Tissue Fixation and Single-Cell Isolation
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Corresponding Organization : Vanderbilt University
Variable analysis
- Tissue fixation method (10% formalin for 24 to 72 hours)
- Tissue embedding method (paraffin)
- Tissue sectioning thickness (5 μm)
- Tissue staining methods (hematoxylin and eosin, trichrome, or appropriate antibodies for immunohistochemistry)
- Tissue digestion method (enzymatic cocktail [collagenase I/dispase II (1 μg/ml) tissue or Miltenyi Multi Tissue Dissociation Kit] using a gentleMACS Octo Dissociator)
- Microscopic examination of tissue sections under light microscopy
- Tissue storage buffer (MACS Tissue Storage Solution, Miltenyi Inc.)
- Sterile filtration process (sterile gauze, 100- and 40-μm sterile filters)
- Freezing of cell suspensions (90% fetal bovine serum/10% dimethyl sulfoxide, -80°C)
- Positive control: Not specified
- Negative control: Not specified
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