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Special Section on Selected Topics in Biophotonics: Diffuse Optics and Optical Molecular Imaging

Extended cavity laser enhanced two-photon flow cytometry

[+] Author Affiliations
Eric R. Tkaczyk, Alan H. Tkaczyk

University of Michigan, Center for Ultrafast Optical Science, Electrical Engineering & Computer Science Department and Michigan Nanotechnology Institute for Medicine and Biological Sciences, Ann Arbor, Michigan 48109

Steve Katnik, Jing Yong Ye

University of Michigan, Center for Ultrafast Optical Science, Electrical Engineering & Computer Science Department, Ann Arbor, Michigan 48109

Kathryn E. Luker, Gary D. Luker

University of Michigan Medical School, Departments of Radiology and Microbiology & Immunology, Ann Arbor, Michigan 48109-0648

Andrzej Myc, James R. Baker

University of Michigan, Michigan Nanotechnology Institute for Medicine and Biological Sciences, Ann Arbor, Michigan 48109-0648

Theodore B. Norris

University of Michigan, Center for Ultrafast Optical Science, Electrical Engineering & Computer Science Department and Michigan Nanotechnology Institute for Medicine and Biological Sciences, Ann Arbor, Michigan 48109

J. Biomed. Opt. 13(4), 041319 (August 28, 2008). doi:10.1117/1.2967983
History: Received September 14, 2007; Revised March 03, 2008; Accepted April 10, 2008; Published August 28, 2008
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We demonstrate enhanced sensitivity in two-photon flow cytometry with an extended cavity laser excitation source. At low power, the home-built 20-MHz oscillator was able to detect a significantly larger fraction, in either phosphate buffered saline (PBS) or whole blood, of green fluorescent protein (GFP)–expressing MCA-207 cells cross-labeled with the membrane-binding lipophilic dye DiD. A geometrical model is used to explain unique features of the signals resulting from the different spatial distribution of DiD and GFP. These unique features include sub-square law scaling of unsaturated two-photon signal, a sigmoidal sensitivity curve for detection under varying powers for cell detection thresholds as low as a single photon, and uncorrelated signal strengths in two detection channels.

Figures in this Article
© 2008 Society of Photo-Optical Instrumentation Engineers

Citation

Eric R. Tkaczyk ; Steve Katnik ; Kathryn E. Luker ; Andrzej Myc ; Theodore B. Norris, et al.
"Extended cavity laser enhanced two-photon flow cytometry", J. Biomed. Opt. 13(4), 041319 (August 28, 2008). ; http://dx.doi.org/10.1117/1.2967983


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