Systems and methods for classifying and/or sorting T cells by activation state are disclosed. The system includes a cell classifying pathway, a single-cell autofluorescence image sensor, a processor, and a non-transitory computer-readable memory. The memory is accessible to the processor and has stored thereon a trained convolutional neural network and instructions. The instructions, when executed by the processor, cause the processor to: a) receive the autofluorescence intensity image; b) optionally pre-process the autofluorescence intensity image to produce an adjusted autofluorescence intensity image; c) input the autofluorescence intensity image or the adjusted autofluorescence intensity image into the trained convolutional neural network to produce an activation prediction for the T cell.