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STAINING PROTOCOL
1) After last PBS wash, rinse in tap H20.
2) Filtered hematoxylin: 15 min cytospins 15 min tissue
3) Rinse in tap H20 3 to 4 times until clear, then keep in H20 another 10 minutes until staining has "blued".
4) (IF COUNTERSTAINING WITH EOSIN/PHLOXINE B) 70 -t ETOH 2 min. Eosin/phloxine B (filtered) 3 minutes (cytospin or tissue). Eosin can be used 2 to 3 times, then discard.
5) Graded alcohol series 2 min each
70 X3
95 X2
100 X2
1:1 ETOH/XYLENES Xl
XYLENES Xl
6) Coverslip. Mount. Dry o/n.
NB: Both hematoxylin and eosin/phloxine B were obtained from American Histolabs.
DEPARAFFINIZATION PROTOCOL
1) Approximately 5 min. in xylenes X2 to remove paraffin
2) Hydration series, 2 min. each
100 ETOH X2
95 ETOH X2
70 ETOH X2 PBS X3
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