LSPs and CPPs were fixed in 4% PFA for 30 min, cytospun onto glass slide, washed with PBS, fixed with 2% glutaraldehyde (GTA) for 1 h, washed extensively with 0.1 mol/L sodium cacodylate buffer (Electron Microscopy Sciences, Hatfield, PA), dehydrated in ethanol series and dried at RT, sputter coated using a high-resolution sputter coater (Ted Pella, Inc., Redding, CA) with a thin film of gold at 13.3 Pa and 45 mA for 90 s. The images were collected with Hitachi S4700 microscope (28 (link)). To estimate damaged PLTs in CPPs, 7 large-field images were collected, and percentages of damaged PLTs with rough, porous membranes and grape-like structures were counted.
Protocol detail
Scanning Electron Microscopy of Platelets
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Buffer
Cacodylate
Cpps
Dehydrated ethanol
Electron microscopy
Glutaraldehyde
Gold
Grape
Membranes
Microscope
Sodium
Corresponding Organization : United States Food and Drug Administration
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Variable analysis
independent variables
- Fixation with 4% PFA for 30 min
- Fixation with 2% glutaraldehyde (GTA) for 1 h
- Percentage of damaged PLTs with rough, porous membranes and grape-like structures
- Cytospinning onto glass slide
- Washing with PBS
- Dehydration in ethanol series
- Drying at room temperature
- Sputter coating with a thin film of gold at 13.3 Pa and 45 mA for 90 s
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