Mount the dried samples on stubs containing carbon adhesives
Platinum / Palladium coating: 6 nm
Human Blood Cells Protocol
Application Note for Leica EM CPD300 - Life Science Research
Species: Human (Homo sapiens)
Critical point drying of human blood with subsequent platinum / palladium coating and SEM analysis
Samples were inserted into the 12 mm cover slip holder
Place 12 mm dia cover slip poly-L-lysine coated in a 12-wells cell culture plate. Add 1 ml 0.85% NaCl in each well to submerge each cover slip.
Pipette gently 50 μl blood on each glass cover slip leave for 5 min at 25°C.
Add 200 μl 0.2 M CaCl2 on top of the blood cells to activate the platelets and leave for 10 min.
Fixation and Dehydration:
Add gently 1 ml of 4% Paraformaldehyde, 0.4% Glutaraldehyde in 0.2 M Sodium Cacodylate Buffer, pH 7.2, on top of the blood cells and leave at least for 10 min. at RT.
|Distilled Water||3x10 min.|
|1% aqueous OsO4, 4°C||16 h|
|Distilled water||3x10 min.|
|Ethanol series: 30%, 50%, 70%, 80%, 90%, 96%, 100%||1x10 min.|
|Acetone series: 30%, 50%, 100%||1x10 min.|
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