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Related Experiment Videos

Violet laser diodes as light sources for cytometry.

H M Shapiro1, N G Perlmutter

  • 1Howard M. Shapiro, M.D., P.C., West Newton Massachusetts 02465-2513, USA. hms@shapirolab.com

Cytometry
|May 30, 2001
PubMed
Summary

Violet laser diodes offer a cost-effective and compact solution for DNA fluorescence measurement. These diodes efficiently excite DAPI and Hoechst dyes, enabling precise DNA content analysis in flow cytometry.

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Area of Science:

  • Biophotonics
  • Analytical Chemistry
  • Cell Biology

Background:

  • Violet laser diodes (395-415 nm) are now commercially available, offering compact and efficient light sources.
  • These systems, initially expensive, integrate diodes with optics, power supplies, and cooling for stable output.

Purpose of the Study:

  • To evaluate the utility of violet laser diodes in a flow cytometer for DNA content analysis.
  • To assess the performance of DAPI and Hoechst dyes with violet laser excitation.

Main Methods:

  • A 4-mW, 397-nm violet diode system was integrated into a laboratory-built flow cytometer.
  • Fluorescence of DAPI and Hoechst dyes in cells was measured, along with forward and orthogonal light scattering.

Main Results:

  • High-precision DNA content histograms were achieved using DAPI staining (CV 1.7%).
  • Hoechst 33342 showed lower precision, while Hoechst 34580 provided high fluorescence but degraded.
  • Light scatter signals displayed relatively broad distributions.

Conclusions:

  • Violet laser diodes are suitable, affordable, and efficient light sources for DNA fluorescence measurements.
  • These diodes effectively excite DAPI and Hoechst dyes and have potential for other fluorescent probes.

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