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Super-resolution localization microscopy with photoactivatable fluorescent marker proteins

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

Fluorescent proteins (FPs) have become popular imaging tools because of their high specificity, minimal invasive labeling and allowing visualization of proteins and structures inside living organisms. FPs are genetically encoded and expressed in living cells, therefore, labeling involves minimal effort in comparison to approaches involving synthetic dyes. Photoactivatable FPs (paFPs) comprise a subclass of FPs that can change their absorption/emission properties such as brightness and color upon irradiation. This methodology has found a broad range of applications in the life sciences, especially in localization-based super-resolution microscopy of cells, tissues and even entire organisms. In this review, we discuss recent developments and applications of paFPs in super-resolution localization imaging.

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Abbreviations

AFM:

Atomic force microscopy

DNA:

Deoxyribonucleic acid

FRET:

Förster resonance energy transfer

FP:

Fluorescent protein

FPALM:

Fluorescence photoactivation localization microscopy

GFP:

Green fluorescent protein

MRI:

Magnetic resonance imaging

OCT:

Optical coherence tomography

paFP:

Photoactivatable fluorescent protein

PALM:

Photoactivated localization microscopy

PET:

Positron emission tomography

PSF:

Point spread function

RESOLFT:

Reversible saturable optical fluorescence transitions

SOFI:

Stochastic optical fluctuation imaging

SPIM:

Selective plane illumination microscopy

SSIM:

Saturated structured illumination microscopy

STED:

Stimulated emission depletion

STORM:

Stochastic optical reconstruction microscopy

TIRF:

Total internal reflection fluorescence

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Acknowledgments

This work was supported by the Deutsche Forschungsgemeinschaft (DFG) and the State of Baden-Württemberg through the Center for Functional Nanostructures (CFN) and by DFG grant Ni 291/9.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Applied Physics, Karlsruhe Institute of Technology (KIT), 76128, Karlsruhe, Germany

    Per Niklas Hedde & G. Ulrich Nienhaus

  2. Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, CA, 92697, USA

    Per Niklas Hedde

  3. Institute of Toxicology und Genetics, Karlsruhe Institute of Technology (KIT), 76021, Karlsruhe, Germany

    G. Ulrich Nienhaus

  4. Department of Physics, University of Illinois at Urbana–Champaign, Urbana, IL, 61801, USA

    G. Ulrich Nienhaus

Authors
  1. Per Niklas Hedde
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  2. G. Ulrich Nienhaus
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Correspondence to G. Ulrich Nienhaus.

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Handling Editor: J. W. Borst

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Hedde, P.N., Nienhaus, G.U. Super-resolution localization microscopy with photoactivatable fluorescent marker proteins. Protoplasma 251, 349–362 (2014). https://doi.org/10.1007/s00709-013-0566-z

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  • DOI: https://doi.org/10.1007/s00709-013-0566-z

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