SUBCELLULAR-LOCALIZATION OF ALPHA-TUBULIN AND OPSIN MESSENGER-RNA IN THE GOLDFISH RETINA USING DIGOXIGENIN-LABELED CRNA PROBES DETECTED BY ALKALINE-PHOSPHATASE AND HRP HISTOCHEMISTRY
Lk. Barthel et Pa. Raymond, SUBCELLULAR-LOCALIZATION OF ALPHA-TUBULIN AND OPSIN MESSENGER-RNA IN THE GOLDFISH RETINA USING DIGOXIGENIN-LABELED CRNA PROBES DETECTED BY ALKALINE-PHOSPHATASE AND HRP HISTOCHEMISTRY, Journal of neuroscience methods, 50(2), 1993, pp. 145-152
This paper describes a method for non-radioactive in situ hybridizatio
n providing subcellular localization of mRNA in 3 mu m cryosections. W
e used two alternative colorimetric reactions to detect digoxigenin-la
beled cRNA probes: alkaline phosphatase and HRP (horseradish peroxidas
e). With some probes the signal with the alkaline phosphatase reaction
was intense, and diffusion of the reaction product was noticeable. Us
ing HRP-conjugated antibodies improved the resolution but decreased th
e sensitivity of the signal. Photoamplification of the HRP reaction pr
oduct increased the contrast and improved the sensitivity of the techn
ique.