Automated instrument for measuring biological transport kinetics over intervals of a few seconds
✍ Scribed by Olen R. Brown; Jerry Barton
- Publisher
- John Wiley and Sons
- Year
- 1974
- Tongue
- English
- Weight
- 558 KB
- Volume
- 16
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
Abstract
Automated sampling device was designed to permit determination of rates of biological transport of metabolites into cells. The substrate was automatically introduced into a stirred cell suspension at 37°C. The first sample was automatically taken after a mixing interval of 1 sec and nine subsequent samples were taken at programmable intervals (1 to 100 sec). The samples were forced by pressure differential (vacuum) through 0.4 μ pore size membranes and approximately 50 μl were collected in disposable cups. The duration of the sampling interval was controllable down to 0.1 sec. The samples preserved records of the substrate concentrations in solution at the time of filtration. With the use of suitable radioactive labeled isotopes, the changes in substrate concentrations may be conveniently measured by liquid scintillation spectrometry, but other analytical procedures of suitable sensitivity may be used. Initial and steady‐state transport rates of succinate and glucose in Escherichia coli were obtained using the device.