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Two photon fluorescent lifetime imaging is a modality that enables depth-sectioned, molecularly-specific imaging of cells and tissue using intrinsic contrast. However, clinical applications have not been well explored due to low imaging speed and limited field of view, which make evaluating large pathology samples extremely challenging. To address these limitations, we have developed direct temporal sampling two photon fluorescent lifetime imaging (DTS-FLIM), a method which enables a several order of magnitude increase in imaging speed by capturing an entire lifetime decay in a single fluorescent excitation. We use this greatly increased speed to perform a preliminary study using gigapixel-scale imaging of human breast pathology surgical specimens.

We demonstrate a compact, ultrahigh speed spectral-domain optical coherence microscopy (SD-OCM) system for multiscale imaging of specimens at 840 nm. Using a high speed 512-pixel line scan camera, an imaging speed of 210,000 A-scans per second was demonstrated. Interchangeable water immersion objectives with magnifications of 10×, 20×, and 40× provided co-registered en face cellular-resolution imaging over several size scales. Volumetric OCM data sets and en face OCM images were demonstrated on both normal and pathological human colon and kidney specimens ex vivo with an axial resolution of ~4.2 µm, and transverse resolutions of ~2.9 µm (10×), ~1.7 µm (20×), and ~1.1 µm (40×) in tissue. In addition, en face OCM images acquired with high numerical aperture over an extended field-of-view (FOV) were demonstrated using image mosaicking. Comparison between en face OCM images among different transverse and axial resolutions was demonstrated, which promises to help the design and evaluation of imaging performance of Fourier domain OCM systems at different resolution regimes.