Automatic Gain Display
matic Gain Display is activated, the upper limit set for the right overflow of the Automatic Gain
Display will be disregarded.
Default Click the Default button to use the default values defined in the Define
Default Color Settings dialog box from now on.
Define… Click the Define... button to set the standard color settings. The Define
Default Color Settings dialog box will be opened.
19
Factory settings Click the Factory settings button to return the color settings to their original
levels. The factory settings have been adapted in such a way that the color
rendering with a specific equipment configuration is ideal for a wide range of
applications without any additional changes having to be made.
Current Settings Click the Current settings button to adopt the current values as the future
standard.
Related topics
Automatic gain display 32
Sharpness Monitor
The Sharpness Monitor allows you to control the focus settings during a live
acquisition. You are also able to define a rectangular area of the image
which is to be analyzed for the sharpness monitor in the same way as for the
white balance and black balance. The maximum contrast serves as a
monitor for the sharpness in this area. Therefore this area should be limited
to an image segment in which the contrast is a good monitor for the sharp-
ness.
The Sharpness Monitor consists of a dialog box in which a relative measure-
ment of the sharpness is indicated by a moving bar that varies between
Blurred and Focused.
If you start a live acquisition with an activated Sharpness Monitor or activate

it during a live acquisition, it will show middle sharpness (50%) in all cases.
This is because there is no absolute measurement for the sharpness in an
image and because the sharpness monitor only gives relative values which
are related to the initial situation that was present when the live acquisition
was started.
Focussing and Sharp- The length of the bar changes if you alter the focus settings during acquisi-
ness Monitor tion with an active sharpness monitor. In doing so, the blue marking bar
shows the maximum sharpness reached since the live acquisition was
started. The bar shifts accordingly to the right should the sharpness increase
due to your changes. If the focus settings were already optimal when the
acquisition was started, the bar can only shift in direction Blurred. The lowest
sharpness value will be indicated by a green marking bar.
Reset to 90% Click the Reset to 90%button to set the sharpness monitor's relative value
to 90%. Utilize this function if you start working with a very blurry image and
if the sharpness monitor's bar reaches the limit to the right while you focus.
By resetting to 90%, the display gets more room for additional focussing.
20
The Reset to 90% function is also available if the sharpness drops very dras-
tically due to changes made in the microscope settings (i.e. changing objec-
tives) during an acquisition in progress.
Using the sharpness monitor

1) Set all other parameters (microscope settings, exposure time, white
balance) optimally. The application of the sharpness monitor should be
the last step before image acquisition.
2) Acquire a snapshot.
3) Use the Camera Control command.
4) Click the Set ROI for sharpness monitor button to interactively define a
rectangular area of the image for the sharpness monitor.
" To do this, select an area of the specimen for which a good sharp-
ness setting is especially important.
5) Begin a live acquisition.
" The Set ROI for sharpness monitor button becomes inactive.
6) Click the Sharpness Monitor On/Off button to open the Sharpness

Monitor dialog box.
" The sharpness monitor begins with a value of 50%.
7) Vary the focus settings and observe the sharpness monitor while doing
so.
" The sharpness monitor bar shows the current sharpness, the blue
mark shows the highest achieved sharpness.
" Pay attention to the exposure time while you set the focus.
Changes in the sharpness monitor occur with a certain amount of
delay when you use longer exposure times (> 100ms). Vary the
focus settings in small intervals and wait each time until the sharp-
ness monitor adapts itself as well.
8) Find the focus settings by which the sharpness monitor is at its
maximum and use these settings for the acquisition.
Partial Readout
The "Partial Readout" mode is a method of making the live-mode react more
quickly. For the "Partial Readout" mode, a rectangular part of the image is
defined, to which the readout is limited during the live acquisition. In this way
a higher frame rate is attained, since a smaller amount of data has to be
transferred.
The live overlay must be enabled for the "Partial Readout" mode. This takes
place on the Display tab in the Configure Input dialog box.
Partial Readout and Clip- "Partial Readout" mode and clipping function both lead to smaller images
ping than those possible with the camera. The "Partial Readout" mode already
goes into action while the image is being read out from the camera. The
parts of the image that are removed by this function will not even be
exported to your computer. In contrast, a clipping will not be calculated until
all of the image data has reached your computer. This means that clipping
does not accelerate the image acquisition.
Owing to the camera's very high pixel count, when full resolution is being
used, the frame rate is often not high enough to enable good focussing
during live-mode.
21
Clipping and the "Partial Readout" mode are mutually exclusive. The "Partial
readout" mode is disabled if you enable a clipping on the Format tab located
in the Configure Input dialog box. The ROI which was set for the "Partial
Readout" mode remains intact. Clipping is disabled if you enable the "Partial
Readout" mode while clipping is enabled. The settings for the clipping will
be preserved. They will be reemployed as soon as you deactivate the
"Partial Readout" mode.
Related topics
Display tab 31
Using the Partial Readout

1) Start the live acquisition using full resolution.
2) Set the ROI for Partial Readout on an important image detail.
" Change the size of the area by keeping the mouse button
depressed while you move the pointer.
" Change the position of the area by moving the mouse with the
button released.
" Fix the area with a right click
3) Activate the Partial Readout.
4) Make the required settings.
5) Activate the Partial Readout mode.
" The image area set with the ROI for Partial Readout applies to the
following snapshot, should the Partial Readout mode be enabled.
6) Acquire a snapshot.
Black balance
The black balance is used for background correction. In an area (ROI) which
you have defined, your image analysis program calculates the mean value
for each color channel. These values are subtracted from all of the pixels in
the entire image. The black balance is to be used predominately for fluores-
cence acquisitions, where it is applied to remove undesired background
from the image. A discoloration of the images occurs if the black balance is
used with brightfield acquisitions.
Using the black balance

1) Acquire a snapshot containing an area that is suitable to be used for a
black balance.
2) Use the Image > Camera Control... command.
3) Click the Set ROI for Black Balance button to set an area in the image
in which the black balance is to be defined.
4) Drag the red frame that defines the ROI in the image document, to a
22
position containing an area which is as uniformly dark as possible.

" Change the size of the area by moving the mouse while keeping its
left button depressed. The size you can make this ROI for the black
balance is limited.
5) Rightclick to confirm position and size of the ROI.
" The values for the black balance will then be calculated on the area
you have thus defined.
6) Click the Black Balance On/Off button to activate the black balance.
" The correction values calculated for the black balance will then be
deducted from all of the pixels in the entire image.
The black balance function is available for live acquisitions as well as for
snapshots. A ROI for black balance has to be set as described above before
you perform the black balance on snapshots.
The Black Balance button for snapshots is located in the Black button bar.
Should the Black button bar not be on display, use the Special > Add-In
Manager command to activate it.
NetCam
NetCam
You can export the DP25 live-image to other computers via an intranet or
the Internet, by using the NetCam add-in.
This enables you to make the images immediately available to your
colleagues during an acquisition. This makes it possible for you to work
interactively over large distances by using simultaneous communication,
e.g., telephone. The images will be compressed in the process to speed up
the rate of transmission.
Transferred images The live-image will be directly transferred. Provided your network connec-
tion achieves an adequate speed and the frame rate attained during acqui-
sition, is sufficient, you will get the impression of a live-image. In an internal
company network, an immediate feedback without any time delay will be
possible between observer and user.
NetCam can also transfer a fixed image, whereby in this case the image's
overlay will also be transferred.
What is a Server? A Server is the computer on which both the image-analysis program and the
NetCam are installed. The image acquisition is controlled from the server.
What is a Client? A Client is any computer that is able to access the server's images via the
Internet. The client can’t control image acquisition.
To make images available via the Internet, a web server must have been
installed on your computer. An appropriate program will be installed during
the setup, if you have selected the NetCam add-in. This step will be skipped
should a web server already exist on your computer.
Warning When you use a computer connected to a network it is possible that the
security settings will not allow its use as a web server. In this case, contact
your system administrator.
In your image analysis program's Module\NetCam subdirectory you will find
the file "NetCam_Info.pdf". This file contains technical explanations about
NetCam which you can give to your system administrator.
23
DP25 • Installing the web server
Installing the web server

The web server you require for using NetCam will be automatically installed during the setup.
The installation of the web server is started directly as part of the setup. After
you have agreed to the license terms, you can make further settings in a
dialog box.
You can simply adopt the default settings.

Complete, Custom We recommend the complete installation of the web server.
NetCam in the intranet

24
The transfer of images within the company network or intranet generally

does not require any further actions.
Enabling access During installation, the setup will create a link with the name NetCam.html in
the \Module\NetCam subdirectory below your image-analysis program's
working directory. Send this link via E-mail to colleagues you would like to
give access to your images. The easiest way to do this is with the following
command: Send to > E-mail receiver in the context menu of the link in
Windows Explorer.
Prerequisites for clients Java Runtime Environment, Version 1.4.1_03 or higher is required to show
the images on the clients. When this requirement has not been fulfilled by
the client, he will not be able to receive the images.
NetCam in the internet

When using NetCam, a direct communication takes place between the
server and the client. If the communication goes beyond the company
network, you must, if need be, make changes to the security settings.
NetCam in the internet
Adapting the security The adaptation of a network's security settings takes place via the respec-
settings tive system administrator. The necessary technical and security require-
ments for the use of NetCam are outlined in a separate file entitled
"NetCam_Info.pdf" which will have been saved during setup to your image
analysis program's \Module\NetCam subdirectory. Please give this file to
your system administrator so that he can make the appropriate adaptations.
IP Address The server's internet address must be entered on the client's page. This
address is available in the well-known alphanumerical form ("MyCom-
puter.OurCompany.de") and in the numerical form ("62.180.61.137"). The
numerical form of the address is more reliable, especially if your system
administrator has to alter the network settings.
Determining the IP You can find out your computer's IP address from your system administrator
address or determine it yourself.
Determining the IP address

1) Select the operating system command Start > (All) Programs > Acces-
sories > Command Prompt.
" The Command Prompt console box will be opened.
2) Enter the ipconfig command via your keyboard and use the Enter key.
" Data about your computer's network connection are shown in the
console box.
The ipconfig command will
display a computer's IP
address.
25
3) Note the entry in the IP address row.
4) Supply the users who are to have access to your server as NetCam
clients with this IP address.
Prerequisites for clients Java Runtime Environment, Version 1.4.1_03 or higher is required to show
the images on the clients. When this requirement has not been fulfilled by
the client, he will not be able to receive the images.
DP25 • Using NetCam
Using NetCam
The use of your image-analysis software remains the same when you use
NetCam. You can use all of the functions as always. To transfer the images
to other users and to be able to control the transfer, use the buttons located
in the NetCam button bar.
NetCam on/off Click the NetCam On/Off button to start or end the transfer of images with
NetCam.
The transfer of live-images begins as soon as NetCam is activated. The
transfer continues until you deactivate NetCam or close your image-analysis
software.
NetCam Click the NetCam Settings button to set image options and to administer the
Settings clients which are to have access to your images. The NetCam Settings
dialog box will be opened.
Connections
Use the Connections tab to administer the clients which are to access your
images.
26
Connected Clients In the Connected Clients list you will see all of the clients who are currently
connected to your computer. You can mark one of these clients to discon-
nect the connection to it or to refuse a connection in the future.
Disconnect Click the Disconnect button to disconnect the connection to the marked
client. The client can reconnect later.
>> Click the >> button to include the marked client in the Banned Clients lists.
The corresponding client will be automatically disconnected.
Banned Clients In the Banned Clients list you will see all of the clients who you have
forbidden access to your image-analysis software. From these computers it
is impossible to load and view the images made available by NetCam.
Add new... Click the Add new... button to add computers or entire groups of computers
to the Banned Clients list. The Enter IP(s) or a domain dialog box will be
opened.
Using NetCam
Domain name Enter the alphanumerical name of a domain in the Domain name field if you
want to block the access of all computers belonging to a certain domain. For
example, enter the domain name foobar.com to block the transfer of the
images to all of the computers within the foobar.com domain.
Procedure NetCam sends a query to the name server to get the numerical IP addresses
belonging to the specified domain. Should the query lead to an error, you will
receive a message. The specified domain will, in this case, not be placed on
the Banned Clients list.
IP Address(es) You can provide a numerical IP address or an entire address range in the IP
address(s) group to exclude the computers in question.
From, to Enter the limits of the address range to be excluded in the from and to fields.
27
Image options
In the Image Options tab you can set the quality of the transferred images
and thus influence the transfer rate.
Image quality Use the Image Quality slide control to control the compression of the
images. Move the slide control to the right to maintain a high image quality.
The data amount of the transferred images increases, whereby the transfer
rate decreases. Move the slide control to the left to increase the compres-
sion. The image quality decreases, whereby the transfer rate increases.
Image quality and data A single image of DP25 can have the size of up to 15 Mbyte. With an image
transfer rate of this size, the transfer to a client can take a long time, even if you're using
a very quick network connection; the effect of a live-image being transferred
will be lost. For this reason, images are compressed for transfer. The algo-
DP25 • Setting up the camera
rithm is equivalent to that used with JPEG images. Thus, high compression
rates are achieved, whereby, however, image quality decreases.
The amount of compression achieved is based on the image type. Images
with large, uniformly colored areas can be compressed more than detailed
images without these characteristics.
Setting up the camera

Logical input channels
For every camera that is to be used with the image analysis program, one
or more logical input channels will be set up. Logical input channels contain,
for example, the information about how large an image format is, and what
is to happen with the image signal before it reaches the image buffer. Each
channel must be calibrated separately. A channel for image acquisition via
DP25 is usually called "DP25 FW...".
Status after installation An input channel is automatically created when the software is installed,
immediately enabling you to make image acquisitions.
Precalibration During the installation you will be offered the opportunity of performing an
XY-calibration by entering the objectives and the TV adapter to be used.
Thanks to this automatic calibration you can then acquire calibrated images
without any further preparations.
Configuration via tabs Input channels are configured via the inputting of suitable values in various
tabs. The 3 tabs relevant for DP25 will be dealt with in-depth.
Setting up additional You can set up and use more than 100 additional input channels. This can
input channels be useful when you want to employ different settings for a variety of uses. If
you have to set up a new input channel, you can create a copy of an already
existing input channel and adapt it, or create an entirely new input channel.
Set Input Use the Image > Set Input... command to select a new input channel or to
create a new one. You can also depress the [F6] key. The Set Input dialog
box will be opened.
The Set Input dialog box
with an already defined
28
input channel.
New Channel Click the New Channel button in the Set Input dialog box to create an entirely
new input channel. The Select device dialog box will be opened. All
connected image creating devices are listed in the Available devices list.
Input tab
Select Device dialog box

with a connected DP25
camera.
Mark the DP25 in the Available devices list and click the OK button, in order
to define a new input channel.
Creating additional input Should you want to create additional input channels, for example to have
channels different configurations immediately available for repeating tasks, use an
already existing input channel.
Duplicate Click the Duplicate Channel button to make a copy of the active input
Channels channel. The new input channel gets the same name as the channel from
which it was copied and receives a sequential number. The newly created
channel is automatically enabled.
Configure Click the Configure Input button to adapt the settings for the new channel.
Input The Configure Input dialog box will be opened. You can also open this dialog
box by using the Image > Configure Input command or by double clicking
the camera name located in the status bar.
Input tab
29
DP25 • Input tab
Exposure Enter the number of microseconds the camera's CCD chip is to be exposed
in the Exposure field. You can also use the arrow key (to the right of the field)
to select this value. How you can change the value in this field with the help
of the Camera Control... command interactively while viewing a live-image
is shown in the section "Further options in the Camera Control" on page 16.
The shortest possible exposure time is 100 µs and the longest is 160 s.
Resolution You can choose from four different resolution possibilities for DP25 from the
Resolution list.
Sharpen filter Select the Activate check box in the Sharpen filter group to enhance the
image sharpness. This takes place in real time during image acquisition.
When applying the sharpen filter, the numerical value in the Sharpen field
determines whether or not the image’s contours appear sharper or softer.
Raising sharpness will accentuate edges, but will also increases image
noise. Reducing the sharpness makes the image softer. If you enter a value
of 0, the sharpen filter will have no effect at all.
Please note that when you use the sharpen filter you not only alter the
monitor display, but the actual image information as well. This means that
when you are planning on quantitatively evaluating your images later, you
should not use the sharpen filter. The Oper menu provides you with
commands for adjusting the sharpness of an image at any later time.
Convenient alternatives for activating/deactivating the sharpen filter, as well
as for adjusting sharpen filter parameters, are the Sharpen Filter On/Off
button and the Sharpen Filter Settings button - both in the Camera Control
dialog box.
Mirror The Mirror group enables you to determine if the camera's image is to be
mirrored directly into the image analysis program during transfer. Both
mirror functions can be combined with each other.
Horizontal Select the Horizontal check box to mirror the image horizontally during
acquisition.
Vertical Select the Vertical check box to mirror the image vertically during acquisi-
tion.
Info Click the Info... button to have a variety of device data displayed in the DP25
FW Information window.
30
Related topics
Acquiring images with manual control 10
Display tab
Device Name The Device Name relates to the inner construction of the camera and is
specified by the manufacturer.
Devices ID The Device ID is the physical identification of the camera for the operating
system. It is in accordance with the device's serial number.
Firmware Version The Firmware is the cameras software for your operation.
Serial Number The serial number serves as a clear identification for each camera
produced.
Camera Type The Camera Type is the exact identification for the camera model.
DLL Version The DLL Version is the version number of the camera's driver.
SYS Version The SYS Version is the version number of the camera's device driver. DLL
and SYS Version are identical.
Display tab
31
Over exposure The Over exposure group enables you to determine if and when you should
be warned of an overexposure. The software continually checks the image
intensity in live mode, by analyzing the image's gray-value histogram in real
time. You will be warned if the live-image is overexposed. The warning will
also appear following acquisition of an overexposed single image.
Display warning Select the Display warning check box to have a warning message shown if
the image is being overexposed:
In this case, reduce the exposure time until the warning message disap-
pears.
Overflow An image is overexposed when a certain percentage of its pixels attain the
maximum gray value possible. This percentage is entered into the Overflow
field.
DP25 • Display tab
Automatic gain display Use the functions in the Automatic gain display group to have the image
always displayed optimally on-screen, no matter what the actual lighting
conditions are. The system assesses the current gray-value histogram - in
real time - in order to obtain optimal display of the live-image on the monitor.
Please note that the Automatic gain display not only alters the monitor
display, but the actual image information as well. If it is activated, the reduc-
tion of the acquired image from 3 x 12 bit to 3 x 8 bit takes place with the
help of optimized lookup tables. If the Automatic gain display is not acti-
vated, the 4 lower bits (from the 12 bits of each component color) will be
clipped.
Activate Select the Activate check box to enable the Automatic gain display. The
effect of this check box is the same as clicking the Use automatic gain
display button located in the Camera Control dialog box (<). The Activate
check box is not available if the check box with the same name in the Fixed
Scaling group is enabled.
Right overflow When the automatic gain display is active, image intensity for the image on
the monitor will be spread between the gray values Gmin and Gmax. You
determine the values of these Gmin and Gmax gray values in the Left over-
flow and Right overflow fields: this is where you enter the percentage of
pixels with the lowest gray values which are to be displayed black, and which
percentage of the highest gray values are to be displayed white.
This is how you clip gray values that are to be ignored at both the upper and
lower ends of the gray-value histogram. Make use of the overflow values to
stop individual pixels or minute image areas that are too bright or too dark
from influencing the Automatic gain display.
Fixed scaling Instead of using the Automatic gain display group, you can use the Fixed
scaling group. The group whose Activate check box has been selected will
be the one available - the other will not be available.
The functions of the Fixed scaling group are for manually influencing on-
screen image display. You select definite limits - Gmin and Gmax - for the
gray-value range that you wish to have optimally spread for on-screen
display. Any gray values occurring outside this defined gray-value range will
appear either black or white.
32
Application This is how you can, for example, improve the visibility of the specimen
structures of interest to you - by having dark areas of the specimen
displayed entirely in black.
Please note that this manual presetting of the gray value, like the Automatic
gain display, not only affects the monitor display, but also the image infor-
mation.
Activate Select the Activate check box to switch on the manual definition of the gray-
value range to be displayed.
Right limit Enter the value of the right limit for the fixed scaling in the Right limit box. Its
function is equivalent to the Right limit parameter, which you can influence
by clicking the Manually adjust fixed scaling button located in the Camera
Control dialog box.
Display tab
Online histogram Select the Online histogram check box to enable you to keep an eye on the
gray-value distribution during image acquisition. During image acquisition, a
window showing the current histogram will appear automatically. This histo-
gram will be continually updated.
Online histogram during a
live acquisition The Fixed
Scaling mode is active.
Minimum, mean, and maximum gray values of the image will be shown
beneath the histogram. Percentages are in relation to the maximum obtain-
able gray value. The maximum obtainable gray value when using DP25
(with its view 12-bit depth) is 4095. This intensity corresponds to 100%.
During a live acquisition, you can turn the Automatic Gain Display on or off
with the buttons in the online histogram, as well as activate and set the fixed
scaling.
Setting the fixed scaling You use the mouse to set the limits of the fixed scaling in the online histo-
via the online histogram gram. Move the pointer over the blue (lower limit)or red (upper limit) vertical
line in the histogram. The pointer turns into a horizontal double arrowhead.
Move the mouse with the mouse button depressed to the left or right to either
increase or decrease the limit values. The changes can be viewed directly
in the live-image.
Live overlay Select the Live overlay check box if you want to use overlays in live-images.
Live overlays are absolutely necessary for the "Partial Readout" mode and
white balance.
An image’s overlay contains information in the form of text, markers, or other
graphical elements. The overlay can be displayed together with the corre-
sponding image without the actual image information being affected.
33
The Live overlay check box is only available if your display board features a
so-called hardware overlay which is supported by Microsoft DirectX. Almost
any up-to-date AGP display board provides you with that feature. You can
look at the System Info dialog box to check whether your display board
offers this feature. To do so, select the Info... command from the ? menu and
click the System Info... button. the System Info dialog box will be opened.
Click on the "DirectX" entry in the list on the left-hand side to have the
required information appear in the list on the right-hand side.
Displaying the histo- Select the Show histogram ROI check box to have the ROI for the histogram
gram ROI displayed during the live mode.
Image scaling Select one of the four selections for displaying an acquired image in a View-
port in the Image scaling list.
Related topics
Automatic Gain Display and fixed scaling 13
Partial Readout 21
DP25 • Display tab
Underscan Select the Underscan entry to have the whole image displayed. The system
will select the highest zoom level - 25%, 50% or 100% - where the whole
image can still be displayed within the Viewport. In certain cases not all of
the Viewport’s available area will be used.
Overscan Select the Overscan entry to ensure that all the Viewport’s available area will
be used. The system will select the lowest zoom level - 25%, 50% or 100%
- where the image fills the whole Viewport. In certain cases parts of the
image will not be visible.
Adjust to viewport Select the Adjust to viewport entry to have the image size adjusted to opti-
mally fit the Viewport. The image will be adjusted to fit the current size of the
Viewport.
Full size (100%) Select the Full size (100%) entry to have the image displayed without any
zooming. Only the left upper corner of the image will be shown if the View-
port is smaller than the image.
Histogram calculation Select the pixels to be taken into consideration for calculation of the gray-
value histogram in the Histogram calculation list. There are three options:
Full image If you select the Full image entry, the gray values of all pixels will be applied
to histogram calculation. This is the most commonly used setting for light
microscopy.
Crosshairs If you select the Crosshair entry, only pixels of a particular image area are
used in the calculation. This image area is shaped like cross hairs that are
centrally positioned on the image. The vertical and horizontal segments of
these cross hairs are respectively just a few pixels wide and cover just about
the entire height and width of this image.
ROI Select the ROI entry to carry out the optimization of contrast and exposure
time based on a rectangular frame.
These settings affect all of the real time functions that use the gray-value
histogram: the warning for Over exposure, the Automatic gain display and
the Online histogram.
Application We recommend selecting Crosshair if you wish to speed up histogram
calculation. You can speed up the frame rate of the live-image depending on
what real time functions you have activated. You should however make sure
that the pixel gray values around the middle image line and around the
34
middle image column are representative for the gray values of the entire
image. If this is the case, you will not notice any difference compared to the
Full image option for the over exposure warning, Automatic Gain Display,
and the online histogram.
Format tab
Format tab
Active area The display in the Active area field shows how large the acquired image
segment is compared to the largest possible image, when the current
settings are applied. If you make changes to the clipping, you can directly
view their effect here on the active area.
Image format you can select a resolution from the Image format list. The entries in this list
correspond to predetermined settings for the clipping. In addition to the
predetermined list elements, you can also select from the settings you have
defined.
Image resolution and With the DP25 you can influence the size of the images in pixels in two ways.
Clipping In the Image format list the sizes of images that can be achieved by cutting
off the edges (Clipping) are contained. In the Resolution list located on the
35
Input tab you can select resolutions that achieve the same image format in
pixels with a constant display area.
Binning The Binning function is not enabled when you use the DP25. Use the Reso-
lution function located on the Input tab instead.
Clipping border (pixel) When clipping is used, the borders of the images are cut off. Hence, clipping
does not have any effects on the resolution and brightness nor on the
required exposure time.
Left, Right, Top, The fields entitled Left, Right, Top and Bottom show the number of pixels
Bottom that are to be clipped from each image border. Each individual value can
range from 0 (no clipping) to almost the entire image in each respective
direction (complete clipping). If a value for the clipping has already been set,
the setting possibilities for clipping the opposite border reduce themselves
respectively.
Interactive Click the Interactive button to be able to place the clipping interactively in the
image document. Move the pointer with the mouse button depressed to
change the size of the area to be acquired. The currently acquired area will
Related topics
Resolution 30
DP25 • Format tab
be indicated by a red frame. Move the mouse to alter the position of the
marked area within the image document. Rightclick once to fix the area to
be acquired.
No Clipping Click the No Clipping button to return the values for Left, Right, Top and
Bottom to 0, thus disabling Clipping.
36
Appendix I: Microscope settings
DP25
Appendix I: Microscope settings

Your digital camera can only achieve high image quality if the microscope
has been optimally set. A subsequent correction to the software can never
correct deficiencies in the image resulting from faulty settings made to the
camera and microscope.
The optical system microscope offers numerous setting possibilities. In
order to attain best acquisition results, the settings must be made precisely.
In spite of varying models and observation possibilities, the microscope's
image producing components are functionally identical.
A microscope basically consists of the optical components: projection objec-
tive and magnifier. The projection objective is the objective with which the
observation is to be carried out. The eyepiece takes on the function of the
magnifying glass. An additionally important component is the illuminator.
This illustration clarifies the
basic assembly of the Camera
Kamera
Camera
components of a micro-
scope in transmitted-light Lupe
Lens
mode.
Okular
Ocular
Objektiv
Objective Projektion
Projection
Probe
Specimen
Kondensor
Condenser
Beleuchtung
Illumination
Lampe
Lamp
37
Illumination
Light is a decisive medium for creating images with a microscope. As a result, the light, i.e. illumination,
plays an important role in creating the image in the microscope.
The goal of the illumination settings is the equal distribution of light on the
sample with parallel light rays falling on the object. The illumination must be
bright enough to make all of the image details visible; however over-illumi-
nation leads to images being too bright and low-contrast. In addition, the
color of the light has a strong impact on the sample's color rendering.
Lamp Selection Tungsten-halogen lamps and gas discharge lamps are the most commonly
used lamps in microscopy. The light creating processes of both lamp types
are fundamentally different.
Tungsten-halogen Tungsten-halogen lamps comply in their functionality with a conventional
lamps light-bulb. Electricity flowing through a metal wire heats the wire to the extent
that light is produced (thermal emitter). In doing so, the wire emits a contin-
DP25 • Illumination
uous spectrum, hence a broad spectral band is emitted. One can control the
brightness as well as the color (color temperature) of the lamp by controlling
the electrical current. Tungsten-halogen lamps are widely used in micros-
copy, especially for acquiring color images.
Gas discharge lamp Gas discharge lamps stimulate gas at the atomic level using strong electrical
fields. The excited atoms return to their initial state and, in doing so, give off
the energy which initially charged them in the form of electromagnetic radi-
ation: light. The resulting photons each have characteristic energy which is
typical for the gas being used. As a result, this type of lamp does not emit a
broadband spectrum like the halogen lamp, but instead emits discrete wave-
lengths. This fundamental physical process cannot be externally influenced;
hence, such a lamp's perceived color is independent of the operation condi-
tions. Because of this discrete spectrum, gas discharge lamps do not emit a
color-neutral illumination. Its adjustable high intensity light and the emission
of short-wave spectral bands required for fluorescent microscopy is an
advantage.
ND filter Many microscopes can insert ND filters into the light path, thus reducing the
intensity of the lamp's light. These ND filters ensure a uniform reduction of
light intensity throughout the entire frequency range without altering the
color temperature.
Köhler Illumination
This illumination process which was described by Dr. August Köhler as early
as 1893, is still an indispensable prerequisite for successful analysis by light
microscopy. The Köhler illumination enables an optimal, homogenous illu-
mination of solely the part of the sample that is to be analyzed.
Condenser The condenser is responsible for the sample. The main components are the
aperture stop, with which the size of the illuminated area is set, and the
condenser lens, which is responsible for the parallel emitted light rays. The
condenser can be opened and closed. The condenser is height-adjustable
and horizontally adjustable.
Note Set the aperture stop to 2/3 of the entire opening and do not use it to reduce
the brightness. To do that, you should always use ND filters or a lower lamp
38
voltage.
Principle of Köhler Illumi- You will find comprehensive descriptions of the principle of Köhler illumina-
nation tion in microscopy text books. We will only give you a brief overview of some
important settings you should know.
Focussing the Field One vital step when setting up the Köhler illumination is achieving a sharp
Diaphragm view of the field diaphragm at the specimen level.
1) Focus the object to be viewed.
2) Close the field diaphragm so that only a small bright spot is visible in
the eyepiece.
3) Focus the image in the field diaphragm by using the condenser's height
adjustment.
Objective, Total Magnification and Useful Magnification
Adjusting the The illuminated has been optimally set.

Condenser 1) Open the field diaphragm, but only until its edge is still completely with-
in the visible area.
2) Center the image of the field diaphragm using the condenser's adjust-
ment screws.
" A precisely adjusted condenser guarantees that only that area of
the specimen you are examining is illuminated.
3) Repeat steps 1) and 2), until the edge of the field diaphragm is no
longer visible in the visual field.
Adjusting the Lamp You can generally skip adjusting the lamp with modern microscopes,
provided that the lamp is located in a frame and the bulb is in a centered
socket.
Objective, Total Magnification and Useful Magnification

There are no general rules for which objective you select; one should always
keep the specimen and reason for observation in mind.
Terminology The two most important parameters of an objective (magnification and
numerical aperture) are engraved in the objective. First the magnification
and then the numerical aperture which is separated by a diagonal slash.
Hence, the code 40 / 0,65 marks an objective with a magnification of 40X
and the numerical aperture of 0.65.
Magnification An objective's engraved magnification is of considerable importance for the
total magnification. However, one should note that the range of useful
magnification should not be surpassed.
Numerical Aperture The numerical aperture is the characteristic of an objective which deter-
mines its capacity. The numerical aperture can also be found engraved in
the objective. It is based on the refraction index of the lenses being used, as
well as the objective's angular aperture. Where the refraction index is
concerned, the manufacturers have to abide by narrow limits. To achieve
this, a fluid with a high refraction index is placed between the cover slip and
the objective. The immersion objectives made for this technique are marked
accordingly. They should be used with the appropriate immersion fluid only.
39
Total magnification The system's total magnification, to which an installed TV adaptor contrib-
utes, is based on the actual size of the final image. This can be the image
on the monitor, as well as the image printed on paper. To determine the total
magnification, it is best to use a stage micrometer which can be shown on
the monitor or the printout. You can determine the total magnification by
dividing the length of the object shown by the set length on the micrometer.
Useful magnification You can determine the total magnification by dividing the length of the object
shown by the set length on the micrometer. The useful magnification of the
optical systems microscope depends on the numerical aperture being used
( engraved on the objective). The useful magnification is in the range of 500x
to 1,000x the numerical aperture.
Empty magnification A loss of contrast and sharpness is the result if total magnification is larger
than the useful magnification. Check to see if replacing the TV adapter with
a 0,5 or 0,63 model sufficiently reduces the total magnification. If this is not
possible, you can also use an objective with the same magnification, but
with a higher numerical aperture.
DP25 • Appendix II: How to install the camera
Monitor Settings for White Balance

Monitor Settings The monitor settings are decisive for the image you see on the monitor. Set
your monitor to the appropriate color temperature, should your device allow
this. To do this, compare the image's color impression on the monitor and in
the eyepiece.
Note The color temperature setting can be altered on most monitors via the
monitor menu buttons located on the monitor. In other cases, the color
temperature can be altered via the operating system.
This is how you change the color temperature using the oper-
ating system:
1) Right click on Desktop.
" The Desktop's context menu will be opened.
2) Select the entry Properties from the context menu.
" The Display Properties dialog box will be opened.
3) Select the Settings tab.
4) Click the Advanced button.
" Another Display Properties dialog box will be opened. The appear-
ance and function of this dialog box depends on the graphics card
installed on your system and your monitor.
" Search for a tab with the name Color or Color Management.
Note The appearance of the operating elements might differ from the description
given here because the adaptation of the color temperature is not part of the
operating system, but rather is made accessible by the monitor manufac-
turer or the graphics card being used.
5) Select a color temperature whose color impression best corresponds
to that seen in the eyepiece.
" Should your system offer a slide control with the options warm and
cold, do not alter your color temperature with these.
Appendix II: How to install the camera

40
Warning Please read the following carefully before attempting to install the
camera.
Requirements
To run your DP25 you have to use the Microsoft Windows XP operating
system.
Installation sequence In order to avoid problems, you’ll have to follow the steps of the procedure
in the order given here.
Warning Install the image-analysis software first before connecting the camera
Windows 2000 to the FireWire board.
1) Check whether your computer system has a FireWire interface

onboard or a FireWire board installed.
2) Install the FireWire board provided with the camera.
3) Install the image-analysis software on your computer.
4) Install the camera and connect it to the computer system.
Requirements
What if my computer sys- If your computer system already has a FireWire interface onboard or a sepa-
tem already has a rate FireWire board installed, you do not need to install the board provided
FireWire interface? with the camera. If you are not sure whether a FireWire interface is available,
open the Windows Device Manager via Start > Settings > Control Panel >
System > Hardware > Device Manager....
An entry called ’IEEE 1394 Bus host controllers’ shows that the needed
FireWire interface is available.
General Warnings!
41
• CCD image sensors are easily damaged by static discharge. Do not
disassemble the camera housing.
• Do not attempt to touch the internals of the camera with your bare
hands.
• Do not touch cover slip (over the CCD chip) with your hand or have any
object come in contact with the glass surface. Should dust stick to the
cover slip, blow it off gently with an air blower. (For dust stuck due to
static electricity, ionized air is recommended.)
• It is best to use a LensPen to clean the cover slip. Do this only when
absolutely necessary.
• Do not expose to strong light (sunlight) for long periods.
• Exposure to high temperature or humidity will affect the characteristics
of the camera. Avoid storage or usage under such conditions.
• Do not subject the camera to too much mechanical shock.
• The FireWire cable plug is not symmetrical. You must make sure that
the cable plug is facing the same way as the camera socket when you
plug the cable into the camera jack.
DP25 • How to install the camera
• Do not disconnect the FireWire cable while the image-analysis

program is running.
The FireWire board IEEE-1394

Warning! • The components of these devices are easily damaged by static
discharge. Wear grounding wrist straps when handling them.
• Do not plug in or unplug the cards from the PCI slot while the PC is on.
• Please follow the installation manual in this booklet step by step to
avoid any incorrect connections. Incorrect connections may result in
serious damage to your PC system and/or the camera.
How to install the camera

Installation of the FireWire board IEEE-1394
When is this necessary? This procedure is necessary only if the PC does not have a FireWire inter-
face on-board hardware configuration or if it is not yet equipped with a
FireWire board.
The FireWire board
42
How to install the 1) Turn off the PC and disconnect the power supply.
FireWire board 2) Remove the cover from the PC.
3) One PCI slot is required for the FireWire board. Insert the board into the
slot.
Installation of the image-analysis software
FireWire board installed
4) Replace the cover onto the PC.

5) You are now ready to first install the image-analysis software and then
the camera.

Warning Install the image-analysis software first before connecting the camera to the
FireWire interface.
Requirements, PC The DP25 requires the image analysis program analySIS FIVE, cell^A,
cell^B, cell^D, cell^F or cell^P minimum build 1175, as well as Microsoft
Windows XP Pro with service pack 2. The image-analysis software also
supports multithreading on multiprocessors, thus making full use of the
power of the latest multi-CPU PC’s.
camera driver installation Installation of the camera driver is simply done via the image-analysis
program setup. You are only required to select DP25 as camera during
43
setup. Then you have to get Windows to use the OLYMPUS CORPORA-
TION FireWire camera driver.
Installation of the image- For installation, follow the instructions given on the monitor. You do not have
analysis software to select a digitizer in the Input Device (Digitizer) Selection dialog box.
Related topics
Installation of the camera driver, S. 48
Select Olympus as manu-

facturer and DP25 as cam-
era.
Precalibration You will be advised to carry out a precalibration of the camera when
installing the image-analysis software.
In this dialog box you can
activate the precalibration.
44
Select the TV adaptor's magnification from the list whith the same name.
Should you not be working with a remote microscope, clear the check box.
Then, mark the check box for all mounted objectives in the Magnification of
the microscope objectives list.
Note Should you use objectives with a magnification that does not appear in the
list, a manual calibration must be carried out for these objectives. Instruc-
tions on how to do this are part of the image analysis system's step by step
guide.
Note The image-analysis software can support several FireWire cameras of the
same, or different types, simultaneously. You differentiate DP25 cameras
via their serial numbers when creating a new logical input channel via Image
> Set Input... > New Channel. A camera’s serial number is located directly
below the FireWire jack on the camera housing.
The camera’s serial number
is located below the
FireWire jack.
45
How to install the camera 1) The diagram shows a typical installation of a light microscope. You
should have all required components ready at hand.
FireWire-cable
DP25
C-Mount adaptor
PC with image analysis program

and FireWire-port
FireWire-
plug
46
2) Remove the protective cap from the camera.

3) Firmly screw the TV adaptor (1/1,8" or bigger) onto the camera. This
should be done with the CCD-chip side of the camera facing down, to
minimize the chance of dust landing on its surface.
4) Install this assembly on the microscope's camera mount. Some micro-
scopes may require the installation of additional optical elements
between the camera mount and the C-mount adapter. You may also
need to align the orientation of the microscope with that of the stage,
and also match the focus of the camera image with that of the eyepiece
image. These settings can be made later, as soon as you are able to
obtain an image on the screen.
5) Plug one end of the FireWire cable into the camera connector.
" The FireWire cable plug is not symmetrical. You must make sure
that the cable plug is facing the same way as the camera socket
when you plug the cable into the camera jack. The camera chip
may be ruined otherwise.
Make sure the cable plug is

facing the right way before
you plug it into the camera
socket
6) Plug the other end into one of the small connector sockets on the
FireWire board.
" There are two or more jacks depending on the FireWire board. You
can use any of them.
7) Continue with the camera driver installation before starting the image-
analysis software.
47
Installation of the camera driver

The OLYMPUS CORPORATION FireWire camera driver is not signed and
therefore not used automatically under Windows XP. After you have
installed the image-analysis software and connected the camera to the
computer - or vice versa - you have to go through the following steps to get
Windows XP to use the OLYMPUS CORPORATION camera driver:
1) Open the Windows Device Manager via Start > My Computer > (right
mouse) > Properties > Hardware > Device Manager....
48
" The Device Manager dialog box shows the Generic 1394 Desktop
Camera entry in the Imaging devices section. The exclamation
mark indicates a device conflict.
2) Select the Generic 1394 Desktop Camera entry and click on the
Update driver button in the device manager button bar.
" The Found New Hardware Wizard dialog box will be opened.
3) Select the No, not this time option and click the Next button.
49
4) Select the Install from list or specific location (Advanced) option and
click on the Next button.
5) Select the Don’t search. I will choose the driver to install. option and
click on the Next button.
50
" The Model may already contain one ore more entries based on
previous software installations.
6) Select the entry OLYMPUS CORPORATION FireWire Camera 2.0.31
(SoftHard) or higher.
" A potentially offered driver called Olympus Soft Imaging
Solutions ... is not suitable for using with the DP25.
" If this entry is missing, click on the Cancel button. Repeat the instal-
lation of the image analysis program including the selection of
DP25 as camera.
" Do not select an entry reading Olympus Soft Imaging Solutions
FireWire Camera.
7) Click on the Next button.
" The next Hardware Update Wizard dialog box will be opened in the
background displaying the message to wait while the Wizard
installs the software.
" Additionally, the Hardware Installation dialog box will pop up in the
foreground.
8) Click on the Continue Anyway button.

" The Hardware Update Wizard dialog box which had been in the
background, will become visible.
51
Related topics
Installation of the image-analysis software, S. 43
" After having finished the installation of the OLYMPUS CORPORA-

TION camera driver, the following dialog box will be opened.
52
9) Click on the Finish button to end the hardware update wizard.

" In the Device Manager dialog box there is still the same Generic
1394 Desktop Camera entry but the exclamation mark indicating
the device conflict should have disappeared. The camera is now
ready for use.
When encountering problems with the camera...
When encountering problems with the camera...

Should you have questions or should problems arise which you cannot solve
with the help of the documentation provided, please contact your dealer.
Please clarify beforehand when, and under what circumstances, the
problem arose. It would be best if you can clearly reproduce the problem/
error. You will therefore make it easier for yourself and our customer-service
personnel to solve the problem. Proceed as follows to gain the data neces-
53
sary before you contact your dealer:
1) Please write down the exact wording of the (error) messages which
appear.
2) As an alternative, you can make "Snapshots" of these messages. You
can make a snapshot of your active window by using the key combina-
tion [Alt+Print]. A copy of the active window will then be stored to your
clipboard and can be easily attached to an E-mail using [Ctrl+V].
3) Since very large E-mails can create transmission problems both for the
sender and the receiver, do not copy "screenshots" of the entire con-
tents of the screen.
4) Should the camera basically work, but problems with image quality
arise, save an image in which the problems are clearly visible in TIF for-
mat. Insert this example image in your E-mail to our customer service.
5) Start the image-analysis software.
6) Select the ? > Info... command to open the Info dialog box.
" The Info dialog box shows you information about the software
package (Docu, Auto, Pro) of your image analysis program, the
DP25 • When encountering problems with the camera...
serial number of your image analysis program and the operating

system being used.
• The software data mentioned should be at hand when you contact
our customer service personnel.
54
WEEE DECLARATION
WEEE DECLARATION
Declaration of In accordance with European Directive 2002/96/EC on Waste Electrical and
Conformity Electronic Equipment this product manufactured by Olympus Soft Imaging
Solutions GmbH must not be disposed of as unsorted municipal waste, but
should be collected separately. The end user should refer to his local distrib-
utor for return.
55
CE Declaration of Conformity
Index
Index Exposure time, intX 10
A F
analySIS, version 4 Firmware Version 31
Automatic Gain Display Fluorescent microscopy
Input 32 Black balance 22
B G
Binning 35 Gamma Correction
Black balance 22 RGB mode 18
C I
Camera configuration 28 Image scaling, Display tab 33
Camera Control intX
Black balance 22 Calibration 9
Color Settings 18
Partial Readout 21
Sharpen filter 17
Sharpness Monitor 20 K
Camera resolution Köhler illumination, microscope 38
In input channel 30
Camera Type 31
Clipping 35 L
Color Settings 18
Gamma Correction 18 Live overlay 33
Saturation 19
Color temperature of the monitor 40
Configure Input 29 M
57
Microscope 37
Empty magnification 39
D Illumination 37
Device ID 31 Numerical Aperture 39
Device Information 30 Objective 39
Display tab Total magnification 39
Image scaling 33 Useful magnification 39
Over exposure 31 Microscope Settings 37
DLL version, F-View II III information 31 Mirror 30
DP25 Information 30 Monitor Settings 40
Duplicate Channels 29
N
E NetCam 23
Exposure time in the internet 24
Input 30 In the intranet 24
Index
Settings 26
New Channel 28 U
Useful magnification, Microscope 39
O
Online histogram 33 W
Over exposure 31 Web Server 24
Over Exposure Warning 31 White balance 16
Execute 16
P
Partial Readout 21
PC, Requirements 4
R
Requirements, PC 4
Resolution
Input 30
S
Select device 29
Serial Number 31, 54
Set Input 28
Sharpen filter 17
Activate 30
Sharpness Monitor 20
Special 16
58
T
Tab Format
Binning 35
Clipping 35
Image format 35
Tab Input
Automatic gain display 32
Exposure time 30
Info 30
Mirror 30
Resolution 30
Sharpen filter 30

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Manual

© Olympus Soft Imaging Solutions GmbH

SYSTEM REQUIREMENTS ......................................................4

WHEN CAN COLORS APPEAR TO BE FALSE? .....................6

Acquiring images with manual control 10

FURTHER OPTIONS IN THE CAMERA CONTROL ..............16

INSTALLING THE WEB SERVER ..........................................24

Setting up the camera 28

LOGICAL INPUT CHANNELS ................................................28

Appendix I: Microscope settings 37

Appendix II: How to install the camera 40

How to install the camera 42

When encountering problems with the cam-

The DP25 camera

Overview of the chapters

"Appendix II: How to install the camera" on page 40 contains detailed

2) Click the Acquire > button.

When can colors appear to be false?

When is it necessary to perform a white balance?

When can the image be too dark?

The lower part of the image

2) Click the Set ROI for White Balance button.

With the default setting of

2) Move the Exposure adjustment slide control to the right.

Calibrating Intelligent Exposure

The second calibration step consists of measuring the signals at various

Exposure Time Lock

Acquiring images with manual control

Preparing the input channel

monitor while you are making the acquisition. It is suitable for

Adjusting the exposure time

Automatic Gain Display and fixed scaling

Automatic Gain Display

2) Click the Histogram calculation on ROI to limit the image segment on

Difference between Automatic Gain Display and fixed scaling

Button Area Effect

Further options in the Camera Control

Performing a white balance with manual acquisition

If you start a live acquisition with an activated Sharpness Monitor or activate

Using the sharpness monitor

6) Click the Sharpness Monitor On/Off button to open the Sharpness

Using the Partial Readout

Using the black balance

position containing an area which is as uniformly dark as possible.

Installing the web server

You can simply adopt the default settings.

NetCam in the intranet

The transfer of images within the company network or intranet generally

NetCam in the internet

Determining the IP address

Setting up the camera

Select Device dialog box

Appendix I: Microscope settings

Adjusting the The illuminated has been optimally set.

Objective, Total Magnification and Useful Magnification

Monitor Settings for White Balance

Appendix II: How to install the camera

1) Check whether your computer system has a FireWire interface

• Do not disconnect the FireWire cable while the image-analysis

The FireWire board IEEE-1394

How to install the camera

FireWire board installed

4) Replace the cover onto the PC.