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Freeze-drying Allows Double Nonradioactive ISH and Antigenic Labeling

2000; SAGE Publishing; Volume: 48; Issue: 4 Linguagem: Inglês

10.1177/002215540004800408

1551-5044

Huguette Louis, Julie Lavie, Patrick Lacolley, Danièle Daret, Jacques Bonnet, Jean‐Marie Daniel Lamazière,

Viral Infectious Diseases and Gene Expression in Insects

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.