Related Experiment Videos
Rapid scanning thermal lens/laser transmission densitometer
1Department of Chemistry, University of Michigan, Ann Arbor 48109.
Applied and Theoretical Electrophoresis : the Official Journal of the International Electrophoresis Society
|January 1, 1988
Summary
An automated densitometer uses thermal lens and laser transmission principles. Thermal lens densitometry offers lower detection limits for analyzing proteins, while transmission densitometry is better for high optical densities.
Area of Science:
- Analytical Chemistry
- Biophysical Chemistry
Background:
- Densitometry is crucial for quantifying stained biomolecules after separation.
- Existing methods like laser transmission densitometry have limitations at very low or high concentrations.
Purpose of the Study:
- To describe an automated densitometer combining thermal lens and laser transmission techniques.
- To compare the performance of thermal lens densitometry versus transmission densitometry.
Main Methods:
- Development of a novel automated densitometer.
- Utilizing the thermal lens effect for sensitive detection.
- Employing conventional laser transmission for broader dynamic range.
- Characterization using proteins separated by SDS-PAGE and stained with Coomassie Brilliant Blue G250.
Main Results:
- The thermal lens densitometer achieved lower detection limits compared to transmission mode.
- Transmission densitometry proved more suitable for samples with high optical densities.
- The instrument demonstrated effective characterization of stained proteins.
Conclusions:
- The dual-mode densitometer offers versatility for protein analysis.
- Thermal lens densitometry is advantageous for trace-level detection.
- Transmission densitometry remains valuable for quantifying concentrated samples.