Silanizing Glassware

Glassware is silanized (siliconized) to prevent adsorption of solute to the glass surface or to increase its hydrophobicity. This is particularly important when dealing with low concentrations of particularly sticky solutes such as single-stranded nucleic acids or proteins or cells. Materials Chlorotrimethylsilane or dichlorodimethylsilane Vacuum pump Desiccator, equipped with a valve 1. In a fume hood, place glassware or equipment to be silanized into desiccator along with a beaker containing 1 to 3 ml of chlorotrimethylsilane or dichlorodimethylsilane.
CAUTION: Chlorotrimethylsilane and dichlorodimethylsilane vapors are toxic and highly flammable.

APPENDIX 3E
BASIC PROTOCOL

2. Connect desiccator to vacuum pump until silane starts to boil and close connection to pump (maintaining vacuum in desiccator). Leave the desiccator evacuated and closed until liquid silane is gone (1 to 3 hr).
During the incubation the silane will evaporate, be deposited on the surface of the glassware, and polymerize. Do not leave the desiccator attached to the vacuum pump. This will suck away the silane, minimizing deposition and damaging the pump.

3. Open desiccator in a fume hood, and leave open for several minutes to disperse silane vapors. 4. If desired, bake or autoclave the glassware or apparatus.
Autoclaving or rinsing with water removes the reactive chlorosilane end of the dimethylsiloxane polymer generated by dichlorodimethylsilane.

COMMENTARY
Untreated glass contains silicate and silanol groups that can act as ion-exchange and nucleophilic centers. To mask these groups and decrease the hydrophilicity of the surface, various reactive silanes are frequently used to coat the glass surface. The same or related chemistries can be used to introduce functional groups, including large molecules, onto the glass surface. On contact with a silanol (SiOH) group, chlorosilanes react to give HCl and a siloxane Si-O-Si linkage. For most applications silanizing or siliconizing a piece of glassware or equipment means introducing onto the glass surface either a short polymer of dimethylsiloxane
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using dichlorodimethylsilane, or a trimethylsiloxane cap

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using chlorotrimethylsilane. Polydimethylsiloxane is silicone oil; the use of dichloromethylsilane essentially gives a light coating of oil to the glassware. The polymerization of dichlorodimethylsilane is mediated by residual water adsorbed to the glass surface; chlorosilanes also react with water and alcohols to give HCl and silanol or alkoxysilane, respectively. Thus, in the polymerization reaction, the residual water reacts with the chloride endgroup to give HCl and a silanol, which then reacts with more dichlorodimethylsilane and more water to give
Commonly Used Techniques

Contributed by Brian Seed

Current Protocols in Cell Biology (2000) A.3E.1-A.3E.2 Copyright 2000 by John Wiley & Sons, Inc.

A.3E.1
Supplement 8

polymers. Chlorotrimethylsilane just gives a trimethyl cap to the silanol or silicate group. Items too large to fit in a desiccator can be silanized by briefly rinsing with or soaking in a solution of approximately 5% dichlorodimethylsilane in various volatile organic solvents such as chloroform or heptane. The organic solvent is removed by evaporation, depositing the dichlorodimethylsilane on the surface. This approach is particularly useful for

treating glass plates for denaturing polyacrylamide sequencing gels. CAUTION: If a flammable solvent is used, do not bake the glassware until the solvent is completely evaporated.

Contributed by Brian Seed Massachusetts General Hospital and Harvard Medical School Boston, Massachusetts

Silanizing Glassware

A.3E.2
Supplement 8 Current Protocols in Cell Biology