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检索条件"任意字段=Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXII"

共 590 条记录，以下是361-370 订阅

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Improved image-forming optics for transmission optical projection tomography

Improved image-forming optics for transmission optical proje...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： Wang, Yi Wang, Ruikang Oregon Hlth & Sci Univ Dept Biomed Engn Beaverton OR 97006 USA Foshan Univ Dept Photoelect & Phys Foshan 528000 Peoples R China

ISBN: (纸本)9780819465566

We demonstrate an improved image-forming optics for transmission optical projection tomography (TOPT), with which the parallel integral throughout an object can be obtained. This is performed by introducing a shutter with an appropriate diameter at the back focus of the objective lens. We evaluate and compare the performances of the improved and the conventional TOPT systems with different parameters to obtain the optimal configuration. The optimal reconstruction is achieved by the improved configuration with alpha = 0.8 degrees similar to 1.6 degrees, and the spatial resolution reaches 25 mu n. The optimal configuration is validated by TOPT of a phantom sample and a five days chick embryo.

关键词： optical projection tomography point spread function parallel integral

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Effects of specimen morphology on adaptive confocal and multiphoton microscopy

Effects of specimen morphology on adaptive confocal and mult...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： Booth, Martin J. Wilson, Tony Univ Oxford Dept Engn Sci Parks Rd Oxford OX1 3PJ England

ISBN: (纸本)9780819465566

microscopy of thick biological specimens is often detrimentally affected by specimen-induced aberrations. In the simplest case, these aberrations arise from a refractive index mismatch between the immersion and mounting media. In other situations, the aberrations arise from variations in refractive index within the specimen. These aberrations cause loss of signal and reduced resolution. Aberrations can be corrected using adaptive optics, where a deformable mirror introduces equal but opposite aberrations into the optical path. Aberration correction can be performed by reconfiguring the deformable mirror and using the fluorescence signal as feedback, effectively maximising the fluorescence intensity. However, the degree to which aberrations affect the intensity is related to the distribution of fluorescence in the specimen. Signals from point-like objects are affected more than equivalent signals from from planar or volume objects. We investigate this effect and discuss the implications for adaptive optical microscopy of biological specimens.

关键词： confocal microscopy multiphoton microscopy aberrations adaptive optics

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Confocal Fluorescence lifetime imaging microscopy based on a realtime sampling method

Confocal Fluorescence lifetime imaging microscopy based on a...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： Bae, Yoon. S. Lee, Dongsoo Moon, Sucbei Won, Young Jae Kim, Dug Y. Gwangju Inst Sci & Technol Dept Informat & Commun 1 Oryong Dong Kwangju 500712 South Korea

ISBN: (纸本)9780819465566

We present a fluorescence lifetime imaging microscope (FLIM) based on a real-time waveform acquisition method. The fluorophores were excited by a 635-mn gain-switched laser diode,, which produced short pulses with duration similar to 50 ps in a 20-MEz repetition rate. The fluorescence signals were detected by a silicon avalanche photo-diode (APD) in addition to a wide-band electric amplifier. The converted electric pulses were sampled by a high-speed digitizer of which sampling rate was 2 GS/s. In order to reduce the sampling interval for analyzing sub-nanosecond lifetimes, an interleaved data acquisition technique was used. The effective sampling rate was increased to 10 GS/s. In addition, the impulse response was measured simultaneously with the lifetime signals by an interleaving manner and was used in calibration of the system. By using these methods, accurate lifetime information was acquired in a short time less than 8 mu s.

关键词： confocal microscopy fluorescence lifetime imaging microscopy interleaved data acquisition

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microscopy using micro-pixellated light emitting diodes

Microscopy using micro-pixellated light emitting diodes

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： Poher, V. Kennedy, G. T. Elson, D. S. French, P. M. W. Neil, A. A. A. Imperial Coll London Blackett Lab South Kensington Campus London SW7 2AZ England

ISBN: (纸本)9780819465566

In this paper we demonstrate structured illumination microscopy, a cheap and flexible method of obtaining optical sectioning in wide-field microscopy, and we investigate line-scanning microscopy, both using the same microelement LED. In our first experiment we demonstrate structured illumination using a custom-designed microelement LED consisting of 120 individually addressable stripes emitting at 470 mn. An electrical driver was designed to produce a programmable grid pattern and the device was located in an Olympus BX41 fluorescence microscope in critical illumination configuration. By using an integrated solid-state alternative to a conventional piezo-actuated grid and separate illumination source, we improved the speed and accuracy of the system, reducing the artefacts due to the errors in the grid positions. Additionally, we investigated the use of the same LED device as a scanning source for confocal line-scanning microscopy. After each line scanning, an image was acquired using a CCD camera and the out-offocus light was rejected by a post-processing method.

关键词： structured illumination microscopy line scanning confocal microscopy micro-stripe light emitting diode

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Comparison of three dimensional transfer function analysis of alternative phase imaging methods

Comparison of three dimensional transfer function analysis o...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： Kou, Shan Shan Sheppard, Colin J. R. Natl Univ Singapore Div Bioengn 9 Engn Dr Singapore 117576 Singapore NUS Grad Sch Interact Sci & Engn Singapore 117597 Singapore

ISBN: (纸本)9780819465566

Phase imaging is an invaluable tool for observation of biological sample, especially for living cells, where staining might not be appropriate, or for materials that do not absorb stain. Imaging of phase distributions with high spatial resolution can be used to derive the actual thickness and refractive index variations in the specimen. The detection of very small phase variations enables the detailed structure in the specimen to be revealed. As a result, the development and utilization of various phase imaging modalities have been important aspects of microscopy research. Differential Interference Contrast (DIC) and Quantitative Phase Microcopy (QPM) are based on partially coherent light, thus enabling high-resolution imaging. However, the low coherence requirement prevents the acquisition of quantitative phase data directly. On the other hand, Digital Holography microscopy (DHM) is able to yield quantitative phase information but is compromised on resolution and cannot give full three dimensional (3D) reconstructions. In this paper, we present the 3D theoretical formalism of the above mentioned phase imaging methods with the focus on DHM. A comparative analysis here through visualization of 3D optical transfer functions gives an insight into the behaviors of these phase imaging methods.

关键词： phase imaging 3D optical transfer function

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High-resolution laser scanning microscopy with saturated excitation of fluorescence

High-resolution laser scanning microscopy with saturated exc...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： Fujita, Katsumasa Kawano, Shogo Kobayashi, Minoru Kawata, Satoshi Osaka Univ Dept Appl Phys 2-2 Yamadaoka Suita Osaka 5650871 Japan Osaka Univ Dept Frontier Biosci Suita Osaka 5650871 Japan Nanophoton Corp Suita Osaka 5650871 Japan RIKEN Wako Saitama 3510198 Japan

ISBN: (纸本)9780819465566

We propose the use of saturated excitation to improve the spatial resolution of a laser scanning confocal fluorescence microscope. The saturated excitation induces nonlinear response in fluorescence emission that gives higher-order spatial frequency components in the point spread fuction of the microscope. To extract the nonlinear responses in fluorescence emission, we modulate the excitation intensity temporally at a single frequency (omega) and demodulate the fluorescence signal at the harmonic frequencies (2 omega, 3 omega,...). We found that the fluorescence signal demodulated at nth harmonic frequency is proportional to nth power of the excitation intensity, where n-fold improvement of the spatial resolution can be achieved. We experimentally confirmed that the demodulated signal at the second harmonic frequency was proportional to the square of the excitation intensity. The improvement of spatial resolution was also confirmed by observing a fluorescent sample.

关键词： laser scanning microscopy confocal microscopy fluorescence saturated excitation high-resolution microscopy

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Calibration of a phase-shifting DIC microscope for quantitative phase imaging

Calibration of a phase-shifting DIC microscope for quantitat...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： King, Sharon V. Libertun, Ariel R. Preza, Chrysanthe Cogswell, Carol J. Univ Colorado Dept Elect & Comp Engn Boulder CO 80309 USA Univ Memphis Dept Elect & Syst Engn Memphis TN USA CDM Opt Inc Boulder CO USA

ISBN: (纸本)9780819465566

Phase-shifting differential interference contrast (DIC) provides images in which the intensity of DIC is transformed into values linearly proportional to differential phase delay. Linear regression analysis of the Fourier space, spiral phase, integration technique shows these values can be integrated and calibrated to provide accurate phase measurements of objects embedded in optically transparent media regardless of symmetry or absorption properties. This approach has the potential to overcome the limitations of profilometery, which cannot access embedded objects, and extend the capabilities of the traditional DIC microscope, which images embedded phase objects, but does not provide quantitative information.

关键词： DIC (Nomarski) microscopy quantitative phase imaging phase shifting spiral phase

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Development of a random access multi-photon microscope for fast three-dimensional functional recording of neuronal activity

Development of a random access multi-photon microscope for f...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： Reddy, Duemani Saggau, Peter Rice Univ Dept Bioengn Houston TX 77251 USA Baylor Coll Med Dept Neurosci Houston TX 77030 USA

ISBN: (纸本)9780819465566

Over the past two decades, the dendritic processes of neurons have been shown to possess active and dynamic properties that give them the ability to modulate synaptic integration and shape individual synaptic responses. Effectively studying these properties at multiple locations on a live neuron in highly scattering brain. tissue requires an imaging/recording mechanism with high spatiotemporal resolution as well as optical sectioning and random access site selection capabilities. Our lab has made significant steps in developing such a system by combining the spatial resolution and optical sectioning ability of imaging techniques such as confocal and multi-photon microscopy with the temporal resolution and random access capability provided by acousto-optic laser scanning. However, all systems that have been developed to date restrict fast imaging to two-dimensional (2D) scan patterns. This severely limits the extent to which many neurons can be studied since they represent complex three-dimensional (3D) structures. We have previously demonstrated a scheme for fast 3D scanning which utilizes a unique arrangement of multiple acousto-optic deflectors and does not require axial movements of the objective lens. Here we couple this scanning scheme to a modified commercial research microscope and use the combined system to effectively image user-defined sites of interest on fluorescent 3D structures with positioning times that are in the low microsecond range. The resulting random-access scanning mechanism allows for functional imaging of complex 3D cellular structures such as neuronal dendrites at frames rates on the order of tens of kilohertz.

关键词： Neurons 3D scanning Microscopes 3D image processing Imaging systems Objectives Tissue optics Acousto-optics Confocal microscopy Laser scanners

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Investigation of image properties in super-resolution microscopy using two-color fluorescence dip spectroscopy

Investigation of image properties in super-resolution micros...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： Iketaki, Yoshinori Watanabe, Takeshi Bokor, Nandor Fujii, Masaaki Olympus Corp Adv Core Technol Dept 2-3 Kuboyama Cho Hachioji Tokyo 1928512 Japan Budapest Univ Technol Econ Dept Phys H-1111 Budapest Hungary Tokyo Inst Technol Chem Resources Lab Midori Ku Yokohama Kanagawa 2268503 Japan

ISBN: (纸本)9780819465566

We quantitatively investigated image properties in super-resolution microscopy using two-color fluorescence dip spectroscopy. To evaluate the properties, the point spread function (PSF) and contrast transfer fuNction (CTF) were measured using a fluorescent scale together with a fluorescent bead. From the CTF, it has been found that visible light can resolve a 100 mn line-and-space pattern by microcopy, and provide a contrast of 10%. The CTF corresponds to a PSF with a FWHM of 130 mn. The value is two times finer than the diffraction limit size. An evaluation using a 100 run 0 fluorescent bead consistently supports the result given by the CTF for super-resolution microscopy. The measured CTF shows that super-resolution microscopy can indeed improve the optical properties of fluorescent images and enable us to observe a structure with a spatial resolution overcoming the diffraction limit.

关键词： two-color dip spectroscopy contrast transfer function point spread function fluorescence-depletion process super-resolution microscopy Rhodamine 6G

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Extension of multidimensional microscopy to ultra-sensitive applications with maximum-likelihood analysis

Extension of multidimensional microscopy to ultra-sensitive ...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： Davis, Lloyd M. Shen, Guoqing Univ Tennessee Inst Space Ctr Laser Applicat Tullahoma TN 37388 USA

ISBN: (纸本)9780819465566

multidimensional fluorescence microscopy is finding service in forefront biological studies that require separation of images from different fluorophores. For example, commercial microscopes are available with multi-band analog detectors and user-friendly software for "linear unmixing" of species with overlapping emission spectra. To extend such techniques to ultrasensitive and single-molecule applications, we have developed a custom-built microscope, which incorporates two tunable-wavelength picosecond dye lasers for pulse-interleaved laser excitation, angle-tuned reflection of the laser beams from narrow-band Raman notch filters to introduce epi-illumination and provide strong rejection of scattered laser wavelengths, diffraction-limited confocal imaging with 3-dimensional piezo-scanning, an adjustable prism spectrometer for high-throughput resolution of collected fluorescence into 4 spectral bands, and a 4-channel high-quantum efficiency avalanche diode for sub-nanosecond-resolved single-photon detection. Custom software enables multi-band fluorescence correlation spectroscopy and identification of photon bursts for single-molecule detection. For umnixing of spectrally-overlapping signatures for ultrasensitive molecular imaging applications, we find that maximum-likelihood analysis can out-perform least-squares-based linear unmixing in the regime of low photon numbers per spectral/temporal channel. Also, the likelihood surface provides the confidence of the parameter estimates and the covariance of the species contributions. Monte Carlo simulations show that bias in the results of the analysis, which stems from the constraint that photon numbers should be positive, becomes more pronounced at low signal levels, for both maximum-likelihood and least-squares based uninixing.

关键词： fluorescence microscopy spectral-FLIM maximum-likelihood multidimensional microscopy photon counting

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