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    • 5. 发明申请
    • METHOD AND DEVICE FOR VIABLE AND NONVIABLE PROKARYOTIC AND EUKARYOTIC CELL QUANTITATION
    • 用于可变和不可变原核细胞和原核细胞定量的方法和装置
    • WO0208454A3
    • 2002-06-27
    • PCT/US0123358
    • 2001-07-24
    • GENPRIME INCFLEMING JAMES ESOMES JASON BUCKMCLEAN DARBYHOLCOMB JERAD R
    • FLEMING JAMES ESOMES JASON BUCKMCLEAN DARBYHOLCOMB JERAD R
    • C12Q1/06
    • C12Q1/06Y10T436/101666
    • A rapid method for the quantitation of various live cell types is described. The method may include a variety of steps including: 1) suspending the cells in a detergent-like compound; 2) isolating the washed cells by centrifugation or filtration; 3) resuspending the cells in a solution that contains a preservative, a fluorescent dye and a compound such as dequalinium which can be taken up by the cells; 4) measuring the fluorescence increase over time of the cell-dye mixture with a simple fluorometer; and 5) measuring the native fluorescence of the cells. This new cell fluorescence method correlates with other methods of enumerating cells such as the standard plate count, the methylene blue method and the slide viability technique. The method is particularly useful in several applications such as: a) quantitating bacteria in milk, yogurt, cheese, meat and other foods; b) quantitating yeast cells in brewing, fermentation and bread making; c) quantitating mammalian cells in research, food and clinical settings. The method is especially useful when both total and viable cell counts are required such as in the brewing industry. The method can also be employed to determine the metabolic activity of cells in a sample. The device, and/or system used for cell quantitation is also disclosed.
    • 描述了用于定量各种活细胞类型的快速方法。 该方法可以包括多种步骤,包括:1)将细胞悬浮在洗涤剂状化合物中; 2)通过离心或过滤分离洗涤的细胞; 3)将细胞重悬于包含防腐剂,荧光染料和可被细胞吸收的脱金属化合物的溶液中; 4)用简单的荧光计测量细胞染料混合物随时间的荧光增加; 和5)测量细胞的天然荧光。 这种新的细胞荧光方法与其他枚举细胞的方法相关,例如标准平板计数,亚甲基蓝法和载玻片活力技术。 该方法在几个应用中特别有用,例如:a)定量分析牛奶,酸奶,奶酪,肉类和其他食物中的细菌; b)定量酿造,发酵和面包制作中的酵母细胞; c)在研究,食物和临床环境中定量哺乳动物细胞。 当需要总细胞计数和活细胞计数时,如在酿造工业中,该方法特别有用。 该方法也可用于确定样品中细胞的代谢活性。 还公开了用于细胞定量的装置和/或系统。