Abstract: Systems and methods of microscopy include and/or implement a laser configured to output a first beam and a second beam; a first optical system configured to receive the first beam, the first optical system comprising: a first beam splitter configured to split the first beam into a first component and a second component, a first photonic crystal fiber configured to modify a bandwidth of the first component, and a first pulse shaper configured to shape the first component in at least one of a spatial aspect or a temporal aspect, the first pulse shaper including a first diffraction grating, a first achromatic half-wave plate, a first lens, and a first two-dimensional spatial light modulator (SLM); and a second optical system configured to receive the second beam.

Abstract: A photon peak event detection system accepts an analog output from a photon sensor, directly digitizes the analogy output and includes a graphics processing unit (GPU) programmed to conduct a photon peak event detection in real-time via a photon count program that analyzes the digitized photon sensor output in sampling periods each having at least three consecutive data points to determine a local maximum among the consecutive data points and compare the local maximum to one or more predetermined thresholds to determine whether or not a photon was received in each sampling period, the algorithm providing photon counts to a phasor analysis program in the GPU. The phasor analysis program calculates pixelwise fluorescence lifetime phasor data in real-time and sends the data to a central processing unit.