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

TY - JOUR

T1 - Fluorescence lifetime imaging in an optically sectioning programmable array microscope (PAM)

AU - Hanley, Q S

AU - Lidke, K A

AU - Heintzmann, R

AU - Arndt-Jovin, D J

AU - Jovin, T M

PY - 2005/10

Y1 - 2005/10

N2 - 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 Forster resonance energy transfer. (c) 2005 International Society for Analytical Cytology

AB - 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 Forster resonance energy transfer. (c) 2005 International Society for Analytical Cytology

U2 - 10.1002/cyto.a.20177

DO - 10.1002/cyto.a.20177

M3 - Article

VL - 67A

SP - 112

EP - 118

JO - CYTOMETRY PART A

JF - CYTOMETRY PART A

IS - 2

ER -