Three-dimensional phase contrast imaging by an annular illumination microscope

Noda, Tomoya; Kawata, Satoshi; Minami, Shigeo

doi:10.1364/ao.29.003810

Cited by 27 publications

(28 citation statements)

References 5 publications

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“…Eq. 2 is not difficult to evaluate numerically, however, and the paraxial 3D phase OTF may be replaced by a non-paraxial version as in [21] for high numerical aperture imaging. Figure 1a, for example, shows the PCTF's numerically calculated for two defocus distances ( = ±0.6 and ±9 ) and ̅ = 546 .…”

Section: Principles Of Bright-field Quantitative Phase Microscopymentioning

confidence: 99%

Bright-field quantitative phase microscopy (BFQPM) for accurate phase imaging using conventional microscopy hardware

Jenkins¹,

Gaylord²

2015

SPIE Proceedings

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Most quantitative phase microscopy methods require the use of custom-built or modified microscopic configurations which are not typically available to most bio/pathologists. There are, however, phase retrieval algorithms which utilize defocused bright-field images as input data and are therefore implementable in existing laboratory environments. Among these, deterministic methods such as those based on inverting the transport-of-intensity equation (TIE) or a phase contrast transfer function (PCTF) are particularly attractive due to their compatibility with Köhler illuminated systems and numerical simplicity. Recently, a new method has been proposed, called multi-filter phase imaging with partially coherent light (MFPI-PC), which alleviates the inherent noise/resolution trade-off in solving the TIE by utilizing a large number of defocused bright-field images spaced equally about the focal plane. Despite greatly improving the state-ofthe-art, the method has many shortcomings including the impracticality of high-speed acquisition, inefficient sampling, and attenuated response at high frequencies due to aperture effects. In this report, we present a new method, called bright-field quantitative phase microscopy (BFQPM), which efficiently utilizes a small number of defocused bright-field images and recovers frequencies out to the partially coherent diffraction limit. The method is based on a noiseminimized inversion of a PCTF derived for each finite defocus distance. We present simulation results which indicate nanoscale optical path length sensitivity and improved performance over MFPI-PC. We also provide experimental results imaging live bovine mesenchymal stem cells at sub-second temporal resolution. In all, BFQPM enables fast and accurate phase imaging with unprecedented spatial resolution using widely available bright-field microscopy hardware.

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Section: Principles Of Bright-field Quantitative Phase Microscopymentioning

confidence: 99%

Bright-field quantitative phase microscopy (BFQPM) for accurate phase imaging using conventional microscopy hardware

Jenkins¹,

Gaylord²

2015

SPIE Proceedings

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“…[6][7][8] While the purpose of each procedure is the same, i.e. acquiring data along the surface of the Ewald sphere, it was shown recently that the two approaches lead to different transfer functions, and therefore different resolutions along each axis.…”

Section: Tomography Principlementioning

confidence: 99%

Optical tomography by digital holographic microscopy

et al. 2009

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Optical tomography provides three-dimensional data of the measured specimen, while quantitative phase imaging enables measuring the induced phase-shifts. Combining those two technologies makes possible to get three-dimensional refractive index reconstruction. This can be achieved by introducing a scan in the measurement process, which can be done in several ways. We present and compare results of tomographic measurements, taken either in angle-scanning or wavelength-scanning approach, respectively in transmission or in reflection microscopy, in the framework of digital holographic microscopy.

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“…Indeed, the 3-D OTF has been calculated for a laser-scan fluorescence microscope [11]. An annular illumination microscope has also been constructed and investigated recently [12]. However, due to the low value of C being used, the gain in depth-of-focus was not significant enough and depth scanning was required for 3-D inspection [12].…”

Section: Annular Pupil and Its Long Dept-of-focusmentioning

confidence: 99%

“…An annular illumination microscope has also been constructed and investigated recently [12]. However, due to the low value of C being used, the gain in depth-of-focus was not significant enough and depth scanning was required for 3-D inspection [12]. However, the use of an annular aperture for C 1 has a major drawback in that it stops and wastes a large amount of light and, therefore, in particularly, it has not been used effectively for fluorescence microscopes for the purpose of achieving larger depth of focus for 3-D microscopic imaging.…”

Section: Annular Pupil and Its Long Dept-of-focusmentioning

confidence: 99%

Difference-of-Gaussian annular pupil for extended depth of focus three-dimensional imaging

Poon

Kourakos

2000

Optical Sensing, Imaging, and Manipulation for Biological and Biomedical Applications

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We propose a technique for 3-D microscopic imaging with extended depth-of-focus using a novel illumination scheme in a laser scanning optical microscope. The novel illumination scheme creates an effective annular pupil, called the difference-of-Gaussian annular pupil, without the critical drawback of stopping and wasting the light. Two laser beams of different Gaussian pupils with different temporal frequencies are first generated. The laser beams are then combined spatially and used to scan the specimen. The scattered light from the object is picked up by a photodetector whose output consists of a DC and an AC current (due to the optical heterodyning of the two optical beams). The DC signal is no difference from the DC output of a conventional laser scanning microscope with the processing pupil as a Gaussian function, whereas the AC signal is derived from the mixing of the two Gaussian beams and would be given by effectively a Gaussian pupil with a different size than that generated by the DC signal. The AC and the DC signals are then subtracted by electronics and hence the effective pupil function would be given by the difference of the two Gaussian pupil functions. By properly choosing the size of the two Gaussian laser beams, we could realize the difference of the two Gaussian pupils which becomes a new type of annular pupil called the difference-of-Gaussian annular pupil.

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Three-dimensional phase contrast imaging by an annular illumination microscope

Cited by 27 publications

References 5 publications

Bright-field quantitative phase microscopy (BFQPM) for accurate phase imaging using conventional microscopy hardware

Bright-field quantitative phase microscopy (BFQPM) for accurate phase imaging using conventional microscopy hardware

Optical tomography by digital holographic microscopy

Difference-of-Gaussian annular pupil for extended depth of focus three-dimensional imaging

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