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Understanding Estrous Cycle Stages in Rats Using Vaginal Smear Techniques

This guide outlines the procedure for assessing the estrous cycle in rats through vaginal smears. It details the characteristics of each estrous phase, including Diestrous, Proestrous, Estrous, and Metestrous, highlighting the presence of epithelial cells and white blood cells. A step-by-step protocol for obtaining samples, staining them, and analyzing the results under a microscope is provided to help researchers accurately determine the estrous phase and maintain proper records for future reference.

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Understanding Estrous Cycle Stages in Rats Using Vaginal Smear Techniques

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Presentation Transcript

  1. Diestrous Phase Stringy mucous, white blood cells (not many in this photo), nucleated epithelial cells

  2. Proestrous Mostly nucleated epithelial cells

  3. Early Estrous Relatively even distribution of nucleated epithelial cells and cornified epithelial cells

  4. Estrous Mostly cornified epithelial cells

  5. Metestrous Many WBC, some nucleated and cornified epithelial cells

  6. Vaginal smear procedure: • Use clean pipet tip and inject about 25 microliters of 0.9% saline into the vagina of the rat. • With same tip, obtain a sample of the vaginal lavage and transfer to a clean slide (label with initials and the date). • Let dry, then stain for approximately 30-45 seconds in Wright stain (place a drop or two of stain on your slide). • Rinse slide briefly in water, then observe smear under microscope. Record description of smear and determine estrous phase. Retain slide for future reference if needed (store in labeled box in animal room).

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