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Microtoming Tips
1. Tissues are microtomed at 7 m thick. 2. The number of sections are selected depending on size of tissue (i.e. gonad: one in every ten sections, thyroid: one in every five sections). 3. The sliced paraffin block becomes a fragile ribbon made of wax, as long as preferred (refer to #2), and it is gently placed into warm water bath. 4. While in the water bath the tissues can be separated with the use of a thin dissecting poker, which has been heated with an ethanol lamp. 5. Once the tissue is separated, the section is placed on an albumen slide and placed on a hotplate (you may prefer to check the sectioning quality using a dissecting
microscope to keep a closer eye on the quality prior to staining, look for either dull knife marks or desired tissues). 6. Sections should be placed chronologically in a row down the center of the slide (to facilitate viewing under a microscope)) and left on the hotplate (on low) over night.
Coating Slides 1. Slides are individually laid out on a slide warmer at a low setting (one box of slides can fit on the larger C.S.&E slider warmer). 2. A thin layer of albumin is painted onto each slide using a small clean paintbrush. 3. Slides are allowed to dry on the warming plate overnight at a low setting. 4. Albumin coated slides are stored at room temperature in the original packaging until needed.
17. Permount - add 4 small drops/slide and then carefully place cover slip before the permount dries.
Eosin (pink)- binds to eosinophilic tissue: cytoplasm, muscle fibers(red); connective tissue
fibers; secretory granules; erythrocytes(red); fiberous tissue(pale pink); and protein precipitate.
Hematoxylin (purple) -binds to basophilic tissues: nucleus (proteins and nucleic acids); RNA
and proteins in cytoplasm; mucus and mucoproteins; and cells with large amounts of ribosomal material on smooth ER.