al-Rubeai M A, Musgrave S C, Lambe C A, Walker A G, Evans N H, Spier R E
Department of Microbiology, University of Surrey, Guildford, UK.
Enzyme Microb Technol. 1990 Jun;12(6):459-63. doi: 10.1016/0141-0229(90)90058-x.
Rapid and reliable methods for the determination of survival, proliferation, and metabolic activity of immobilized cells in gels are described. The first method is based on an MTT assay that measures qualitatively and quantitatively the metabolic activity of the cells. The second method determines cell number by measuring the amount of DNA available for Feulgen staining. In the third method, two fluorescent dyes are used to differentially stain viable and dead cells. The fourth method involves the use of glutaraldehyde to protect the cells when melting the gel to facilitate hemocytometric count. The presented techniques should help to test the efficiency of the immobilization procedures and to monitor the growth and survival of immobilized cells.
本文描述了用于测定凝胶中固定化细胞的存活、增殖和代谢活性的快速且可靠的方法。第一种方法基于MTT 法,可定性和定量地测量细胞的代谢活性。第二种方法通过测量可用于福尔根染色的DNA量来确定细胞数量。在第三种方法中,使用两种荧光染料对活细胞和死细胞进行差异染色。第四种方法是在融化凝胶以利于血细胞计数时,使用戊二醛保护细胞。所提出的技术应有助于测试固定化程序的效率,并监测固定化细胞的生长和存活情况。