Spectral measurements of large particles by flow cytometry

Abstract

Flow cytometers designed to analyze large particles are enabling new applications in biology and chemistry. Similarly, flow spectroscopy approaches are extending the capabilities of the flow cytometry platform. Here, we report on the adaptation of a commercial large particle analyzer to measure fluorescence and Raman spectra of individual particles at high speeds. We modified a Union Biometrica COPAS Plus instrument to allow red excitation and optical fiber-based light collection and spectral analysis using a spectrograph and CCD array detector. These modifications did not compromise the ability of the instrument to resolve different sized particles based on their extinction and time of flight signals. The modified instrument has the sensitivity and spectral resolution to measure the fluorescence and Raman signals from individual particles with signal integration times of 10 usec. The high speed spectral analysis of individual particles in flow will enable new applications in biological and chemical analyses.