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Fluorescence lifetime imaging in an optically sectioning programmable array microscope (PAM).

Quentin S Hanley·Keith A Lidke·Rainer Heintzmann·Donna J Arndt-Jovin·Thomas M Jovin
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Abstract

BACKGROUND: The programmable array microscopes (PAMs) are a family of instruments incorporating arbitrary control of the patterns of illumination and/or detection. The PAM can be used in sectioning and nonsectioning modes, thereby constituting a useful platform for fluorescence lifetime imaging. METHODS AND RESULTS: We used a PAM for acquisition of optically sectioned and widefield fluorescence lifetime images, in which contrast was increased predominantly by suppressing out-of-focus light contributions. We simulate, display, and discuss the effects of blurring and fluorophore heterogeneity on lifetime imaging in widefield and confocal configurations. CONCLUSION: Sectioning improves the quality of lifetime images of samples with multiple fluorophores or spatially varying Förster resonance energy transfer.

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AnimalsDNA-Binding ProteinsDrosophila ProteinsEnergy TransferFluorescenceImage Processing, Computer-AssistedMicroscopyPolycomb Repressive Complex 1Journal Article

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Quentin S Hanley, Keith A Lidke, Rainer Heintzmann, Donna J Arndt-Jovin, Thomas M Jovin. 2005. Fluorescence lifetime imaging in an optically sectioning programmable array microscope (PAM).. https://doi.org/10.1002/cyto.a.20177

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