I am working with E. Coli biofilm samples. Our method use a paraformaldehyde fixation step in order to preserve biofilm structures before microscopy observation. However, we have saw that there was a loss of biofilm (a degradation) if this step is used compared to if it is not used. I want to know if this step could be responsable for the destroy of biofilm observed.
Thank you so much!
Plz see https://aem.asm.org/content/aem/63/4/1570.full.pdf
Carlota hello,
For additional information, have a look here:
www.ncbi.nlm.nih.gov/pmc/articles/PMC3181414/
www.ncbi.nlm.nih.gov/pmc/articles/PMC4776306/
I hope this helps :)
Hi Carlota,
Generally, imagery techniques (scanning electron microscopy (SEM), transmission electron microscopy (TEM), and focused ion beam nanotomography (FIB-nt)) involve the desiccation of biofilm samples, e.g., freeze-drying or chemical dehydration. Confocal laser scanning microscopy (CLSM) allows imaging biofilm in its hydrated form. However, it does not reveal differentiation between biofilm components such as inorganic particles and pore water. great results can be achieved with X-ray microcomputed tomography (μ-CT) that enables the high-resolution illustration of morphology within a hydrated biofilm matrix but not everyone has access to this technology.
I found that the best results during the preparation of biofilm samples for SEM imaging gives the following fixative solution: 2 % glutaraldehyde and 2 % paraformaldehyde in 0.1M HEPES buffer, e.g., 100ml of the ready to use solution is made up by adding 10ml of 20 % paraformaldehyde, 8ml of 25 % glutaraldehyde, 50ml of 0.2M HEPES buffer and 32ml of sterile cell grade water to 100 mL graduated glass bottle and gently mixed.
Here is a link to one of my SEM micrographs prepared using this solution:
https://en.wikipedia.org/wiki/Biofilm#/media/File:Mixed-culture_biofilm.jpg
Please feel free to contact me if you need any further information.
Kind regards,
Chris
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