High throughput 3-D tissue cytometry

This thesis presents the ongoing technological development of high throughput 3-D tissue cytometry.and its applications in biomedicine. 3-D tissue cytometry has been developed in our laboratory based on two-photon microscopy (TPM) that is capable of in situ 3-D imaging of tissue up to a volume of several mm3 with subcellular resolution. This high throughput tissue cytometry achieves an imaging rate of 2 mm3/hour. I optimize the performance of this instrument by developing two new techniques. First, the image signal-to-noise-ratio and the microscope penetration depth can be improved by reducing tissue scattering. Optical clearing agents can significantly lower tissue scattering by index matching of different tissue constituents. While the application of tissue clearing agent has been extensively studied in fresh tissues, its application has not been extensively studied in paraffin fixed and frozen tissues. Frozen tissues are particularly important as tissue sections can be retained for biochemical and genetic analysis after fluorescence imaging. We investigate the effects of optical clearing agents in sub-zero temperature in terms of TPM image contrast and penetration depth. Second, tissue cytometry are often used in the detection of rare cells.

Department dc:contributor.department

Massachusetts Institute of Technology. Department of Mechanical Engineering

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2007

Author dc:creator

• Kwon, Hyuk-Sang, 1971-

Advisor dc:contributor.advisor

• Peter T.C. So.

• Mechanical Engineering.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/39903

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/39903