Se. Mahoney et al., 3-DIMENSIONAL LASER-SCANNING CONFOCAL MICROSCOPY OF IN-SITU HYBRIDIZATION IN THE SKIN, The American journal of dermatopathology, 16(1), 1994, pp. 44-51
In situ hybridization is an important tool in molecular and developmen
tal biology to detect specific nucleic acid sequences (either mRNA or
DNA) within cells. This technique is especially applicable to tissue s
ections since it provides information about the spatial distribution o
f DNA or mRNA sequences. However, previous studies utilizing in situ h
ybridization in the skin were hampered by a high degree of nonspecific
background, which has made interpretation of the results difficult. I
n this paper, we demonstrate how refinements in in situ hybridization
techniques, combined with laser-scanning confocal microscopy, signific
antly reduce nonspecific background and produce improved resolution of
in situ hybridization in skin specimens. The sensitive detection meth
od of laser-scanning microscopy allows three-dimensional localization
of S-35 radioactive-labeled riboprobes within the emulsion of specimen
s, which is not possible with conventional bright or dark field light
microscopy.