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  1. This procedure establishes a consistent process for preparing H&E slides from frozen or FFPE tissue samples using the automated H&E stainer and coverslip instruments. This

  2. Hematoxylin and Eosin Stain For Frozen Section (H&E) PROCEDURE: 1. Rinse slides in 1X PBS for . 3 minutes. 2. Rinse in a gentle stream of . tap water. until OCT is washed off for . 3 minutes. 3. Stain in . Mayers Hematoxylin. for . 30 seconds. 4. Wash in a gentle stream of . tap water. for . 1 minute. 5. Blue nuclei in . 1X PBS. for . 20 ...

  3. HEMATOXYLIN & EOSIN (H & E) STAIN PROTOCOL PRINCIPLE: This protocol is applied in the routine staining of cationic and anionic tissue components in tissue sections. This is the standard reference stain used in the study of histochemical tissue pathology. SPECIMEN REQUIRED: Snap frozen human striated muscle. (Use the 2-methylbutane freezing method)

  4. Use frozen section to Provide rapid gross or microscopic diagnosis to identify an unknown pathologic process, identify extent of disease / evaluate margins, identify metastases or simply identify a tissue Process tissue to provide appropriate and accurate diagnosis, prognosis and to adhere to research and special study protocols

  5. Protocol for H&E staining: While sections are in water, skim surface of hematoxalin with a Kimwipe to remove oxidized particles. Blot excess water from slide holder before going into hematoxalin. Coverslip slides using Permount (xylene based).

  6. Before H&E staining, frozen tissue sections should be fixed in precooled acetone for 60 seconds (acetone kept in freezer at –18 °C overnight prior to use). Slides should be stained immediately after fixation to avoid drying out.

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