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FRET-FLIM applications in plant systems

  • Special Issue: New/Emerging Techniques in Biological Microscopy
  • Published:
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Abstract

A hallmark of cellular processes is the spatio-temporally regulated interplay of biochemical components. Assessing spatial information of molecular interactions within living cells is difficult using traditional biochemical methods. Developments in green fluorescent protein technology in combination with advances in fluorescence microscopy have revolutionised this field of research by providing the genetic tools to investigate the spatio-temporal dynamics of biomolecules in live cells. In particular, fluorescence lifetime imaging microscopy (FLIM) has become an inevitable technique for spatially resolving cellular processes and physical interactions of cellular components in real time based on the detection of Förster resonance energy transfer (FRET). In this review, we provide a theoretical background of FLIM as well as FRET-FLIM analysis. Furthermore, we show two cases in which advanced microscopy applications revealed many new insights of cellular processes in living plant cells as well as in whole plants.

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Abbreviations

ACR4:

Arabidopsis crinkly 4

BiFC:

Bimolecular fluorescence complementation

BRI1:

Brassinosteroid insensitive 1

CLV1:

Clavata1

CPK21:

Calcium-dependent protein kinase 21

FCS:

Fluorescence correlation spectroscopy

FCCS:

Fluorescence cross-correlation spectroscopy

FLIM:

Fluorescence lifetime imaging microscopy

FRET:

Förster resonance energy transfer

FLS2:

Flagellin sensing 2

GFP:

Green fluorescent protein

LRR-RLK:

Leucine-rich repeat receptor-like kinases

MFIS:

Multiparameter fluorescence image spectroscopy

PM:

Plasma membrane

SERK:

Somatic embryogenesis receptor-like kinase

STED:

Stimulated emission depletion

TCSPC:

Time-correlated single photon counting

TIRF:

Total internal reflection fluorescence

VFP:

Visible fluorescent protein

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. The Sainsbury Laboratory, Norwich Research Park, Norwich, NR4 7UH, UK

    Christoph A. Bücherl

  2. Laboratory of Biophysics and Microspectroscopy Centre, Wageningen University, Wageningen, The Netherlands

    Arjen Bader

  3. School of Chemistry, University of East Anglia, Norwich, NR4 7TJ, UK

    Sergey P. Laptenok

  4. Laboratory of Biochemistry and Microspectroscopy Centre, Wageningen University, Wageningen, The Netherlands

    Adrie H. Westphal & Jan Willem Borst

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  1. Christoph A. Bücherl
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  2. Arjen Bader
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  3. Adrie H. Westphal
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  4. Sergey P. Laptenok
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  5. Jan Willem Borst
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Correspondence to Jan Willem Borst.

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Bücherl, C.A., Bader, A., Westphal, A.H. et al. FRET-FLIM applications in plant systems. Protoplasma 251, 383–394 (2014). https://doi.org/10.1007/s00709-013-0595-7

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