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Journal of Histochemistry and Cytochemistry, Vol. 51, 977-980, July 2003, Copyright © 2003, The Histochemical Society, Inc.

BRIEF REPORT

HOPE Fixation of Cytospin Preparations of Human Cells for In Situ Hybridization and Immunocytochemistry

Oliver Umlanda, Artur J. Ulmera, Ekkehard Vollmerb, and Torsten Goldmannb
a Departments of Immunology and Cell Biology, Research Center Borstel, Borstel, Germany
b Clinical and Experimental Pathology, Research Center Borstel, Borstel, Germany

Correspondence to: Torsten Goldmann, Clinical and Experimental Pathology Research Center Borstel, Parkallee 3, D-23845 Borstel, Germany. E-mail: tgoldmann@fz-borstel.de

In primary or cultured cells, in situ hybridization (ISH) or immunocytochemistry (ICC) is often performed on tissue that has been fixed by paraformaldehyde or Carnoy's. Recently we reported an optimized HOPE (HEPES–glutamic acid buffer-mediated organic solvent protection effect) fixation protocol for ISH targeting mRNA in lung tissues. We have now examined whether HOPE fixation could also be used on in vitro cultured cells for targeting mRNA by ISH or proteins by ICC on cytospin preparations. Using the myeloid stem cell line KG-1a as a model system, we showed that HOPE fixation can be applied for ISH and ICC on cultured cells. HOPE can be used with cells and tissues and with a broad spectrum of immunohistocytochemical and molecular techniques.

(J Histochem Cytochem 51:977–980, 2003)

Key Words: HOPE fixation, paraformaldehyde, Carnoy's solution, cultured cells, ISH, ICC, mRNA, cytokines, cytospins


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