Hello,
I'm fixating bacterial biofilms on plastic surfaces for SEM imaging and I don't have the facilities for critical point drying, nor access to Hexamethyldisilizane (HMDS) as a drying agent.
So I wonder, what are the properties that a chemical drying agent should bring for preserving bacterial morphology? Is there any "quick and dirty" method I could use instead?
Best,
Joana
The HMDS method has be tested and even less expensive especially comparing it with the CPD method. I think you should just use the method as it is very reliable. See the procedure here
Transfer your sample from 100% ethanol into a 1:2 solution of HMDS: 100% ethanol and leave for 20 minutes
2. Transfer your sample to a fresh solution of 2:1 HMDS:ethanol for 20 minutes
3. Now transfer the sample into 100% HMDS for 20 minutes, and repeat this step
4. When you sample is submerged in the final 100% HMDS solution leave covered or capped loosely in a fume hood overnight
5. All HMDS steps need to be carried out in the fume hood wearing the necessary personal protection gear as it is highly toxic
6. The HMDS will evaporate, leaving your sample ready for sputter coating and imaging in the SEM.
Best wishes....
Dear,
I would recommend to use glass cover slides for the biofilm formation. Put your slides after the biofilm growth in a 6 wells tissue culture plate and soak the slide with 5% gluteraldehyde for 16-24 hrs in +4c. Next day discard the fixative and let it air dry from 72-96 hrs. Then coat your sample and examine it by the SEM.
Additional point for Omar's answer.
Insted of air drying, you can keep our sample in a desiccator overnight. We used that drying method for our biofilm study. (biofilms produced on glass cover slips)
Me dio excelentes resultados con nematodos, pero pesimo en hongos filamnetosos
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