Aberration-free optical refocusing in high numerical aperture microscopy

Botcherby, Edward; Juškaitis, Rimas; Booth, Martin J.; Wilson, Tony

doi:10.1364/ol.32.002007

Cited by 240 publications

(216 citation statements)

References 4 publications

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“…In the Fourier plane of an imaging system, the wavefront of light at wavelength λ, emanating from emitters located an axial distance of Δz away from the focal plane in a medium of index of refraction n λ , is characterized by a phase-shift profile that scales as ð2π=λÞn λ Δz (25). The distortion of the MFG acts on the emission wavefront to counterbalance this phase-shift profile (23).…”

Section: Resultsmentioning

confidence: 99%

Whole-cell, multicolor superresolution imaging using volumetric multifocus microscopy

Hajj

Wiśniewski

Beheiry

et al. 2014

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

102

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Single molecule-based superresolution imaging has become an essential tool in modern cell biology. Because of the limited depth of field of optical imaging systems, one of the major challenges in superresolution imaging resides in capturing the 3D nanoscale morphology of the whole cell. Despite many previous attempts to extend the application of photo-activated localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM) techniques into three dimensions, effective localization depths do not typically exceed 1.2 μm. Thus, 3D imaging of whole cells (or even large organelles) still demands sequential acquisition at different axial positions and, therefore, suffers from the combined effects of out-of-focus molecule activation (increased background) and bleaching (loss of detections). Here, we present the use of multifocus microscopy for volumetric multicolor superresolution imaging. By simultaneously imaging nine different focal planes, the multifocus microscope instantaneously captures the distribution of single molecules (either fluorescent proteins or synthetic dyes) throughout an ∼4-μm-deep volume, with lateral and axial localization precisions of ∼20 and 50 nm, respectively. The capabilities of multifocus microscopy to rapidly image the 3D organization of intracellular structures are illustrated by superresolution imaging of the mammalian mitochondrial network and yeast microtubules during cell division.superresolution | 3D localization | microscopy | multiplane imaging | single-molecule fluorescence

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Section: Resultsmentioning

confidence: 99%

Whole-cell, multicolor superresolution imaging using volumetric multifocus microscopy

Hajj

Wiśniewski

Beheiry

et al. 2014

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

102

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show abstract

“…It might be tempting to move the camera to a different imaging plane, but both this gives rise to spherical aberration and the images are much dimmer. Given that both the illumination and transmission objectives must remain stationary to obtain spectra, it was decided that instead the focal plane should be reimaged remotely (Botcherby et al 2007), which is achieved via microscope objectives MO 3 and MO 4 in Fig. 2.…”

Section: Spectral Selection Criteriamentioning

confidence: 99%

Towards a high-throughput real-time confocal microfluidic system for monitoring absorbance spectra in mixed-phase chemical reactions

Lawton

Girkin

2017

Microfluid Nanofluid

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“…In a first set of methods, refocusing was adapted [15,16] to oblique imaging. In the original version of refocusing, the Wilson group proposed to use two facing objectives (the first objective to refocus the object and the second objective to image the refocused object, or to use a single objective facing a mirror to achieve the same goal [15].…”

Section: Introductionmentioning

confidence: 99%

High-resolution real-time dual-view imaging with multiple point of view microscopy

Mangeol¹

2016

Biomed. Opt. Express

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Most methods to observe three-dimensional processes in living samples are based on imaging a single plane that is sequentially scanned through the sample. Sequential scanning is inherently slow, which can make it difficult to capture objects moving quickly in three dimensions. Here we present a novel method, multiple point-of-view microscopy (MPoVM), that allows simultaneous capturing of the front and side views of a sample with high resolution. MPoVM can be implemented in most fluorescence microscopes, offering new opportunities in the study of dynamic biological processes in three dimensions. 2. R. Tomer, K. Khairy, F. Amat, and P. J. Keller, "Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy," Nat. Methods 9(7), 755-763 (2012). 3. U. Krzic, S. Gunther, T. E. Saunders, S. J. Streichan, and L. Hufnagel, "Multiview light-sheet microscope for rapid in toto imaging," Nat. Methods 9(7), 730-733 (2012) imaging using single-objective SPIM," Nat. Methods 12(7), 641-644 (2015). 34. S.-I. Chang and J.-B. Yoon, "Shape-controlled, high fill-factor microlens arrays fabricated by a 3D diffuser lithography and plastic replication method," Opt. Express 12(25), 6366-6371 (2004).

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Aberration-free optical refocusing in high numerical aperture microscopy

Cited by 240 publications

References 4 publications

Whole-cell, multicolor superresolution imaging using volumetric multifocus microscopy

Whole-cell, multicolor superresolution imaging using volumetric multifocus microscopy

Towards a high-throughput real-time confocal microfluidic system for monitoring absorbance spectra in mixed-phase chemical reactions

High-resolution real-time dual-view imaging with multiple point of view microscopy

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