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Journal of Histochemistry and Cytochemistry, Vol. 48, 499-508, April 2000, Copyright © 2000, The Histochemical Society, Inc.

ARTICLE

Freeze-drying Allows Double Nonradioactive ISH and Antigenic Labeling

Huguette Louisa, Julie Laviea, Patrick Lacolleyb, Danièle Dareta, Jacques Bonneta, and Jean-Marie Daniel Lamazièrea
a U441 INSERM, Pessac, France
b U337 INSERM, Paris, France

Correspondence to: Jean-Marie Daniel Lamazière, U441 INSERM, Avenue du Haut Lévêque, 33600 Pessac, France. E-mail: Jean-Marie.D-Lamaziere@bordeaux.inserm.fr

Because tissue freeze-drying is an excellent way to preserve antigenic conformation, we have tested the feasibility of this technique to reveal nonradioactive in situ hybridization (ISH) of tissue mRNA. We have compared mRNA detection after different methods of tissue preservation, freeze-drying, cryosectioning, and formaldehyde or methanol fixation. Our results show that nonradioactive ISH is more sensitive for tissues preserved by freeze-drying than for other tissue preparations. We have demonstrated that freeze-drying allows combination of ISH and immunohistochemistry for simultaneous detection of mRNA and antigen because with this technique of tissue preservation ISH does not affect the sensitivity or the amount of the detected antigens. This work underscores the fact that tissue freeze-drying is an easy, convenient, and reliable technique for both ISH and immunohistochemistry and achieves excellent structural conditions for nonradioactive detection. (J Histochem Cytochem 48:499–507, 2000)

Key Words: in situ hybridization, freeze-drying, nonradioactive detection, double detection


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