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Fluorescence nanoscopy. Methods and applications

  • Review
  • Published:
Journal of Chemical Biology

Abstract

Fluorescence nanoscopy refers to the experimental techniques and analytical methods used for fluorescence imaging at a resolution higher than conventional, diffraction-limited, microscopy. This review explains the concepts behind fluorescence nanoscopy and focuses on the latest and promising developments in acquisition techniques, labelling strategies to obtain highly detailed super-resolved images and in the quantitative methods to extract meaningful information from them.

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Notes

  1. Please note that structured illumination microscopy is also commonly used to designate a diffraction-limited optical sectioning methodology based on patterned illumination [98].

Abbreviations

3B:

Bayesian analysis of the bleaching and blinking

βME:

β-Mercaptoethanol

φFl :

Fluorescence quantum yield

AChR:

Acetylcholine receptor

BaLM:

Bleaching/blinking assisted localisation microscopy

CHO:

Chinese hamster ovary

CW:

Continuous wave

dSTORM:

Direct STORM

EM:

Electron microscopy

ER:

Endoplasmic reticulum

EMCCD:

Electron-multiplying CCD

FCS:

Fluorescence correlation spectroscopy

FIONA:

Fluorescence imaging with 1 nm accuracy

FlAsH:

Fluorescein arsenical helix

FP:

Fluorescent protein

FPALM:

Fluorescent photoactivation light microscopy

FWHM:

Full width half maximum

GPI:

Glycophosphatidylinositol

GSH:

Glutathione

GSD:

Ground state depletion

GSDIM:

Ground state depletion followed by individual molecule return

GPU:

Graphics processing unit

HA:

Hemagglutinin

HIV-1:

Human immunodeficiency virus type 1

LM:

Localisation microscopy

MEA:

Mercaptoethylamine

MLE:

Maximum likelihood estimate

NA:

Numerical aperture

NK:

Natural killer

NPC:

Nuclear pore complex

PA-FP:

Photoactivatable fluorescent protein, it is usually referred to any photoactivatable, photoswitchable or photochromic FP

PAINT:

Paint accumulation for imaging in nanoscale topography

PALM:

Photoactivation localisation microscopy

PALMIRA:

Photoactivation light microscopy with independent running acquisition

PIP2:

Phosphatidylinositol 4,5-bisphosphate

PSF:

Point spread function

RESOLFT:

Reversible, saturable, optical fluorescence transition

SIM:

Structured illumination microscopy

SOFI:

Stochastic optical fluctuation imaging

SSIM:

Saturated structured illumination

STORM:

Stochastic optical reconstruction microscopy

STED:

Stimulated emission depletion

TCR:

T cell antigen receptor

TfR:

Transferrin receptor

TIRF:

Total internal reflection fluorescence

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Acknowledgments

The author gratefully acknowledges funding from the Ministerio de Ciencia e Innovación of Spain (MICINN Plan Nacional contract number FIS2009-07966) and the continuing support of the Fundación Biofísica-Bizkaia, in Spain. The author also thanks Gloria de las Heras and Unai Lorenzo for helpful comments and critical revision of the manuscript and Asier Ruiz (Achucarro Basque Centre for Neurosciences) for permission to reproduce Fig. 3.

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  1. Unidad de Biofísica, CSIC-UPV/EHU, Barrio de Sarriena, s/n., 48940, Leioa, Spain

    Jose Requejo-Isidro

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Requejo-Isidro, J. Fluorescence nanoscopy. Methods and applications. J Chem Biol 6, 97–120 (2013). https://doi.org/10.1007/s12154-013-0096-3

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