Hematoxylin/eosin counterstaining

Properly orient and embed specimen in OCT see Note 14). Quick freeze by submersing in isopentane cooled to -55 °C in the histobath. Mount specimen into the cryostat. Cut 5-pm thick sections on plus coated slides, labeled with the patients name or accession number. Fix in 10% formalin in 80% alcohol for 1 min. Rinse 10 dips in deionized water. Stain with Biomeda hematoxylin for 30 s. Rinse with deionized water. Counterstain with Eosin Y for 30 s. Dehydrate, clear in xylene, and coverslip. 

Figure 10.7 Demonstration of protein dots and tissue on single slide. This specimen was subjected to antigen retrieval prior to immunostaining, and a complete hematoxylin and eosin counterstain. 

Connterstain in hematoxylin. Other counterstains, such as Methyl Green or eosin, may be used (see Note 20). 

Counterstain as required either 1 mm m Gill s hematoxylin alone, 1% eosin alone, or hematoxylin followed by eosin will give good contrast with the black silver deposition. 

FIGURE 18.5 Light photomicrographs of microneedle-treated human skin stained for (3-galactosidase expression, (a) En face stereomicroscopy of dry-etch microneedle-treated skin with nuclear fast red counterstaining (b) en face stereomicroscopy of wet-etch microneedle-treated skin (c) hematoxylin and eosin stained 12 xm cryosection of dry-etch microneedle-treated skin. Bar =100 xm (d) hematoxylin and eosin stained 12 xm cryosection of wet-etch microneedle-treated skin. Bar = 100 xm. 

Stain 4-5 pm sections using standard hematoxylin and eosin protocols (H E) or with cresyl violet/luxol fast blue (CV/LFB). H E stains protein rich cells (eosin) and counterstains nuclei (hemotoxylin). CV/LFB also stains protein,... 

Each paraffin-embedded section is collected on microscope slides and first examined under a microscope (lOx) to ensure that it contained sufficient tumor material and to eliminate possible contaminating normal tissues. Tumor and tumor-free areas are identified within 15 pm-thick deparaffinized sections lightly counterstained with hematoxylin and microdissected by gentle scraping with sterile scalpels into 1.5 ml polypropylene vials, using a hematoxylin and eosin-stained step section from the same block [4-7]. 

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