Electrophoresis with continuous scanning densitometry: Separation of cells in a density gradient

✍ Scribed by N. Catsimpoolas; A.L. Griffith; J.M. Williams; A. Chrambach; D. Rodbard

• Publisher: Elsevier Science
• Year: 1975
• Tongue: English
• Weight: 770 KB
• Volume: 69
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(75)90139-6

✦ Synopsis

An instrument and procedure for electrophoresis with continuous optical scanning densitometry, automated data processing, and related methodology are described for the continuous analysis of electrophoresis and unity gravity sedimentation of macromolecules or cells in a static density gradient system. The instrument consists of a dual-beam spectrophotometer, a scanning stage and scanner control unit, an electrophoresis cell cassette, a filling/purging/cooling module, an analog-to-digital converter, and a digital data-logger. The distribution of cells is monitored repetitively during migration by absorbance measurements at any wavelength in the 200-800-nm range. A computer program provides the statistical analysis of each peak (baseline correction, smoothing, area, mean, standard deviation, skewness, and kurtosis) which can be further utilized for computing additional parameters, such as resolution and heterogeneity. A mixture of human and rabbit erythrocytes were used as a model system to evaluate the performance of the instrument and demonstrate some of its capabilities.