Single-cell viability assessment with a novel spectro-imaging system.
Single-cell viability assessment by means of plural dye probes require the spectral and temporal analysis of microscopic images of the test cells. To meet this requirement, we have developed a simple and compact spectro-imaging system using an image slicer and a grism. The image slicer was made of a bundle of 100 optical fibers. The field of view is divided into 10 x 10 sections. The spectral data of each section could be recorded every 5 s in the range from 400 to 800 nm at 5 nm resolution. The viability changes of yeast or tobacco single-cells were measured with this system. Using BY-2 cells, for example, the response to a chemical stress of saponin was measured by means of two fluorescent probes. The spectral-spatial-temporal data of fluorescein and DNA bound ethidium bromide provided us with useful information about the dynamic change of cell membrane permeability from which the cell viability was assessed.
Matsuoka
Hideaki
H
Department of Biotechnology and Life Science, Faculty of Technology, Tokyo University of Agriculture and Technology, 2-24-16, Nakamachi, Koganei, Tokyo 184-8588, Japan. bio-func@cc.tuat.ac.jp
Kosai
Yuri
Y
Saito
Mikako
M
Takeyama
Norihide
N
Suto
Hiroshi
H
D000666:Amphotericin B / Q000633:toxicity; D000935:Antifungal Agents / Q000633:toxicity; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003592:Cytophotometry / Q000295:instrumentation* / Q000379:methods*; D004996:Ethidium / Q000737:chemistry; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000166:cytology; D012503:Saponins / Q000633:toxicity; D013050:Spectrometry, Fluorescence / Q000295:instrumentation* / Q000379:methods*; D013997:Time Factors; D014026:Nicotiana / Q000166:cytology / Q000187:drug effects
D000935:Antifungal Agents; D005452:Fluoresceins; D005456:Fluorescent Dyes; D012503:Saponins; D000666:Amphotericin B; D004996:Ethidium