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Hi,
I want to get high resolution images of antibody sample on mica in fluid (buffer solution). I have Nanoscope IIIa and MTFML liquid cell.
I tried with small cantilever (38 um length) from Olympus: BioLever Mini (BL-AC40TS) with ~0.1N/m and ~30kHz in water. For now I have problems with high noise in obtained images and images have large spikes. Now, I don9t now if it is better to increase/decrease scanning rate or gains.
Do you have any experience with these cantilevers for this particular application or any other suggestions?
Thank you for your consideration of this matter,
Dejan
Hey Dejan,
I am trying the same thing (Multimode with MTFML and BL-AC40TS) and I also have the same problems... so if you find any solutions, please let us know here.
Did you also try to use SNL cantilevers? I'm looking at small protein filaments in liquid and these levers did already a quite good job.
Cheers, Raphael
Hi Raphael,
sure, if I will find solutions I will post them.
I didn't try SNL cantilevers yet, but I will.
BTW, how did you immobilize your protein sample on the mica?
Regards,
SNL or DNPS can me good choice. You can try the 0.24 or 0.35 N/m levers.
The key to success is to image under light tapping conditions (ratio setpoint around 90%). See the related publications by Alvaro San Paulo and Ricardo Garcia (1999 and 2001 if I remember correctly). Of course I assume you know the golden rule: find a peak around 8-12 KHz...
Good luck,
Alex.
In my case, I do not use any cross-linker at the moment. Freshly cleaved mica works fine... but maybe you can use AP-mica (APTES) or even glutaraldehyde.
@Alex
I know the article that you suggested. I've already got decent images in air using their approach (but with slightly softer cantilever 26N/m). but now I want to obtain high resolution images in fluid (maybe also force volume). And I am searching for the right cantilever.
@Raphael
I don't use any cross-linker either. But, I am planing to use buffers with same sign of charge like mica and I will probably have to use AP-mica.