Super-resolution Microscopy

Thorley, Jennifer; Pike, Jeremy; Rappoport, Joshua Z.

doi:10.1016/b978-0-12-409513-7.00014-2

Cited by 37 publications

(20 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Over the last couple of decades, ways to circumvent the Rayleigh limit in far-field fluorescence microscopy have been invented [2]. Confocal methods such as STED, RESOLFT, and SSIM [3][4][5][6] use patterned illumination to spatially modulate the fluorescence pattern of emitters within a diffraction-limited region such that not all of them emit simultaneously, thereby achieving sub-Rayleigh resolution.…”

Section: Introductionmentioning

confidence: 99%

Quantum limits of localisation microscopy

2019

View full text Add to dashboard Cite

Localisation microscopy of multiple weak, incoherent point sources with possibly different intensities in one spatial dimension is equivalent to estimating the amplitudes of a classical mixture of coherent states of a simple harmonic oscillator. This enables us to bound the multi-parameter covariance matrix for an unbiased estimator for the locations in terms of the quantum Fisher information matrix, which we obtained analytically. In the regime of arbitrarily small separations we find it to be no more than rank two-implying that no more than two independent parameters can be estimated irrespective of the number of point sources. We use the eigenvalues of the classical and quantum Fisher information matrices to compare the performance of spatial-mode demultiplexing and direct imaging in localisation microscopy with respect to the quantum limits.

show abstract

Section: Introductionmentioning

confidence: 99%

Quantum limits of localisation microscopy

2019

View full text Add to dashboard Cite

show abstract

“…In the last twenty years different techniques were developed to overcome this diffraction limit. The most commonly used super-resolution techniques are STED, SIM and SMLM all of which have been commercialized in the last few years (for detailed reviews of the different techniques see123). …”

mentioning

confidence: 99%

“…Theoretical discussions of the individual strengths of super-resolution techniques have been published12326 but a practical comparison of the performance of all three techniques has been missing to date. Here we imaged the same kinds of samples with SIM, STED and SMLM.…”

mentioning

confidence: 99%

Imaging cellular structures in super-resolution with SIM, STED and Localisation Microscopy: A practical comparison

Wegel

Göhler

Lagerholm

et al. 2016

Sci Rep

178

135

View full text Add to dashboard Cite

Many biological questions require fluorescence microscopy with a resolution beyond the diffraction limit of light. Super-resolution methods such as Structured Illumination Microscopy (SIM), STimulated Emission Depletion (STED) microscopy and Single Molecule Localisation Microscopy (SMLM) enable an increase in image resolution beyond the classical diffraction-limit. Here, we compare the individual strengths and weaknesses of each technique by imaging a variety of different subcellular structures in fixed cells. We chose examples ranging from well separated vesicles to densely packed three dimensional filaments. We used quantitative and correlative analyses to assess the performance of SIM, STED and SMLM with the aim of establishing a rough guideline regarding the suitability for typical applications and to highlight pitfalls associated with the different techniques.

show abstract

“…The resolution of this approach is dependent on the spot size of the remaining excited molecules, and resolutions of 29-60 nm have been achieved with this approach [9,10]. It is very important to make sure the depletion laser wavelength does not overlap with the excitation wavelength of the dye, or the resolution will be degraded significantly [11]. In addition, the type of dye and specimen mounting medium can both affect image quality [12].…”

Section: Review Of Super-resolution Microscopymentioning

confidence: 99%

Detectors for Super-Resolution & Single-Molecule Fluorescence Microscopies

Youker¹

2018

Photon Counting - Fundamentals and Applications

View full text Add to dashboard Cite

The resolution of light microscopy was thought to be limited to 250-300 nanometers based on the work of Ernest Abbe. This Abbe diffraction limit was believed to be insurmountable until the invention of Super-resolution microscopic techniques in the late 20th century. These techniques remove this limit and have provided unprecedented detail of cellular structures and dynamics down to several nanometers. An emerging goal in this field is to quantitatively measure individual molecules. Measurement of single-molecule dynamics, such as diffusion coefficients and complex stoichiometries, can be accomplished using fluorescence fluctuation techniques to reveal nanosecond-to-microsecond temporal reactions. These powerful complimentary experimental approaches are made possible by sensitive low-light photodetectors. In this chapter, an overview of the principles of super-resolution and single-molecule microscopies are provided. The different types of photodetectors employed in these techniques are explained. In addition, the advantages and disadvantages for these detectors are discussed, as well as the development of next generation detectors. Finally, example super-resolution and single-molecule cellular studies that take advantage of these detector technologies are presented.

show abstract

Super-resolution Microscopy

Cited by 37 publications

References 40 publications

Quantum limits of localisation microscopy

Quantum limits of localisation microscopy

Imaging cellular structures in super-resolution with SIM, STED and Localisation Microscopy: A practical comparison

Detectors for Super-Resolution & Single-Molecule Fluorescence Microscopies

Contact Info

Product

Resources

About