I am a PhD student in a single molecule biophysics lab. My kinesin decorated beads are sticking to the surface at times, even with casein blocking. (I have to wash out casein ensure good image quality)
Currently using repel silane es to coat borosilicate coverslips, and then blocking with pluronic for 15 mins, and then adding additional casein to help block. This works, but I think I can silanate the glass using a less expensive silation alternative.
I am looking at n-OCTADECYLDIMETHYLCHLOROSILANE, but I would like to know if a trichlorosilane is better to ensure a more robust coating.
Thank you,
Jared
I think the n-OCTADECYLDIMETHYLCHLOROSILANE is one of the best and will probably give you a contact angle of around 110 degrees or so. Initially I though dichlordimethyl silane as that is cheaper, but your thoughts would make it more hydrophobic.
If you want to increase hydrophobicity even further could think about using a fluorinated silane such as tridecafluorooctyl triethoxysilane or Trichloro(1H,1H,2H,2H-perfluorooctyl)silane. These are more expensive.
I would suggest not much difference between borosilcate glass and silica (quartz) I would suggest
Paul
Hi Jared, personally, I would go for the trimethoxy- (or triethoxy) silanes because they will be bonded to glass with 3 Si-O- bonds.
The 3 bonds will be harder to displace than single Si-O- bonds in the presence of -OH compounds like pluronic and casein.
Plus, they do not generate HCL upon binding to glass.
Hi. Reading your question, it reminded me of this recent methods paper by the TJ Ha lab: http://www.nature.com/nmeth/journal/v11/n12/full/nmeth.3143.html
Maybe it is of use?
Good luck!
Willem
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