Multiphoton imaging and fluorescence lifetime studies on unstained cells and tissue at cryogenic conditions

Martin Stark; Daniel Dörr; Alexander Ehlers; Daniel Sauer; Rainer Bückle; Sven Martin; Friederike Ehrhart; Jennifer Baunach; Alisa Katsen-Globa; Heiko Zimmermann; Karsten König

doi:10.1117/12.728317

11 July 2007 Multiphoton imaging and fluorescence lifetime studies on unstained cells and tissue at cryogenic conditions

Martin Stark, Daniel Dörr, Alexander Ehlers, Daniel Sauer, Rainer Bückle, Sven Martin, Friederike Ehrhart, Jennifer Baunach, Alisa Katsen-Globa, Heiko Zimmermann, Karsten König

Author Affiliations +

Proceedings Volume 6628, Diagnostic Optical Spectroscopy in Biomedicine IV; 662809 (2007) https://doi.org/10.1117/12.728317
Event: European Conferences on Biomedical Optics, 2007, Munich, Germany

Abstract

Monitoring the functional status of cryo-preserved cells and tissue in-situ, i.e. in the frozen state, might allow for optimal adjustment of preservation conditions and might provide the information necessary to predict a functionality recovery rate. Here, an imaging approach with compositional sensitivity seems favourable. In our approach we use multiphoton microscopy in combination with fluorescence lifetime imaging to investigate cells, human and plant tissue at cryogenic conditions. By the non-linearity of multiphoton excitation we largely suppress image distortions attributed to scattering of incoming light. Only where the intensity of the pulsed near-infrared laser beam is sufficiently large, significant fluorescence is excited. This allows reaching penetration depth in ice comparable to the liquid state. As additional information we use the fluorescence decay to assign compositional entities. Results obtained on cells and tissues are discussed with respect to temperature dependencies and the related use for applications.

Citation Download Citation

Martin Stark, Daniel Dörr, Alexander Ehlers, Daniel Sauer, Rainer Bückle, Sven Martin, Friederike Ehrhart, Jennifer Baunach, Alisa Katsen-Globa, Heiko Zimmermann, and Karsten König "Multiphoton imaging and fluorescence lifetime studies on unstained cells and tissue at cryogenic conditions", Proc. SPIE 6628, Diagnostic Optical Spectroscopy in Biomedicine IV, 662809 (11 July 2007); https://doi.org/10.1117/12.728317

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