Special stains in

Histotechnology:
Part II
Dr CN Srinivas
Director- Dept of Lab Medicine
MIOT International
Chennai

Dr C N Srinivas
MD,DNB(Path),IFCAP

Objectives

Perls Prussian Blue

GMS
Fite Farco stain
Quality Management in HP and
Cytology

Perls
Prussian blue
Principle
Dilute mineral acid hydrolysis releases ferric ions from
protein bound tissue deposits, which, in the presence of
ferrocyanide ions, is precipitated as the highly colored
and highly water-insoluble complex, potassium ferric
ferrocyanide, Prussian blue.
FeCl3 + K4Fe(CN)6 = KFeFe(CN)6 + 3KCl

Ferrous ions do not produce a coloured reaction product,

thus are excluded from the visualization.
Tissue deposits containing ferric ions are invariably
haemosiderin.

utility
Hemosiderin demonstration in Liver
biopsy
Urine deposit
Bone marrow for iron stores
Bone marrow for MDS classification

Sideroblasts

Tips and Tricks

Use Iron free reagents
Water from Taps should not be used
Make sure fresh solutions are made

Gomori Methamine Silver

Method
1. Oxidation of the tissue:
Chromic Acid is used.
Oxidation and in addition, releases
aldehyde groups to further
breakdown products that will NOT
combine with final Silver solutions.
(Further Oxidation)Thus Collagen fibres and BM
are supressed to take the silver stain

2. Removal of Traces of Chromic acid by

Sodium bisulfite
3. Water wash and treat with Alkaline
Silver solution
4. Silver solution reacts with oxidized
products which reduces silver nitrate to
metallic silver.
5.Methenamine : Alkaline property
6. Sodium borate: Buffer

 7. Sodium thiosulfate fix silver

reaction and remove underutilized
silver.

Fungus
Pneumocystitis Carinii
Basement membrane of Glomerulus
Bone marrow fibrosis

Fite Farco stain

Demonstrate of Lepra bacilli
PhenylMethane
dye( Pararosaline,Rosaline or new
fuchsin) in a phenol solution is used.
Phenol : enhances staining with Fuchsin
dye and dissolves the same during
stain preparation
Alcohol : Carbol-fuchsin to enhance
staining and dissolve the dye

 When carbol Fuchsin is added all the

bacteria are stained as red
Acid Alcohol decoloriser is added.
The lepra bacilli holds the Carbol
Fuchsin dye rest becomes bleached.

Brain to Brain loop

ISO 15189:2012

Quality Management in
Anatomic Pathology/Cytology
Pre analytical- Fixation, Grossing and TRF
Analytical- (Quality Control)(5M1E)
Man, Machine,
Method,Material,Money,
Environment
Post analyticalInterpretation of the stain,
Pathologist

Tips to improve stains

Personnel training
Quality control blocks
Fresh viable reagents
Deionised water
Coplin jars to be neat and clean
Labelling
Quality mountants

Tips to improve stains

Competence
Good Microscope
Correlation with HE and special stains