Journal
JOURNAL OF BIOMEDICAL OPTICS
Volume 13, Issue 4, Pages -Publisher
SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS
DOI: 10.1117/1.2967983
Keywords
flow cytometry; ultrafast; multiphoton; extended cavity; green fluorescent protein (GFP); noninvasive
Funding
- NCI NIH HHS [P50 CA093990, P50 CA93990] Funding Source: Medline
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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. (C) 2008 Society of Photo-Optical Instrumentation Engineers.
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