Systems and methods for classifying T cell activation state

A T cell classification device comprising:

• a cell analysis pathway comprising:
• (i) an inlet;
• (ii) an observation zone coupled to the inlet downstream of the inlet, the observation zone configured to present T cells for individual time-resolved autofluorescence interrogation;
• and
• (iii) an outlet coupled to the observation zone downstream of the observation zone;
• a time-resolved autofluorescence decay spectrometer configured to acquire a time-resolved autofluorescence decay signal for a T cell positioned in the observation zone, the time-resolved autofluorescence decay spectrometer comprising a pulsed light source, a photon-counting detector, and photon-counting electronics;
• a processor in electronic communication with the time-resolved autofluorescence decay spectrometer;
• and
• a non-transitory computer-readable medium accessible to the processor and having stored thereon instructions that, when executed by the processor, cause the processor to:
• a) receive the time-resolved autofluorescence decay signal;
• b) compute at least a first phasor coordinate at a first frequency and a second phasor coordinate at a second frequency from the time-resolved autofluorescence decay signal, wherein the first and second frequency are different;
• and
• c) compute an activation prediction for the T cell using at least the first phasor coordinate and the second phasor coordinate.