We present a wide-field fluorescence lifetime imaging (FLIM) system with optical sectioning by structured illumination microscopy (SIM). FLIM measurements were made using a time gated ICCD camera in conjunction with a pulsed nitrogen dye laser operating at 450 nm. Intensity images were acquired at multiple time delays from a trigger initiated by a laser pulse to create a wide-field FLIM image, which was then combined with three phase SIM to provide optical sectioning. Such a mechanism has the potential to increase the reliability and accuracy of the FLIM measurements by rejecting background intensity. SIM also provides the opportunity to create volumetric FLIM images with the incorporation of scanning mechanisms for the sample plane. We present multiple embodiments of such a system: one as a free space endoscope and the other as a fiber microendoscope enabled by the introduction of a fiber bundle. Finally, we demonstrate the efficacy of such an imaging system by imaging dyes embedded in a tissue phantom.
Taylor Hinsdale, Bilal H. Malik, Jose J. Rico-Jimenez, Javier A. Jo, and Kristen C. Maitland, "Optically sectioned wide-field fluorescence lifetime imaging endoscopy enabled by structured illumination
(Conference Presentation)," Proc. SPIE 9713, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIII, 97130X (Presented at SPIE BiOS: February 16, 2016; Published: 27 April 2016); https://doi.org/10.1117/12.2213599.4848767273001.
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