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.
Sutoo
Den'etsu
D
Institute of Medical Science, University of Tsukuba, Tsukuba 305-8575, Japan. dsutoo@md.tsukuba.ac.jp
Akiyama
Kayo
K
Yabe
Kazunori
K
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