Sensing cellular function and molecular activity in vivo using fluorescence lifetime imaging microscopy (FLIM)

Wei Zhong; Dhruv Sud; Mei Wu; Karl A. Merrick; Sofia D. Merajver; David G. Beer; Mary-Ann Mycek

doi:10.1117/12.632912

1 September 2005 Sensing cellular function and molecular activity in vivo using fluorescence lifetime imaging microscopy (FLIM)

Wei Zhong, Dhruv Sud, Mei Wu, Karl A. Merrick, Sofia D. Merajver, David G. Beer, Mary-Ann Mycek

Author Affiliations +

Proceedings Volume 5864, Novel Optical Instrumentation for Biomedical Applications II; 586407 (2005) https://doi.org/10.1117/12.632912
Event: European Conferences on Biomedical Optics 2005, 2005, Munich, Germany

Abstract

In contrast to intensity-based fluorescence microscopy, fluorescence lifetime imaging microscopy (FLIM) bases image contrast on fluorophore excited-state lifetime. This technique is sensitive to the fluorophore's local environment (temperature, ion concentration, dissolved gas concentration, and molecular associations), while being independent of factors impacting fluorescence intensity (fluorophore concentration, photobleaching, scattering, and absorption). We present design features of a novel UV-visible-NIR wide-field time-domain FLIM system with optical sectioning (10 μm), high temporal discrimination (50 ps), and large temporal dynamic range (750 ps - 1 μs), and apply the system to probe cellular metabolic function and detect molecular activity in vivo.

Citation Download Citation

Wei Zhong, Dhruv Sud, Mei Wu, Karl A. Merrick, Sofia D. Merajver, David G. Beer, and Mary-Ann Mycek "Sensing cellular function and molecular activity in vivo using fluorescence lifetime imaging microscopy (FLIM)", Proc. SPIE 5864, Novel Optical Instrumentation for Biomedical Applications II, 586407 (1 September 2005); https://doi.org/10.1117/12.632912

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