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Hi,
I am looking for any advice or recommendations for sample preparation in the case of in situ imaging of protein (antibody) adsorption on mica in fluid using tapping mode. I am using Nanoscope IIIa and MTFML liquid cell.
I have problems with right sample injection procedure into the fluid cell, because I have constantly problems with fluid leaking or I don't observe any adsorbed sample on the mica. I start with DI water measurement, then retract the cantilever and inject the sample into the cell and finally wait for 10-15 min. But it seems that I'am doing something wrong.
I am performing this kind of measurement to prepare procedure for subsequent imaging of protein crystallization process.
If anybody has experiences with protein adsorption or crystallization imaging, could you share them?
Thank you,
Dejan
Hi, Dejan
What type of o-ring are you using?
Usually to get a good seal. I engage in air first. Then I inject the fluid through the tubing to make sure there is no leak. I like to use 1 ml syringe for my in-port and the silicon tubing with stop clamp on the out let. To avaid air bubble during buffer exchange when changing the syringe, I wet the in-port. once you have a good seal at the begining, the fluid cell should stay seal.
http://www.cma.fcen.uba.ar/files/prepmuestras.pdf
Here is a link to general afm prep for bio sample.
Depends how well your antibody stick to mica, you may need to silanize the substrate.
-james