In-line spectroscopic cytometry for real-time monitoring of celllular and/or molecular bioprocesses

Abstract

An analytical method and system for monitoring cellular status wherein excitation energy is focused into an excitation cytometry volume within a cell sample that is located within a bioreactor or fermenter to induce optical signals from intracellular compounds. The resulting optical signals are directed to a detection subsystem that has at least two detection channels. One channel detects the elastic (Rayleigh or Mie) scattering signal from the cell that identifies the presence of the cell within the excitation volume. Another channel detects the fluorescence and/or Raman scattering signal of intracellular compounds of the cells. The elastic scattering signals associated with individual cells are used as event-triggers to gate the detection of the intracellular fluorescence and/or Raman signal to eliminate the background noises from the suspension media. The intracellular fluorescence and/or Raman scattering signal are used to characterize cellular status based on the fluorescence intensities at specific fluorescence wavelengths and peak analysis of Raman scattering spectra.

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