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Fast fluorescence holographic microscopy

Fast fluorescence holographic microscopy

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： Qin, Wan Yang, Xiaoqi Li, Yingying Peng, Xiang Qu, Xinghua Yao, Hai Gao, Bruce Z. Clemson Univ Dept Bioengn Clemson SC 29634 USA Shenzhen Univ Inst Optoelect Shenzhen 518060 Peoples R China Tianjin Univ State Key Lab Precis Measuring Technol & Instrum Tianjin 300072 Peoples R China

ISBN: (纸本)9780819498625

FINCHSCOPE is a new technology of fluorescence holographic microscopy. It has been successfully applied to recording high-resolution three-dimensional fluorescence images of biological specimens without the need for scanning. In this study, we revealed and analyzed an intrinsic phenomenon, called ghost lens effect, on spatial light modulator which is the core element enabling the incoherent correlation in the FINCHSCOPE. The ghost lens effect can degrade the imaging quality by introducing multiple spherical waves with different focal lengths into the correlation and thus increasing the noise in the recorded holograms.

关键词： Fluorescence holography spatial light modulator FINCHSCOPE ghost lens effect

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Real-time stroboscopic full-field optical coherence tomography based on graphics processing unit

Real-time stroboscopic full-field optical coherence tomograp...

引用

conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： Park, Kwan Seob Kim, Hyounggyu Kim, Ju Wan Baek, Hee Gyu Lee, Jae Hwi Chung, Youngjoo Lee, Byeong Ha Gwangju Inst Sci & Technol Sch Informat & Commun 261 Cheomdan Gwagiro Kwangju 500712 South Korea Gwangju Inst Sci & Technol Dept Med Syst Engn Gwangju 500712 South Korea

ISBN: (纸本)9780819498625

We present the real-time stroboscopic full-field optical coherence tomography (FF-OCT) system that is based on graphics processing unit (GPU). The basic configuration of the proposed FF-OCT system was the Linnik interferometer. While scanning of a reference mirror in the axial direction, a series of the transverse sectional image was captured with a 2-dimensional CCD camera. To get a depth-resolved 3-D image, the light source of OCT was turned on and off like a stroboscope at the Doppler frequency of the OCT system. The CCD camera used in experiment operated at a rate of 200 frames per second, but the Doppler frequency was similar to kHz. To overcome the slow operation of the CCD camera below the Doppler frequency, the light source was operated in the stroboscopic mode. In addition, lock-in detection technique was utilized in order to avoid the dissolution of the coherent signals during the integration time of the CCD camera. Furthermore, the Doppler frequency shift due to nonlinear scanning motion of the reference mirror was monitored by using an auxiliary interferometer and then fed back to the light source driver so that the strobe frequency was always matched with the Doppler frequency of the OCT system. For the real-time 3-D rendering, we used a graphics processing unit.

关键词： full-field optical coherence tomography stroboscopic illumination graphics processing unit real-time rendering

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Flash extreme ultraviolet holographic microscopy in a table top setup

Flash extreme ultraviolet holographic microscopy in a table ...

引用

conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： Monserud, Nils C. Malm, Erik B. Wachulak, Przemyslaw W. Xu, Huiwen Balakrishnan, Ganesh Chao, Weilun Anderson, Erik Marconi, Mario C. Colorado State Univ Engn Res Ctr Extreme Ultraviolet Sci & Technol Dept Elect & Comp Engn Ft Collins CO 80523 USA Mil Univ Technol Inst Optoelctron PL-00908 Warsaw Poland Univ New Mexico Ctr High Technol Mat Dept Elect & Comp Engn Albuquerque NM 87106 USA Univ Calif Berkeley Lawrence Berkeley Natl Lab Ctr Xray Opt Berkeley CA 94720 USA

ISBN: (纸本)9780819498625

We present a single-shot Fourier transform holography setup with similar to 100nm spatial resolution and 1 ns temporal resolution using a tabletop extreme ultraviolet (EUV) laser. Flash images allowed for the imaging of nano-pillars oscillating at MHz frequencies that will enable the evaluation of mechanical properties of nanoscale mechanical oscillators.

关键词： Holography Extreme ultraviolet Holograms microscopy Zone plates Fourier transforms Scanning electron microscopy Oscillators Spatial resolution Temporal resolution

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Reflective interferometric system combining low-coherence spectral-domain phase microscopy and wide-field holography for characterization of thin samples

Reflective interferometric system combining low-coherence sp...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： Friedman, Reut Shaked, Natan T. Tel Aviv Univ Fac Engn Dept Biomed Engn IL-69978 Tel Aviv Israel

ISBN: (纸本)9780819498625

We introduce an integrated system combining low-coherence spectral-domain phase microscopy (SDPM) together with a compact, simple-to-align, wide-field high-coherence interferometer (off-axis TAU module) for accurate quantitative phase measurements. The proposed compact system is capable of capturing an off-axis, wide-field interference in the time domain simultaneously with single-point interference in the spectral domain. The integrated system can obtain both quantitative phase of transparent samples, requiring a reflective surface at the back of the sample, or profiling of reflective samples. Since there are no moving elements in the system, it is capable of measuring static and dynamic samples, while time resolution is limited only by the frame rate of the detectors (a camera and a compact spectrometer). Both interferometers are in common-path geometry, resulting in high signal to noise ratio and nanometer-scale stability. The combined system is ideal for characterization of static or dynamic samples containing both wide areas of interest that can be acquired by the off-axis TAU module, and specific spots of interest requiring fine measurement that can be acquired by SDPM

关键词： Digital holography Holographic interferometry microscopy Interference microscopy phase microscopy low-coherence interferometry optical coherence tomography Hybrid interferometric microscope Reflective interferometry

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Wavefront shaping of a Bessel light field enhances light sheet microscopy with scattered light

Wavefront shaping of a Bessel light field enhances light she...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： Nylk, J. Mitchell, C. Vettenburg, T. Gunn-Moore, F. J. Dholakia, K. Univ St Andrews Sch Phys & Astron SUPA St Andrews KY16 9SS Fife Scotland Univ St Andrews Sch Biol St Andrews KY16 9TF Fife Scotland

ISBN: (纸本)9780819498625

Light sheet microscopy has seen a resurgence as it facilitates rapid, high contrast, volumetric imaging with minimal sample exposure. Initially developed for imaging scattered light, this application of light sheet microscopy has largely been overlooked but provides an endogenous contrast mechanism which can complement fluorescence imaging and requires very little or no modification to an existing light sheet fluorescence microscope. Fluorescence imaging and scattered light imaging differ in terms of image formation. In the former the detected light is incoherent and weak whereas in the latter the coherence properties of the illumination source, typically a laser, dictate the coherence of detected light, but both are dependent on the quality of the illuminating light sheet. image formation in both schemes can be understood as the convolution of the light sheet with the specimen distribution. In this paper we explore wavefront shaping for the enhancement of light sheet microscopy with scattered light. We show experimental verification of this result, demonstrating the use of the propagation invariant Bessel beam to extend the field of view of a high resolution scattered light, light sheet microscope and its application to imaging of biological super-cellular structures with sub-cellular resolution. Additionally, complementary scattering and fluorescence imaging is used to characterize the enhancement, and to develop a deeper understanding of the differences of image formation between contrast mechanisms in light sheet microscopy.

关键词： Light sheet microscopy LSM selective plane illumination microscopy SPIM beam shaping Bessel beam high resolution deconvolution

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in vivo Stepwise Multi-Photon Activation Fluorescence Imaging of Melanin in Human Skin

<i>in vivo</i> Stepwise Multi-Photon Activation Fluorescence...

引用

conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： Lai, Zhenhua Gu, Zetong Abbas, Saleh Lowe, Jared Sierra, Heidy Rajadhyaksha, Milind DiMarzio, Charles Northeastern Univ Dept Elect & Comp Engn Boston MA 02115 USA Northeastern Univ Biol Dept Boston MA 02115 USA Northeastern Univ Mech & Ind Engn Dept Boston MA 02115 USA Mem & Sloan Kettering Canc Ctr Dermatol Serv New York NY 10065 USA Bernard M Gordon Ctr Subsurface Sensing & Imagin San Juan PR 00931 USA

ISBN: (纸本)9780819498625

The stepwise multi-photon activated fluorescence (SMPAF) of melanin is a low cost and reliable method of detecting melanin because the activation and excitation can be a continuous-wave (CW) mode near infrared (NIR) laser. Our previous work has demonstrated the melanin SMPAF images in sepia melanin, mouse hair, and mouse skin. In this study, we show the feasibility of using SMPAF to detect melanin in vivo. in vivo melanin SMPAF images of normal skin and benign nevus are demonstrated. SMPAF images add specificity for melanin detection than MPFM images and CRM images. Melanin SMPAF is a promising technology to enable early detection of melanoma for dermatologists.

关键词： fluorescence stepwise multi-photon excitation melanin microscopy near-infrared

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Quantitative Phase microscopy: how to make phase data meaningful

Quantitative Phase Microscopy: how to make phase data meanin...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： Goldstein, Goldie Creath, Katherine 4D Technol Corp Tucson AZ 85706 USA

ISBN: (纸本)9780819498625

The continued development of hardware and associated image processing techniques for quantitative phase microscopy has allowed superior phase data to be acquired that readily shows dynamic optical volume changes and enables particle tracking. Recent efforts have focused on tying phase data and associated metrics to cell morphology. One challenge in measuring biological objects using interferometrically obtained phase information is achieving consistent phase unwrapping and background shape removal throughout a sequence of images. Work has been done to improve the phase unwrapping in two- dimensions and correct for temporal discrepanices using a temporal unwrapping procedure. The residual background shape due to mean value fluctuations and residual tilts can be removed automatically using a simple object characterization algorithm. Once the phase data are processed consistently, it is then possible to characterize biological samples such as myocytes and myoblasts in terms of their size, texture and optical volume and track those features dynamically. By observing optical volume dynamically it is possible to determine the presence of objects such as vesicles within myoblasts even when they are co-located with other objects. Quantitative phase microscopy provides a label-free mechanism to characterize living cells and their morphology in dynamic environments, however it is critical to connect the measured phase to important biological function for this measurement modality to prove useful to a broader scientific community. In order to do so, results must be highly consistent and require little to no user manipulation to achieve high quality nynerical results that can be combined with other imaging modalities.

关键词： phase imaging interference microscopy polarization interferometry cellular imaging cell dynamics optical thickness measurement

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Selective plane illumination microscopy with structured illumination based on spatial light modulators

Selective plane illumination microscopy with structured illu...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： Li, Runze Zhou, Xing Wu, Di Peng, Tong Yang, Yanlong Lei, Ming Yu, Xianhua Yao, Baoli Ye, Tong Chinese Acad Sci Xian Inst Opt & Precis Mech State Key Lab Transient Opt & Photon Xian Shaanxi Peoples R China Clemson Univ Ctr Opt Mat Sci & Engn Technol Dept Bioengn Clemson SC 29631 USA

ISBN: (纸本)9780819498625

Structured-illumination microscopy (SIM) is an efficacious tool to decrease the contribution of the out-of-focus light to images of specimens. However, in SIM, the frequency of the spatial modulation applied to specimens should be adjustable according to the optical properties of the specimens to reach the optimal contrasts. Hence, a common theme in SIM is how the flexibility and quality of modulations at different frequencies can be improved. Digital scanned laser light-sheet microscopy with structured illumination (DSLM-SI) has been the most flexible means for generating modulation and optical sectioning. The complexity of synchronization between the temporal modulation and the beam scanning makes it hard to use and less stable;it also takes more time to acquire images for one plane than selective plane illumination microscopy (SPIM). In this report, we present a recent effort to use a spatial light modulator (SLM) to provide spatial modulation in SPIM. With the SLM, both of the frequency and phase of lateral modulation can be changed rapidly;moreover, this SLM-based SPIM can achieve fast imaging without mechanical moving parts.

关键词： Structured-illumination light-sheet spatial light modulator digital scanned laser light-sheet single-plane illumination

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Portable advanced off-axis camera for quantitative phase microscopy

Portable advanced off-axis camera for quantitative phase mic...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： Monemhaghdoust, Z. Depeursinge, C. Moser, C. Ecole Polytech Fed Lausanne Lab Appl Photon Devices CH-1015 Lausanne Switzerland Lyncee Tec SA CH-1015 Lausanne Switzerland

ISBN: (纸本)9780819498625

We propose and experimentally demonstrate a device in which common-path interferometry combined with off-axis holographic geometry is used to realize a digital holographic camera which can be attached to the camera port of a conventional transmission microscope for complex wavefront analysis. A thick transmission volume grating recorded holographically into thick photosensitive glass splits the beam containing the sample information in two beams. The untouched transmitted beam creates the sample arm of the interferometer. The Bragg diffracted order of the grating is spectrally and spatially filtered by diffraction to generate a clean reference beam. Double passing the diffracted order through the grating using a retroreflector device provides filtering in two dimensions. The spatial filtering done by the grating which works based on high angular selectivity of thick volume gratings, reduces the alignment spatial sensitivity which is an advantage over the conventional spatial filtering done by pinholes. Besides, using a second thick grating, we introduce a desired coherence plane tilt in the reference beam which is sufficient to create high-visibility interference over the entire field of view. The full-field off-axis interferograms are created from which the amplitude and phase can be reconstructed. The advantage of the proposed camera is the insensitivity to the alignment, thus can be the basis for a standalone camera mountable on a standard optical microscope.

关键词： Diffraction gratings Digital holographic microscopy Portable camera Off-axis full-field imaging Phase microscopy

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Implementation of PSF Engineering in High-Resolution 3D microscopy Imaging with a LCoS (Reflective) SLM

Implementation of PSF Engineering in High-Resolution 3D Micr...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： King, Sharon V. Doblas, Ana Patwary, Nurmohammed Saavedra, Genaro Mart-Nez-Corral, Manuel Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA Univ Valencia Dept Opt 3D Imaging & Display Lab E-46100 Burjassot Spain

ISBN: (纸本)9780819498625

Wavefront coding techniques are currently used to engineer unique point spread functions (PSFs) that enhance existing microscope modalities or create new ones. Previous work in this field demonstrated that simulated intensity PSFs encoded with a generalized cubic phase mask (GCPM) are invariant to spherical aberration or misfocus;dependent on parameter selection. Additional work demonstrated that simulated PSFs encoded with a squared cubic phase mask (SQUBIC) produce a depth invariant focal spot for application in confocal scanning microscopy. Implementation of PSF engineering theory with a liquid crystal on silicon (LCoS) spatial light modulator (SLM) enables validation of WFC phase mask designs and parameters by manipulating optical wavefront properties with a programmable diffractive element. To validate and investigate parameters of the GCPM and SQUBIC WFC masks, we implemented PSF engineering in an upright microscope modified with a dual camera port and a LCoS SLM. We present measured WFC PSFs and compare them to simulated PSFs through analysis of their effect on the microscope imaging system properties. Experimentally acquired PSFs show the same intensity distribution as simulation for the GCPM phase mask, the SQUBIC-mask and the well-known and characterized cubic-phase mask (CPM), first applied to high NA microscopy by Arnison et al.(10), for extending depth of field. These measurements provide experimental validation of new WFC masks and demonstrate the use of the LCoS SLM as a WFC design tool. Although efficiency improvements are needed, this application of LCoS technology renders the microscope capable of switching among multiple WFC modes.

关键词： microscopy wave-front encoding spatial light modulators liquid crystal

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