Samples were fixed with 2.5% glutaraldehyde, 2% paraformaldehyde in 0.1 M sodium cacodylate buffer, postfixed with 1% osmium tetroxide followed by 2% uranyl acetate, dehydrated through a graded series of ethanol and embedded in LX112 resin (LADD Research Industries, Burlington VT). Ultrathin sections were cut on a Leica Ultracut UC7, stained with uranyl acetate followed by lead citrate and viewed on a JEOL 1400 Plus transmission electron microscope at 120kV.
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Electron Microscopy Sample Preparation
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Corresponding Organization : Montefiore Medical Center
Other organizations : Royal Veterinary College, Aristotle University of Thessaloniki
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Variable analysis
independent variables
- Fixation of samples with 2.5% glutaraldehyde, 2% paraformaldehyde in 0.1 M sodium cacodylate buffer
- Postfixation with 1% osmium tetroxide
- Dehydration through a graded series of ethanol
- Embedding in LX112 resin (LADD Research Industries, Burlington VT)
- Ultrastructural features of the samples observed under transmission electron microscope
- Fixation time
- Osmium tetroxide concentration
- Dehydration steps
- Resin composition
- Sectioning and staining procedures
- Electron microscope settings (120kV)
- No positive or negative controls were explicitly mentioned in the protocol.
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