Quantitative Image Analysis of the Central Monoamine Fluorescent Tissue

• Published in: Japanese Journal of Pharmacology 1990, Vol.52 (suppl-1.1), p.196-196
• Main Authors: Sato, Katsuhiko, Hayashi, Masaaki, Hara, Ichie, Masuda, Mikio, Takada, Kaori, Kawamura, Kenji, Takahashi, Noriyuki, Shibuya, Takeshi
• Format: Article
• Language: English
• ISSN: 0021-5198; 1347-3506
• DOI: 10.1016/S0021-5198(19)55410-5

Out of the morphological studies regarding the nervous system in the field of neuroscience is a method for making the transmitter directly visible. This method has already been put to practical use for catecholamine(CA) and serotonin(5-HT). However, quantitation of the fluorescent substance in a tissue slice has posed a problem in histochemical studies. We have assessed the fluorescent intensities of fluorescent tissue specimens of central CA and 5-HT by determining color intensity with an image analyzing device (Imaging Research Inc., Canada). Male Wistar strain rats were used in this experiment, and fluroresent tissue specimens were prepared from cerebral tissue using the formaldehyde method. To establish a fluorescent standard, a fluorescent specimen was prepared by adding CA and 5-HT in concentrations of 0.5, 5 and 10pg/μl to a 1% albumin solution. Thereafter, we could establish conversion tables for their optical densities and fluorescent intensities. To do an image analysis, we prepared lighter and shaded images of the specimen with a CCD camera by placing a fluorescent tissue film on an illuminator. We analyzed the fluorescent intensity quantitatively by making false color indications(256 colors) according to light and shade levels. These results prove that a fluorescent histochemical judgement of the central CA and 5-HT which had previously proved to be difficult could be done quantitatively by applying this method.