University of California

Mihaela Balu

Bruce J Tromberg

Hideharu Mikami

Eric Olaf Potma

The disclosure provides noninvasive methods for the in vivo identification and characterization of various immune cell populations in human skin. The methods of the disclosure employ advanced imaging systems based on nonlinear optical microscopy to generate images that are analyzed to detect, identify, differentiate, and quantify immune cell populations in human skin by their morphological, metabolic and behavioral signatures.

Methods for non-invasive, label-free imaging of cellular immune response in human skin using a nonlinear optical microscopy imaging system

A multiphoton microscope based on two-photon excited fluorescence and second-harmonic generation that images FOVs of about 0.8 mm(without stitching adjacent FOVs) at speeds of 10 frames/second (800&#xd7;800 pixels) with lateral and axial resolutions of 0.5 &#x3bc;m and 2.5 &#x3bc;m, respectively. The scan head of the instrument includes a fast galvanometric scanner, relay optics, a beam expander and a high NA objective lens. The system is based on a 25&#xd7;, 1.05 NA water immersion lens, which features a long working distance of 1 mm. A proper tailoring of the beam expander, which consists of the scan and tube lens elements, enables scaling of the FOV. The system and method also include a flat wavefront of the beam, minimum field curvature, and suppressed spherical aberrations. All aberrations in focus are below the Marechal criterion of 0.07&#x3bb; rms for diffraction-limited performance.

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Irvine, CA, United States of America

Mihaela Balu