| 摘要 |
PROBLEM TO BE SOLVED: To detect a microorganism in high sensitivity by a flow cytometry capable of computing an accurate viable cell number useful in a wide field by loading applying a nonlethal hydrostatic pressure to a sample containing a microorganism and dyeing the microorganism with a fluorescent coloring matter. SOLUTION: A microorganism (e.g. yeast, etc.), is proliferated in a medium at 24 deg.C, a cell of logarithmic growth phase is collected by centrifugation, resuspended in a medium, put in a plastic tube and the tube is sealed. The sealed tube is put in a pressure vessel made of titanium. A nonlethal hydrostatic pressure in a range of 30-50 MPa is applied to the pressure vessel by a hand pump within two minutes and the vessel is decompressed for about 15 seconds. The microorganism is cultured and dyed in a medium containing a fluorescent coloring matter such as carboxylfluorescein diacetate (CFDA), carboxyldichlorofluorescein diacetate, etc., under pressure of normal pressure to 60 MPa and treated by flow cytometry to compute an accurate viable cell number and to detect the microorganism in high sensitivity. |
