Development of the in vivo flow cytometer

An in vivo flow cytometer has been developed that allows the real-time detection and quantification of circulating cells containing fluorescent proteins or labeled with fluorochrome molecules in live animals, without the need to extract blood samples. A stationary laser beam is focused by a cylindrical lens to a slit of light that is then demagnified and focused across a blood vessel by an achromat and microscope objective. Fluorescent cells are excited one by one as they flow through the excitation laser light slit, creating a burst of fluorescence whose width is inversely proportional to their velocity. The fluorescence signal is detected through a confocal slit aperture using a photomultiplier tube. The analog signal from the photomultiplier tube is then digitized, filtered, and recorded as a function of time onto a computer. Computer programs post- process the data for the presence of cell signal, as well as various aspects of the cell signal such as height, width, and temporal location of the signal peak. Two in vivo flow cytometers have been built: a single-slit, single-color system and a two- slit, two-color system. The single-slit, single-color system provides excitation at 632 nm, and the two-slit, two-color system provides excitation at 632 nm and 473 nm. The two- slit, two-color system can operate in several different modes: single-slit at 632 nm or 473 nm, double-slit at 632 nm or 473 nm, and double-slit with one excitation slit at 632 nm and the other at 473 nm.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Mechanical Engineering.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2004

Author dc:creator

• Novak, John P. (John Peter), 1957-

Advisor dc:contributor.advisor

• Charles Lin.

• 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/1721.1/30331

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/30331