University of Notre Dame Du Lac

Yide Zhang

Scott Howard

Genevieve Vigil

Cody J Smith

One aspect of the present disclosure relates to systems and methods that can be used for super-sensitivity multiphoton frequency-domain fluorescence lifetime imaging microscopy (MPM-FD-FLIM). One or more laser light sources can deliver excitation light with an intensity modulated by a periodic function to a sample, which includes a fluorophore concentration. A receiver can receive a fluorescence signal emitted from the fluorophore concentration over a time period. A computing device can transform the fluorescence signal to the frequency domain and measure a fluorescence lifetime associated with the sample in the frequency domain based on a DC component and at least one higher harmonic component generated by non-linear optical mixing of the modulated excitation light. Using the DC component and the at least one higher harmonic component can increase the sensitivity of an image created by the MPM-FD-FLIM system.

Super-sensitivity multiphoton frequency-domain fluorescence lifetime imaging microscopy

A microscopy method and system includes obtaining a plurality of raw fluorescence images, respective ones of the plurality of raw fluorescence images obtained by exciting a sample using a light source at a respective irradiance value in a weak saturation region of the sample, and capturing the respective raw fluorescence image; applying a plurality of weights to the plurality of raw fluorescence images in a one-to-one correspondence so as to generate a plurality of weighted fluorescence images, wherein a respective weight is based on the respective irradiance value at which the corresponding raw fluorescence image was obtained; and linearly combining the plurality of weighted fluorescence images, thereby generating the output image having a resolution greater than a diffraction limit. Respective raw fluorescence images correspond to irradiance values different from one another.

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South Bend, IN, United States of America

Yide Zhang