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一种用于测量细胞悬液耗氧量的新型光度法。

A new photometric method for oxygen consumption measurements in cell suspensions.

作者信息

Mueller-Klieser W, Zander R, Vaupel P

出版信息

J Appl Physiol (1985). 1986 Aug;61(2):449-55. doi: 10.1152/jappl.1986.61.2.449.

Abstract

A new technique is described for measuring O2 consumption rates and O2 concentrations in suspensions of respiring cells. Aliquots of a cell suspension kept in a special thermostated precision syringe are injected into the measuring system in defined time intervals. The O2 content of these samples is determined photometrically, as reported previously. The O2 consumption per cellular wet weight and/or per single cell can be calculated from the cell volume fraction, the physical density, the cell concentration in the suspension, and the time-dependent decline of the O2 concentration in the precision syringe. The minimum detectable amount of O2 is 0.1 microliter O2, which corresponds to 0.001 (vol/vol) of O2 if a 100-microliters sample of suspended cells is analyzed. Reproducibility of the O2 consumption measurement is 9% of the measured value. The advantages offered by this method are the straightforward calibration in absolute terms, the short time required for one analysis (2-6 min), a high sensitivity, the simultaneous determination of overall O2 concentration and O2 consumption rates in cell suspensions, and the great variability in the application.

摘要

本文描述了一种用于测量呼吸细胞悬液中氧气消耗率和氧气浓度的新技术。将保存在特殊恒温精密注射器中的细胞悬液等分试样,按规定的时间间隔注入测量系统。如先前报道的那样,通过光度法测定这些样品的氧气含量。根据细胞体积分数、物理密度、悬液中的细胞浓度以及精密注射器中氧气浓度随时间的下降情况,可以计算出每细胞湿重和/或每单个细胞的氧气消耗量。氧气的最低可检测量为0.1微升氧气,如果分析100微升的悬浮细胞样品,这相当于0.001(体积/体积)的氧气。氧气消耗测量的重现性为测量值的9%。该方法的优点包括绝对校准直接明了、一次分析所需时间短(2 - 6分钟)、灵敏度高、可同时测定细胞悬液中的总氧气浓度和氧气消耗率以及应用的多样性。

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