Good morning,
I would like to ask for help in analyzing the SEM image. The photo shows HMEC cells growing on the surface of E-UV modified PVDF. Preparation of the sample for SEM analysis: incubation in a 4% solution of paraformaldehyde/PBS, dehydration in an ethanol/water solutions (50, 60, 70, 80, 90, 100%; 5 minutes each). I will be grateful for the hint what the bright objects can be? They are visible mainly on the surface of cells.
Hello,
So bright and with sometime geometrical forms, I will say salt from PBS.
Thank you, Nikolas, for your answer. The samples were air-dried. I also thought about PBS crystals, but it seems to me that after rinsing several times in a water-ethanol solutions, PBS should be rinsed off (the material is flat, without pores). In addition, crystals are only present on the surface of the cells, they are absent on the material. A PFA residue comes to my mind, maybe it is worth filtering the PFA solution?
You should wash with water before going with 50% ethanol.
Air drying gives poor surface preservation.
For drying, if you don't have a critical point dryer (a machine drying the sample by changing the phase of CO2 from liquid to gaz beyond its critical point), I suggest to use the hexamethyldisilazane (HMDS).
After 100% ethanol, you go for 2 bathes of 100% HMDS (under the hood), then you remove the HMDS and let it dry.
Beata Butruk-Raszeja Nicolas Barois can you tell me why my mutant not equally replicate ? In SEM experiment
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