1995 Fiscal Year Final Research Report Summary
DETECTION OF ENZYME ACTIVITIES ON ULTRATHIN FROZEN SECTIONS - A NEW METHOD FOR ELECTRON MICROSCOPIC ENZYME HISTOCHEMISTRY.
| Project/Area Number |
05670030
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
General anatomy (including Histology/Embryology)
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| Research Institution | JIKEI UNIVERSITY SCHOOL OF MEDICINE |
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Principal Investigator |
FUKUSHIMA Osamu JIKEI UNIVERSITY SCHOOL OF MEDICINE,LECTURER, 医学部, 講師 (60173332)
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| Project Period (FY) |
1993 – 1995
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| Keywords | ULTRATHIN FROZEN SECTION / ENZYME HISTOCHEMISTRY / ACID PHOSPHATASE / ALKALINE PHOSPHATASE / AMINOPEPTIDASE / DIPEPTIDYL PEPTIDASE / CYTOCHROME C OXIDASE / cytochrome Coxidase |
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| Research Abstract |
The aim of electron microscopic enzyme histochemistry is to localize the sites of enzyme activities on cellular ultrastructure on order to understand the metabolism of cells. The routine enzyme histochemical techniques employed 40 um sections for demonstrating activities. Our study shows that there are penetration problems in 40 um sections for detection of enzyme activities, especially cytochrome C oxidase activity, and concluded that the ultrathin frozen section is suitable for demonstrating it at electron microscopic level. The study indicates that a histochemical reaction should be done on an ultrathin section. There are two methods for histochemical reactions ; one is an ultrathin frozen section, and the other a plastic ultrathin section. A plastic embedding needs a dehydration procedure. However, our study indicates the displacement of enzyme ptoteins during a dehydration procedure. The ultrathin frozen sections had not been exposed to organic solvents prior to demonstration of enzyme activities. The study shows that an ultrathin frozen section is most suitable for demonstrating enzyme sctivities directly on an ultrathin section. The present study concludes that an ultrathin frozen section should be employed for observation of precise localization of an enzyme activity at electron microscopic level.
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