I'm hoping to get started learning how to work with the optical-tweezers apparatus soon. One of the things I want to test is adhesion of H. influenzae cells to silane-coated coverslips, as this will eliminate the need to restrain the cells by suction from a micropipette. The RA has already found that cells don't stick to uncoated coverslips, nor to ones coated with BSA. I had been hoping that I could just order them from Sigma or somewhere, but now it looks like preparing the coverslips might be the first of many very fussy procedures.
A google search didn't find anyone selling them, but it found lots of protocols for making them. They all involve precleaning the coverslips by boiling in strong acid or soaking in strong base or scrubbing with bathtub cleanser (Bon Ami was used), and then dipping or soaking them in solutions containing 3-aminopropyltriethoxy-silane/epoxy-silae/TMA silane/trichloro-octenyl-silane/methacryloxypropyltrimethoxysilane, and then air-drying or baking or ??? and storing in a dessicator or under unspecified conditions. I have no idea what kind of silane I should use or how to prepare the coverslips, so I've emailed the author of the Bacillus subtilis paper that did tweezers analysis, to ask what she did. I've also emailed my physicist collaborator (whose apparatus I'll be using) to ask if she has experience with this.
I think I'll also email around our institute, to find out if anyone here is routinely using silane-treated cover slips. Maybe they can (1) give me one to test, and (2) show me how to make them.
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in The Biology Files
Not your typical science blog, but an 'open science' research blog. Watch me fumbling my way towards understanding how and why bacteria take up DNA, and getting distracted by other cool questions.
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