Experimental characterization of 3D localization techniques for particle-tracking and super-resolution microscopy

Mlodzianoski, Michael J.; Juette, Manuel F.; Beane, Glen L.; Bewersdorf, Joerg

doi:10.1364/oe.17.008264

Cited by 130 publications

(115 citation statements)

References 23 publications

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“…In a volume of 200 9 200 9 200 nm 3 around the focal point, the accuracy was reported to be B 26 nm for the in-plane, and B 52 nm for the out-of-plane position. Mlodzianoski et al (2009) compared the performance of bi-plane imaging and astigmatic imaging using a EMCCD camera. Both techniques showed a comparable measurement accuracy.…”

Section: Imaging Based On Aberrations (Astigmatism)mentioning

confidence: 99%

Particle imaging techniques for volumetric three-component (3D3C) velocity measurements in microfluidics

Cierpka

Kähler

2011

J Vis

131

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The reliable measurement of the 3D3C velocity field in microfluidic devices becomes more and more important for future optimization and developments for lab-on-a-chip applications or point-of-care medical diagnosis systems. In the past years, different particle-based imaging methods, such as confocal scanning microscopy, holography, stereoscopic and tomographic imaging or approaches based on defocused particle images or optical aberrations have been developed and applied successfully to measure velocity fields in microfluidic systems. The benefits and drawbacks of these methods will be discussed in detail as the proper understanding of the measurement principle is essential to select the most appropriate technique for a desired measurement application. Once an imaging method is chosen, the velocity can be estimated by correlation-based methods or tracking approaches. The advantages and disadvantages of both methods and the importance of image preprocessing will also be discussed in detail.

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Section: Imaging Based On Aberrations (Astigmatism)mentioning

confidence: 99%

Particle imaging techniques for volumetric three-component (3D3C) velocity measurements in microfluidics

Cierpka

Kähler

2011

J Vis

131

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“…Three-dimensional localizations are possible with SMACM techniques by breaking the axial symmetry of the standard point spread function (PSF) (14)(15)(16), imaging in multiple focal planes simultaneously (17), or projecting the axial dimension onto a lateral dimension (18). These methods can achieve precisions below 100 nm in 3D over a 2-μm range (14,19,20). In addition, sub-10-nm precision can be obtained via interferometry (21); however, these methods severely restrict the sample geometry, have a shallower operational depth of field (∼650 nm) (22), and are not yet conducive to live-cell imaging.…”

mentioning

confidence: 99%

Three-dimensional superresolution colocalization of intracellular protein superstructures and the cell surface in live Caulobacter crescentus

Lew

Lee

Ptacin

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

136

141

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Recently, single-molecule imaging and photocontrol have enabled superresolution optical microscopy of cellular structures beyond Abbe's diffraction limit, extending the frontier of noninvasive imaging of structures within living cells. However, livecell superresolution imaging has been challenged by the need to image three-dimensional (3D) structures relative to their biological context, such as the cellular membrane. We have developed a technique, termed superresolution by power-dependent active intermittency and points accumulation for imaging in nanoscale topography (SPRAIPAINT) that combines imaging of intracellular enhanced YFP (eYFP) fusions (SPRAI) with stochastic localization of the cell surface (PAINT) to image two different fluorophores sequentially with only one laser. Simple light-induced blinking of eYFP and collisional flux onto the cell surface by Nile red are used to achieve single-molecule localizations, without any antibody labeling, cell membrane permeabilization, or thiol-oxygen scavenger systems required. Here we demonstrate live-cell 3D superresolution imaging of Crescentin-eYFP, a cytoskeletal fluorescent protein fusion, colocalized with the surface of the bacterium Caulobacter crescentus using a double-helix point spread function microscope. Three-dimensional colocalization of intracellular protein structures and the cell surface with superresolution optical microscopy opens the door for the analysis of protein interactions in living cells with excellent precision (20-40 nm in 3D) over a large field of view (12 × 12 μm).cell biology | wide-field microscopy | cytoskeleton | fluorescence microscopy | bacteria

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“…Moreover, the defocusing patterns can be distorted by optical aberration when imaging deep regions. Experimental calibration of the PSF deviation is possible in some situations (29,30), but remains difficult in heterogeneous samples such as brain tissue.To overcome the challenges described above, we have developed a method, termed virtual volume superresolution microscopy (VVSRM), to map single fluorescent emitters in a virtual 3D space with near-isotropic 3D resolution. In this method, a tilted mirror is positioned near the sample region to generate a side view in addition to the front view.…”

mentioning

confidence: 99%

mentioning

confidence: 99%

Near-isotropic 3D optical nanoscopy with photon-limited chromophores

Tang

Akerboom

Vaziri

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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Imaging approaches based on single molecule localization break the diffraction barrier of conventional fluorescence microscopy, allowing for bioimaging with nanometer resolution. It remains a challenge, however, to precisely localize photon-limited single molecules in 3D. We have developed a new localization-based imaging technique achieving almost isotropic subdiffraction resolution in 3D. A tilted mirror is used to generate a side view in addition to the front view of activated single emitters, allowing their 3D localization to be precisely determined for superresolution imaging. Because both front and side views are in focus, this method is able to efficiently collect emitted photons. The technique is simple to implement on a commercial fluorescence microscope, and especially suitable for biological samples with photon-limited chromophores such as endogenously expressed photoactivatable fluorescent proteins. Moreover, this method is relatively resistant to optical aberration, as it requires only centroid determination for localization analysis. Here we demonstrate the application of this method to 3D imaging of bacterial protein distribution and neuron dendritic morphology with subdiffraction resolution.3D bioimaging | nanoimaging | bacterial imaging | neuron imaging | biophotonics F luorescence microscopy allows the observation of biological samples in a highly sensitive, selective, and relatively noninvasive fashion. Moreover, fluorescence imaging tools can probe deeply into tissues to obtain volume images without physically isolating different sections. However, traditional fluorescence microscopy suffers from diffraction-limited resolution, far larger than actual molecular sizes. Recent development of two different superresolution (SR) fluorescence imaging methods has broken the diffraction limit and enables direct optical observation of biological features near molecular scales. One approach is to create a subdiffraction excitation pattern by stimulated emission depletion (1, 2) or structured illumination (3, 4), and the other approach is based on single molecule localization (5-9). An essential part of the single molecule localization methods involves centroid fitting of serial single molecule images at the detection plane in order to determine the lateral localization of each fluorescent molecule with precision limited not by diffraction, but mainly by its number of emitted photons. A superresolution 2D image of the sample can then be rendered by overlapping all single molecule localizations from all image frames. This principle of localization has also been applied to single particle tracking (10)(11)(12)(13)(14).Single molecule-based superresolution microscopy requires individual chromophores to be turned "on" and "off" sequentially, either by photoswitching or by other stochastic photophysical processes. Compared to synthetic photoswitchable probes, photoactivatable fluorescent proteins (PA-FPs) (15-20) are superior in many regards: PA-FPs can be genetically fused to target proteins and endogenously...

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Experimental characterization of 3D localization techniques for particle-tracking and super-resolution microscopy

Cited by 130 publications

References 23 publications

Particle imaging techniques for volumetric three-component (3D3C) velocity measurements in microfluidics

Particle imaging techniques for volumetric three-component (3D3C) velocity measurements in microfluidics

Three-dimensional superresolution colocalization of intracellular protein superstructures and the cell surface in live Caulobacter crescentus

Near-isotropic 3D optical nanoscopy with photon-limited chromophores

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