Frozen Section Notes PDF

FROZEN SECTION Prepared by: Giane Paola T. Tanjuaquio, RMT  FROZEN SECTIONS  Normally utilized when a rapid diagnosis of the tissue in question is required, and is especially recommended when lipids and nervous tissue elements are to be demonstrated  Cryostta: a cold chamber kep...

FROZEN SECTION Prepared by: Giane Paola T. Tanjuaquio, RMT  FROZEN SECTIONS  Normally utilized when a rapid diagnosis of the tissue in question is required, and is especially recommended when lipids and nervous tissue elements are to be demonstrated  Cryostta: a cold chamber kept at an atmospheric temperature of -10C to -20C  Very thin slices, around 10-15 microns in thickness are cut from a fresh tissue frozen on a Cryostat, a cold chamber kept at an atmospheric temperature of -10°C to -20°C  FROZEN SECTIONS ARE COMMONLY USED FOR: 1. Rapid pathologic diagnosis during surgery 2. Diagnostic and research enzyme histochemistry 3. Diagnostic and research demonstrations of soluble such as lipid and carbohydrates 4. Immunofluorescence and immunohistochemical staining 5. Some specialized silver stains, particularly in neuropathology  COMMONLY USED FREEZING AGENTS: 1. Liquid nitrogen 2. Isopentane cooled by liquid nitrogen 3. Carbon dioxide gas 4. Aerosol sprays  COMMON SPECIMEN SUBJECTED TO FROZEN SECTION PROCESSING 1. Lymph nodes (Sentinel lymph node biopsy)  Submitted fresh, dampened with saline  Sentinel node biopsy is recommended for people with certain types of cancer (breast and melanoma) in order to determine whether the cancer cells have migrated into the lymphatic system.  Remove fats before freezing  For DNA and RNA extraction  Snap freezing  COMMON SPECIMEN SUBJECTED TO FROZEN SECTION PROCESSING 2. Fatty specimens such as breast and lymph nodes (-25 to - 35°C) 3. Colon (-15 to -25°C) 4. Skin lesions (-15 to -25°C)  COLD KNIFE PROCEDURE 1932, Schultz-Brauns Knife -40 to 60t Tissue: -10°C Environment 0-10 °C  FREEZE DRYING Rapid freezing (Quenching) at -160 °C then removal of ice water molecules (Desiccation) in a vacuum at-40 °C (sublimation)  FREEZE SUBSTITUTION Same as Freeze Drying except that tissues are fixed in Rossman's formula or in 1% acetone and dehydrated in absolute alcohol.  OCT (Optimal Cutting Temperature) compound  FREEZING MICROTOME  CRYOSTAT  Set according to the type of tissue being examine.  Average temperature: -20°C  All frozen section tools are also cooled inside the cryostat chamber.  STAINS 1. H&E (progressive) Method 2. Polychrome Methylene Blue Method 3. Alcoholic Pinacyanol Method 4. Thionine 5. Rapid Giemsa staining method 6. Rapid Eyedropper staining method: toluidine blue or thionine  COVERSLIPPING Mounting fluid – Glycerol – Gum Arabic and other aqueous media – Commercially available: Clearcol, Viscol, Abopon, Paragon