1996 Fiscal Year Final Research Report Summary
Application of high pressure freezing techinque
| Project/Area Number |
07670026
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General anatomy (including Histology/Embryology)
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| Research Institution | Kagoshima University |
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Principal Investigator |
MURATA Fusayosi Fuculty of Medicine Kagoshima University Professor, 医学部, 教授 (60020765)
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| Co-Investigator(Kenkyū-buntansha) |
ORMORI Jun Fuculty of Medicine Kagoshima University Reseach Associate, 医学部, 助手 (10281221)
TSUYAMA Shinichiro Fuculty of Medicine Kagoshima University Associate professor, 医学部, 助教授 (30041346)
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| Project Period (FY) |
1995 – 1996
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| Keywords | high pressure freezing / freeze substitution / histochemistry / freeze fracture / immunohistochemistry / lectin / glycoconjugate / digestive tract |
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| Research Abstract |
High pressure freezing technique is a new technique in histology, histochemistry, immunohistochemistry and cell biology. Compared to well used rapid freezing technique, this new method can get wider ideal vitrification area. The depth of this ideal area is more than ten times wider than that obtained by rapid freezing technique. This makes it possible to observe whole large cells such as nerve cells or oocytes, whick rapid freezing technique could not achieve.
There are two main approaches to apply this method. One is to observe histochemical reactions in combination with freeze substituion method Acrolein and acetone were used in our experiments, It became apparent that the former was a excellent method to observe histochemical reaction or immunohistochemical reaction. Good results have been obtained for the study of glycoconjugate histochemistry and H-K-ATPase immunocytochemitry in digestive tract cells.
Another method is to observe ultrastructure of cells in combination with freeze fracture technique. This is the joint work of our group and department of anatomy, Yamanashi medical college (Prof. S.Ohno). In this experiment, it became apparent that good cell preservation was kept even without fixation in many cell types of digestive tract.
In addition to such investigation, we undertook two enlightment works for high pressure freezing. One was the wet laboratory experiment in collabaration with Kyushu district of Japanese society of electron microscopists and the second was the speech at summer school of Japanese society of electron microscopists.
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