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Picosecond Fluorescence Lifetime Imaging Spectroscopy as a New Tool for 3D Structure Determination of Macromolecules in Living Cells

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New Trends in Fluorescence Spectroscopy

Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 1))

Abstract

Picosecond fluorescence lifetime imaging microscopy (Picosecond FLIM), based on time-and space-correlated single photon counting (TSCSPC), is the method of choice to study living cells at minimal-invasive conditions that are required for preserving the living state Time-resolved fluorescence imaging, at ultra-low excitation intensity and ultra-low level of labelling, i e., Minimal-Invasive Fluorescence Microscopy (MIFM), became possible after the introduction of ultra-sensitive single photon counting imaging detectors, such as micro-channel plate (MCP) photomultipliers (PMT) with delay-line (DL) and quadrant anode (QA) The novel detectors have a time resolution of < 10 ps, 100 urn space resolution (250 × 250 channel), a dynamic range of >107 and are capable of Vehicle Micro-Spectroscopy (VMS) Singlechannel time-correlated single photon counting (TCSPC) microscopy was established 15 years ago and has since been applied by a growing community of cell biologists, due to superior performance and increasingly simple operation of pulsed picosecond laser systems The new TSCSPC imaging detectors will further increase the attractiveness of the well-established method in ultra-sensitive studies of living cells.

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Authors
  1. K. Kemnitz
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Editors and Affiliations

  1. Conservatoire National des Arts et Métiers, Laboratoire de Chimie Général, 292 rue Saint-Martin, 75141, Paris Cedex 03, France

    Bernard Valeur

  2. Ecole Normale Supérieure de Cachan, 61 Av. du Président Wilson, 94235, Cachan Cedex, France

    Jean-Claude Brochon

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© 2001 Springer-Verlag Berlin Heidelberg

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Kemnitz, K. (2001). Picosecond Fluorescence Lifetime Imaging Spectroscopy as a New Tool for 3D Structure Determination of Macromolecules in Living Cells. In: Valeur, B., Brochon, JC. (eds) New Trends in Fluorescence Spectroscopy. Springer Series on Fluorescence, vol 1. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-56853-4_18

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  • DOI: https://doi.org/10.1007/978-3-642-56853-4_18

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-63214-3

  • Online ISBN: 978-3-642-56853-4

  • eBook Packages: Springer Book Archive

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