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Sample preparation from paraffin-embedded tissue specimens for laser scanning cytometric DNA analysis by Tomomi Kamada; Kohsuke Sasaki; Tatsuo Tsuji; Takeshi Todoroki; Masahiro Takahashi; Akira Kurose is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Sample preparation from paraffin-embedded tissue specimens for laser scanning cytometric DNA analysis about?

We have developed a simple, rapid method for isolating cells from a block of formalin-fixed, paraffin-embedded tissue specimen for laser scanning cytometric (LSC) DNA analysis by using a grater. The scraping-like tissue samples were obtained by grating a paraffin-embedded tissue block. The grated samples were collected, put into a small plastic tube, and deparaffinized with xylene. Subsequently, the samples were immersed in 100% ethanol to remove the xylene. After using a syringe with a 26-gauge needle and filtering through 40-mm nylon mesh, the cells suspended in ethanol were dropped directly onto a glass slide. As a result, isolated cells adhered tightly to the glass slide. The slides mounted with isolated cells were treated with 0.1% pepsin in 0.1 N HCl for 1 h at 37°C and then 0.1% RNase for 10 min at room temperature. The slides were dipped in propidium iodide (25 mg/ml) to stain DNA and sealed with nail varnish. The coefficients of variation for histograms were small enough to detect an aneuploid peak close to the diploid peak.

Who reads Sample preparation from paraffin-embedded tissue specimens for laser scanning cytometric DNA analysis?

Author

Tomomi Kamada; Kohsuke Sasaki; Tatsuo Tsuji; Takeshi Todoroki; Masahiro Takahashi; Akira Kurose

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; John Wiley & Sons Ltd.; Wiley (ISSN 0196-4763)

Published

1997

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)