Departments of Chemical Engineering and Anatomy, University of Queensland, St. Lucia, Queensland 4067, Australia.
Appl Environ Microbiol. 1983 Sep;46(3):716-21. doi: 10.1128/aem.46.3.716-721.1983.
A technique is described for the quantitative determination of the distributed growth of Saccharomyces cerevisiae immobilized in polyacrylamide gel. Gel specimens were embedded in paraffin or gelatin and paraffin before sectioning and staining. Photomicrographs of specimen sections were enlarged, and cell microcolony volumes were determined as a function of position in the gel by grid transparency analysis. Overall cell densities within the gel were calculated for a quantitative comparison with values measured by a second spectrophotometric method. The results show good agreement and demonstrate the sigmoidal growth of the immobilized cells, reaching a maximum steady-state value. The technique shows promise as a general method for following the transient growth of organisms immobilized within gel particles.
介绍了一种定量测定固定在聚丙烯酰胺凝胶中的酿酒酵母分布生长的技术。凝胶标本在切片和染色前用石蜡或明胶和石蜡包埋。对标本切片的显微照片进行放大,并通过网格透明度分析确定细胞微菌落体积作为凝胶中位置的函数。计算凝胶内的总细胞密度,以便与通过第二种分光光度法测量的值进行定量比较。结果表明,该技术具有良好的一致性,并证明了固定细胞的S 形生长,达到了最大稳态值。该技术有望成为一种通用方法,用于跟踪固定在凝胶颗粒内的生物体的瞬态生长。