Observation of single-cell fluorescence spectra in laser flow cytometry

M. R. Gauci*, G. Vesey, J. Narai, D. Veal, K. L. Williams, J. A. Piper

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)

Abstract

We report the first demonstration of directly recording fluorescence spectra of single cells in flow cytometry. An intensified, 512-element photodiode array was used in conjunction with a dispersing prism to capture the fluorescence emission spectra of Coulter ImmunoCheck calibration beads and Dictyostelium discoideum spores stained with the indocarbocyanine derivative CY3, fluorescein isothiocyanate, or R-phycoerythrin. The demonstration was made feasible by enhancing the signal-to-noise ratio of the detection process by using a fast gating technique applied to the detector. Results show that the complete fluorescence spectra of individual stained cells contain information that is normally not captured by conventional flow cytometers. By using the spectrographic flow cytometer, all this information is recorded, allowing small features and shifts in the fluorescence spectra of labelled particles to be studied.

Original languageEnglish
Pages (from-to)388-393
Number of pages6
JournalCytometry
Volume25
Issue number4
DOIs
Publication statusPublished - 1 Dec 1996

Keywords

  • environmental analysis
  • flow cytometry
  • intensfied photodiode array
  • simplified optical multichannel analyser
  • spectral analysis

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