Grant-in-Aid for Scientific Research (C)
|Allocation Type||Single-year Grants|
General anatomy (including Histology/Embryology)
|Research Institution||TOTTORI UNIVERSITY|
IINO Akihiro TOTTORI UNIV., FACUL.MEDCINE,PROFESSOR, 医学部, 教授 (50031969)
INAGA Sumire TOTTORI UNIV., FACUL.MEDCINE,RESEARCH ASSOCIATE, 医学部, 助手 (60116358)
NAGURO Tomonori TOTTORI UNIV., FACUL.MEDCINE,ASSOCIATE PROFESSOR, 医学部, 助教授 (50032230)
TANAKA Keiichi SEIREI CHRISTOPHER COLLEG.NURS.
FUNAKI Kenji SHIMANE UNIV.
舟木 賢治 鳥取大学, 医学部, 助手 (90091579)
|Project Period (FY)
1995 – 1997
Completed(Fiscal Year 1997)
|Budget Amount *help
¥2,400,000 (Direct Cost : ¥2,400,000)
Fiscal Year 1997 : ¥300,000 (Direct Cost : ¥300,000)
Fiscal Year 1996 : ¥300,000 (Direct Cost : ¥300,000)
Fiscal Year 1995 : ¥1,800,000 (Direct Cost : ¥1,800,000)
|Keywords||CHROMOSOME / HYDROUS MATERIALS / LOW VACUUM SEM / CHROMATIN STRUCTURE / HELICAL COIL / CHROMOSOME BAND / COOL STAGE / PLATINUM BLUE STAINING / 反射電子像 / 高次コイル構造 / 含水標本 / 反射電子 / 核|
Using isolated hydrous chromosomes, we investigated the higher order structure of metaphase chromosome by low vaccum scanning electron microscopy (LVSEM) in this study. In the first, we developed new techniques for observing the hydrous biological materials such as human chorionic villi, erythrocytes in the blood vessel, macrophages and HeLa cells by LVSEM.Especially, mitochondrial cristae in a HeLa cell were found first time by LVSEM.Next, we observed the ultrastructures of nucleus and chromosomes of cultured Chinese hamster bone marrow cells by the new techniques.
Sucsessfully, hierarchical helical coils and band-like structure were observed obviously on hydrous metaphase chromosomes by LVSEM.
1. New techniques for observing hydrous biological materials by LVSEM
Platinum blue (Pt-blue) staining method and the use of cooling stage combined with the pretreatment with DMSO were developed for observing hydrous biological materials by LVSEM (-10ﾟC,270 Pa) in cooperation with Professor Tanaka (SEIREI Christopher College of Nursing).
These new techniques were contributed to study not only about chromosomes but also about general biological hydrous materials.
2. Higher order structure of hydrous chromosomes
Fixed metaphase chromosomes showed interesting figures in the hydrous condition by LVSEM.Hierarchical helical coils were recognized on isolated hydrous chromosomes from Chinese hamster cells (bone marrow cells and lung fibroblast cells) at a magnification of 20,000.
In addition to this, band-like structures were observed unexpectedly on hydrous metaphse chromosomes as a contrast of backscattered electron (BSE) signals depended on Pt-blue staining.
These results suggested the native higher order structure of metaphase chromosome, and we found a clue to the basic investigation of the banding formation mechanism of chromosome by electron microscopy.