Bioengineering Seminar Series: Michael J. Levene

Friday, March 1, 2013
11:00 a.m.-12:00 p.m.
Room 1200, Jeong H. Kim Engineering Building
Professor Ian White
ianwhite@umd.edu

Getting Deeper with Multiphoton Microscopy: In Vivo Neuroscience and 3D Histopathology

Michael J. Levene
Associate Professor
Biomedical Engineering and Neurology
Yale University and Yale School of Medicine

Multiphoton microscopy has found wide application due in part to its ability to image deeper into scattering tissues than alternative approaches. However, it is still in general limited to a few hundred microns penetration depth. My lab has been developing micro-optics and optical clearing methods to extend the range of multiphoton microscopy and open up new application areas for in vivo neuroscience and 3D histopathology.

Optical techniques are revolutionizing neuroscience, from multiphoton imaging of Ca2+- and voltage-sensitive dyes to direct stimulation of neurons with optogenetics, yet, gaining high-resolution optical access to deeper layers of cortex and sub-cortical regions remains a challenge. We have been developing invasive micro-optics, including both gradient index (GRIN) lenses and micro-prisms, for in vivo multiphoton microscopy of deep structures in mouse. I will show recent results from our lab, including using micro-prisms, which give simultaneous access to all 6 cortical layers in vivo, to image neural activity in V1 of awake mice.

Histological evaluation of tissue samples is the backbone of disease diagnosis, yet pathologists have remained mired in 2 dimensions. My lab has been working to bring pathology into the 3rd dimension through complete imaging of entire resected tissue samples using multiphoton microscopy and optical clearing. I will present multiphoton images of human biopsies that are 1.5 mm thick. Combining intrinsic fluorescence, second harmonic generation, and the use of extrinsic dyes provides contrast similar to standard histological stains. Further, I will show that optical clearing and multiphoton microscopy are compatible with traditional approaches to histology, enabling the use of specialty stains and immunohistochemistry where necessary.

Audience: Graduate  Faculty  Post-Docs 

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