A novel technique for quantitative immunohistochemical imaging of various neurochemicals in a multiple-stained brain slice.
Here we describe a novel technique for comparative analysis of the distributions of various neurochemicals visualized using multiple immunohistochemistry in the same brain slice. As an example, the distributions of tyrosine hydroxylase, substance P and glutamate decarboxylase in coronal slices of rat brains were compared. Each slice was divided into approximately 220,000-300,000 microareas at 20-microm intervals, and the immunohistochemical intensities of the three substances in each microarea were analyzed independently using a brain mapping analyzer; a microphotometry system previously developed in our laboratory (Sutoo et al., J. Neurosci. Methods, 1998; 85: 161-73). No significant differences between the distribution of each substance were observed in single- and triple-labeled slices. We believe that this method will facilitate the investigation of the functions of the central nervous system and the disorders thereof in various diseases.
Institute of Medical Science, University of Tsukuba, Tsukuba 305-8575, Japan. dsutoo@md.tsukuba.ac.jp
Den'etsu
D
Sutoo
Kayo
K
Akiyama
Kazunori
K
Yabe
D000818:Animals; D001921:Brain / Q000378:metabolism*; D001923:Brain Chemistry; D005455:Fluorescent Antibody Technique / Q000379:methods; D005968:Glutamate Decarboxylase / Q000378:metabolism; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D013373:Substance P / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D013373:Substance P; D014446:Tyrosine 3-Monooxygenase; D005968:Glutamate Decarboxylase