Improved axial resolution in confocal fluorescence microscopy using annular pupils

Gu, Miṅ; Tannous, Tony Abi; Sheppard, Colin J. R.

doi:10.1016/0030-4018(94)90245-3

Cited by 31 publications

(29 citation statements)

References 11 publications

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“…The images of the fluorescent layer were calculated by convolving a step function and the effective PSF. 46 The background rejection properties can be determined by the steepness of the edge response curve at the bottom (baseline response). In the calculation results ( Fig.…”

Section: Depth Imaging Property Of Sax Microscopymentioning

confidence: 99%

Saturated excitation microscopy for sub-diffraction-limited imaging of cell clusters

et al. 2013

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Saturated excitation (SAX) microscopy offers high-depth discrimination predominantly due to nonlinearity in the fluorescence response induced by the SAX. Calculation of the optical transfer functions and the edge responses for SAX microscopy revealed the contrast improvement of high-spatial frequency components in the sample structure and the effective reduction of background signals from the out-of-focus planes. Experimental observations of the edge response and x-z cross-sectional images of stained HeLa cells agreed well with theoretical investigations. We applied SAX microscopy to the imaging of three-dimensional cultured cell clusters and confirmed the resolution improvement at a depth of 40 μm. This study shows the potential of SAX microscopy for super-resolution imaging of deep parts of biological specimens.

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Section: Depth Imaging Property Of Sax Microscopymentioning

confidence: 99%

Saturated excitation microscopy for sub-diffraction-limited imaging of cell clusters

et al. 2013

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“…[65][66][67][68][69] Other geometries can also be used. For example, using annular apertures in conjunction with a confocal pinhole of¯nite size can also result in improved sectioning, 35,[70][71][72][73] and this approach can also be used with confocal°uorescence microscopy. 74,75 A rapidly developing approach is based on the illumination by a plane of light.…”

Section: Angular Gatingmentioning

confidence: 99%

“…For example, using annular apertures in conjunction with a confocal pinhole of¯nite size can also result in improved sectioning, 35,[70][71][72][73] and this approach can also be used with confocal°uorescence microscopy. 74,75 A rapidly developing approach is based on the illumination by a plane of light. These techniques are variously called light sheet microscopy, orthogonal-plane°uorescence optical sectioning (OPFOS), 76 selected plane illumination microscopy (SPIM), [77][78][79] or just ultra-microscopy 80 (named after the original paper by Siedentopf and Zsigmondy 75 ).…”

Section: Angular Gatingmentioning

confidence: 99%

“…74,75 A rapidly developing approach is based on the illumination by a plane of light. These techniques are variously called light sheet microscopy, orthogonal-plane°uorescence optical sectioning (OPFOS), 76 selected plane illumination microscopy (SPIM), [77][78][79] or just ultra-microscopy 80 (named after the original paper by Siedentopf and Zsigmondy 75 ). The light sheet can be generated using a scanned beam, and combined with the structured illumination technique.…”

Section: Angular Gatingmentioning

confidence: 99%

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Advanced optical microscopy methods for in vivo imaging of sub-cellular structures in thick biological tissues

Chen

Rehman

Sheppard

2014

J. Innov. Opt. Health Sci.

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Optical microscopy has become an indispensable tool for visualizing sub-cellular structures and biological processes. However, scattering in biological tissues is a major obstacle that prevents high-resolution images from being obtained from deep regions of tissue. We review common techniques, such as multiphoton microscopy (MPM) and optical coherence microscopy (OCM), for di®raction limited imaging beyond an imaging depth of 0.5 mm. Novel implementations have been emerging in recent years giving higher imaging speed, deeper penetration, and better image quality. Focal modulation microscopy (FMM) is a novel method that combines confocal spatial ltering with focal modulation to reject out-of-focus background. FMM has demonstrated an imaging depth comparable to those of MPM and OCM, near-real-time image acquisition, and the capability for multiple contrast mechanisms.

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“…19,20 This idea has been implemented in a large number of ways. [21][22][23][24][25][26][27] Many superresolution methods deal with the improvement of a single planar image. For 3D specimens (as is often the case in biological studies, for example), it is thus necessary to process each axial section separately, and sequentially, which can be time consuming.…”

Section: Introductionmentioning

confidence: 99%

Scanning holographic microscopy with resolution exceeding the Rayleigh limit of the objective by superposition of off-axis holograms

et al. 2007

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We present what we believe to be a new application of scanning holographic microscopy to superresolution. Spatial resolution exceeding the Rayleigh limit of the objective is obtained by digital coherent addition of the reconstructions of several off-axis Fresnel holograms. Superresolution by holographic superposition and synthetic aperture has a long history, which is briefly reviewed. The method is demonstrated experimentally by combining three off-axis holograms of fluorescent beads showing a transverse resolution gain of nearly a factor of 2.

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Improved axial resolution in confocal fluorescence microscopy using annular pupils

Cited by 31 publications

References 11 publications

Saturated excitation microscopy for sub-diffraction-limited imaging of cell clusters

Saturated excitation microscopy for sub-diffraction-limited imaging of cell clusters

Advanced optical microscopy methods for in vivo imaging of sub-cellular structures in thick biological tissues

Scanning holographic microscopy with resolution exceeding the Rayleigh limit of the objective by superposition of off-axis holograms

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