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Imaging of Lanthanide Luminescence by Time-Resolved Microscopy

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Book cover Lanthanide Luminescence

Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 7))

Abstract

Time-resolved fluorescence microscopy (TRFM) of objects stained with luminescent lanthanides chelate-based reagents requires adaptation of microscope equipment to distinguish the long-lived fluorescence of the lanthanide from scattered excitation light and relatively short-lived autofluorescence of biological objects and optical components. How this is accomplished depends on the choice of image scanning, e.g., wide-field microscopy (image plane scanning) or object plane scanning [(confocal) laser microscopy]. Typically, excitation light is presented as pulses by applying mechanically chopping of a continuous light beam, or by using flash lamps or pulse lasers. The detector (CCD camera, photomultiplier tube) is time-gated to record the delayed signal in a defined time interval, either electronically or using phase-locked choppers in the emission pathway. This chapter discusses strategies for optimal TRFM and reviews the main existing configurations used for lanthanide imaging. In addition, special techniques such are Fluorescence Lifetime Imaging Microscopy (FLIM) and Time-Correlated Single Photon Counting (TCSPC) are presented and discussed as well.

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Abbreviations

CCD:

Charge-coupled device

DELFIA:

Dissociation enhanced lanthanide fluoroimmunoassay

FLIM:

Fluorescence lifetime imaging microscopy

FRET:

Fluorescence resonance energy transfer

LED:

Light emitting diode

MRI:

Magnetic resonance imaging

PMT:

Photomultiplier tube

TCSPC:

Time-correlated single photon counting

TR-FIA:

Time-resolved fluoroimmuno assay

TRFM:

Time-resolved fluorescence microscopy

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Authors and Affiliations

  1. Department of Molecular Cell Biology, Leiden University Medical Centre, Einthovenweg 20, Bldg 2, 9600, 2300 RC, Leiden, The Netherlands

    H. J. Tanke

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  1. H. J. Tanke
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Correspondence to H. J. Tanke .

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Editors and Affiliations

  1. , Department of Cell Biology and Anatomy, University of Turki, Tykistökatu 6A, Turku, 20520, Finland

    Pekka Hänninen

  2. , Department of Cell Biology and Anatomy, University of Turku, Tykistökatu 6A, Turku, 20520, Finland

    Harri Härmä

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Tanke, H.J. (2010). Imaging of Lanthanide Luminescence by Time-Resolved Microscopy. In: Hänninen, P., Härmä, H. (eds) Lanthanide Luminescence. Springer Series on Fluorescence, vol 7. Springer, Berlin, Heidelberg. https://doi.org/10.1007/4243_2010_2

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