PMID: 7543364Feb 1, 1995Paper

Bisbenzimide: a fluorescent counterstain for tissue autoradiography

Histochemistry and Cell Biology
S A Schnell, M W Wessendorf

Abstract

Interpretation of the data from experiments using autoradiography (e.g. using in situ hybridization histochemistry, receptor binding, neuronal tract-tracing etc.) is aided when the autoradiographic grains can be seen in the context of cellular boundaries. Studies making use of autoradiography in the central nervous system have sometimes used tinctorial stains, such as cresyl violet, as counterstains to visualize the labeling. Tinctorial stains are excellent Nissl stains however, under bright-field illumination such dyes tend to obscure autoradiographic grains. In addition, dark-field illumination provides a common means of visualizing autoradiographic grains but tictorial stains are not optimally visible under these conditions. In an effort to find a counterstain that would be compatible with dark-field illumination, we have investigated the use of fluorescent dyes. Of the fluorescent dyes tested, bisbenzimide (Hoechst 33258) in pH 2.0 buffer was found to be optimal. Bisbenzimide counterstaining gave good resolution of cellular boundaries and appeared not to interfere with the ability to visualize autoradiographic grains. Furthermore, the illumination of bisbenzimide and of the autoradiographic grains could be controlled indepe...Continue Reading

References

Jan 1, 1989·Comparative Biochemistry and Physiology. A, Comparative Physiology·P C Emson
Aug 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·M SchallingD H Schlesinger
Mar 1, 1989·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·D F Clayton, A Alvarez-Buylla
Aug 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·G R UhlJ F Habener
Feb 1, 1982·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·L C SchmuedP E Sawchenko

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Citations

Jun 22, 2000·The Journal of Comparative Neurology·R Ubink, T Hökfelt
Dec 10, 1996·Proceedings of the National Academy of Sciences of the United States of America·Z Q XuT Hökfelt
Apr 22, 2004·The Journal of Comparative Neurology·Stephen A Schnell, Martin W Wessendorf
Jan 24, 2006·General and Comparative Endocrinology·Seong W KangMohamed E El Halawani
Jan 3, 2001·The Journal of Comparative Neurology·H Wang, M W Wessendorf
May 8, 1999·The American Journal of Physiology·W C Engeland, B K Levay-Young

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