| Project/Area Number |
11670003
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
General anatomy (including Histology/Embryology)
|
|---|
| Research Institution | NIIGATA UNIVERSITY |
|---|
Principal Investigator |
USHIKI Tatsuo School of Medicine, NIIGATA UNIVERSITY, Professor, 医学部, 教授 (40184999)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
HOSHI Osamu School of Medicine, NIIGATA UNIVERSITY, Assistant, 医学部, 助手 (10303124)
|
|---|
| Project Period (FY) |
1999 – 2000
|
| Project Status |
Completed (Fiscal Year 2000)
|
| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2000: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1999: ¥2,200,000 (Direct Cost: ¥2,200,000)
|
|---|
| Keywords | Atomic force microscopy / living cells / cell movement / cytoskeleton / actin |
|---|
| Research Abstract |
The atomic force microscope (AFM) has no lens but simply scans a sharp probing tip over a sample surface, providing topographic images of samples at high resolutions from the micrometer scale to the atomic scale. In this study, we applied AFM to the three-dimensional observation of cultured animal cells in a living state.
We used flat cells attached firmly to the substrate, becauce spherical cells were easily detached from the substrate surface during scanning. To minimize force-induced damages to the cell, imaging force was adjusted to the weakest value. Thus, contact mode AFM of the living cells provided precise information on the shape of cellular processes (spike-like processes, lamellipodia, etc.) at the cellular margin. The contour of cytoskeletal elements just beneath the cell membrane was also clearly observable on the upper surface of the cells.
We then succeeded in obtaining continuously AFM images of living cells for over one hour at time intervals of 2-4 min by using a fluid chamber system. A series of these AFM images were useful for examining the movements of cellular processes in relation to subcellular cytoskeletal elements. Time-lapse movies produced by sequential AFM images also verified the reality of the cellular dynamics.
Wefinally compared the AFM images with images taken by fluorescent microscopy, or scanning electron microscopy. The data indicated the presence of actin accumulation in granular elevations on the leading processes.
|
