If we have scanning electron microscope with magnification up to 100000x, and the samples holder can freeze the samples, we see only the crystals. We can't see the cells, or the bacteria, or the viruses. What can we do, or how can we prepare the sample for better results?
We have phenom Pro SEM which is
Key Specifications
Magnification range 20 - 100,000x
Long-lifetime high-brightness source (CeB6)
5 kV and 10 kV acceleration voltages for outstanding high-resolution SEM images
Sample loading in less than 30 seconds
Full color Navigation Camera 20 – 120x magnification
Ready to run all Pro Suite software applications
Very low energy consumption
http://www.phenom-world.com/electron-microscope/phenom-pro-series/phenom-pro#description
Your microscope has a Peltier stage, which does not give Cryo SEM capabilities. There were some papers which described utilization of Peltier stage freezing for observation of plants (some plants can withstand rough freezing ), but after considerable efforts and by highly trained operators. So, in general, your microscope is not suited for observation frozen biological specimens. I advise you to forget about this cooling stage and prepare your specimens in usual way: fixation, dehydration, critical point drying (or HMDS) and sputter coating. You can find a lot of protocols in books and on the Web.
Dear Ahmed,
Please see the following publication for fixation of mammalian cells for SEM:
https://www.researchgate.net/publication/8477611_Porosity_variation_in_hydroxyapatite_and_osteoblast_morphology_a_scanning_electron_microscopy_study?ev=prf_pub
Please note that used Osmium tetroxide is a hazardous substance and should be discarded as hazardous waste. Please check the appropriate procedure for your department.
Decontamination of OSO4 can be found in the following link as well: http://naples.cc.sunysb.edu/Admin/HRSForms.nsf/pub/EHSD0434/$FILE/EHSD0434.pdf
Best Wishes
Basil
Article Porosity Variation in Hydroxyapatite and Osteoblast Morphology
How to prepare dry mycelium for SEM analysis ??
I want to SEM (Scanning Electron Microscop ) analyzes on samples that had been dried mycelium with a freeze drier Thanks
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