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检索条件"任意字段=Conference on 3-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX"

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Volumetric retinal fluorescence microscopic imaging with extended depth of field 13

Volumetric retinal fluorescence microscopic imaging with ext...

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

作者： Li, Zengzhuo Fischer, Andrew Li, Wei Li, Guoqiang Ohio State Univ Dept Ophthalmol & Visual Sci Columbus OH 43212 USA Ohio State Univ Dept Elect & Comp Engn Columbus OH 43212 USA Ohio State Univ Dept Neurosci Columbus OH 43212 USA NEI Bethesda MD 20892 USA

ISBN: (纸本)9781628419474

Wavefront-engineered microscope with greatly extended depth of field (EDoF) is designed and demonstrated for volumetric imaging with near-diffraction limited optical performance. A bright field infinity-corrected transmissive/reflective light microscope is built with Kohler illumination. A home-made phase mask is placed in between the objective lens and the tube lens for ease of use. General polynomial function is adopted in the design of the phase plate for robustness and custom merit function is used in Zemax for optimization. The resulting EDoF system achieves an engineered point spread function (PSF) that is much less sensitive to object depth variation than conventional systems and therefore 3D volumetric information can be acquired in a single frame with expanded tolerance of defocus. In Zemax simulation for a setup using 32X objective (NA = 0.6), the EDoF is 20 mu m whereas a conventional one has a DoF of 1.5 mu m, indicating a 13 times increase. In experiment, a 20X objective lens with NA = 0.4 was used and the corresponding phase plate was designed and fabricated. Retinal fluorescence images of the EDoF microscope using passive adaptive optical phase element illustrate a DoF around 100 mu m and it is able to recover the volumetric fluorescence images that are almost identical to in-focus images after post processing. The image obtained from the EDoF microscope is also better in resolution and contrast, and the retinal structure is better defined. Hence, due to its high tolerance of defocus and fine restored image quality, EDoF optical systems have promising potential in consumer portable medical imaging devices where user's ability to achieve focus is not optimal, and other medical imaging equipment where achieving best focus is not a necessary.

关键词： microscopy 3D imaging extended depth of field point spread function wavefront engineering passive adaptive optics eye imaging fluorescence imaging

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Interferometric computed-microtomography of 3D phase objects

Interferometric computed-microtomography of 3D phase objects

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conference on Three-dimensional microscopy - image acquisition and processing IV

作者： Vishnyakov, GN Levin, GG Zakarian, CS Russian Research Institute for Optophysical Measurements (Russia) Moscow State Technical Univ. (Russia)

ISBN: (纸本)0819423955

Measurements of refractive index spatial distribution within optically transparent phase 3-D samples with computerized interferomertric microscopes is proposed. For the reconstruction of the phase information the phase shifting interferometry is used, Interferometric computed-tomography microscope are fully automated integrated systems that incorporate CCD camera, frame grabber, computed operated PZT mirror as well as intelligible software for interferogram processing, tomographic reconstruction and refractive index distribution are displayed in 2-D and 3-D, sections, isolines, etc. Experimental results for interferometric microscopy and tomography of blood cells are presented.

关键词： computed tomography 3D microscopy interferometry phase object phase-shifting method refractive index erythrocyte

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A novel method for image denoising of fluorescence molecular imaging based on fuzzy C-Means clustering 22

A novel method for image denoising of fluorescence molecular...

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

作者： An, Yu Liu, Jie Ye, Jinzuo Mao, Yamin Yang, Xin Jiang, Shixin Chi, Chongwei Tian, Jie Beijing Jiaotong Univ Sch Comp & Informat Technol Dept Biomed Engn Beijing 100044 Peoples R China Inst Chinese Acad Sci Key Lab Mol Imaging Beijing 100190 Peoples R China

ISBN: (纸本)9781628414202

As an important molecular imaging modality, fluorescence molecular imaging (FMI) has the advantages of high sensitivity, low cost and ease of use. By labeling the regions of interest with fluorophore, FMI can noninvasively obtain the distribution of fluorophore in-vivo. However, due to the fact that the spectrum of fluorescence is in the section of the visible light range, there are mass of autofluorescence on the surface of the bio-tissues, which is a major disturbing factor in FMI. Meanwhile, the high-level of dark current for charge-coupled device (CCD) camera and other influencing factor can also produce a lot of background noise. In this paper, a novel method for image denoising of FMI based on fuzzy C-Means clustering (FCM) is proposed, because the fluorescent signal is the major component of the fluorescence images, and the intensity of autofluorescence and other background signals is relatively lower than the fluorescence signal. First, the fluorescence image is smoothed by sliding-neighborhood operations to initially eliminate the noise. Then, the wavelet transform (WLT) is performed on the fluorescence images to obtain the major component of the fluorescent signals. After that, the FCM method is adopt to separate the major component and background of the fluorescence images. Finally, the proposed method was validated using the original data obtained by in vivo implanted fluorophore experiment, and the results show that our proposed method can effectively obtain the fluorescence signal while eliminate the background noise, which could increase the quality of fluorescence images.

关键词： image denoising fluorescence molecular imaging fuzzy C-Means clustering

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Signal post processing in frequency domain OCT and OCM using a filter bank approach

Signal post processing in frequency domain OCT and OCM using...

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

作者： Hofer, Bernd Povazay, Boris Hermann, Boris Unterhuber, Angelika Matz, Gerald Hlawatsch, Franz Drexler, Wolfgang Cardiff Univ Dept Optometry & Vis Sci Biomed Imaging Grp Cardiff Wales Vienna Univ Technol Inst Commun & Radio Frequency Engn Vienna Austria

ISBN: (纸本)9780819465566

Current signal post processing in spectrally encoded frequency domain (FD) optical coherence microscopy (OCM) and optical coherence tomography (OCT) uses Fourier transforms in combination with non-uniform resampling strategies to map the k-space data acquired by the spectrometer to spatial domain signals which are necessary for tomogram generation. We propose to use a filter bank (FB) framework for the remapping process. With our new approach, the spectrometer is modeled as a critically sampled analysis FB, whose outputs are quantized subband signals that constitute the k-space spectroscopic data. The optimal procedure to map this data to the spatial domain is via a suitably designed synthesis FB which has low complexity. FB theory additionally states that 1) it is possible to find a synthesis FB such that the overall system has the perfect reconstruction (PR) property;2) any processing on critically sampled subband signals (as done in current schemes) results in aliasing artifacts. These perspectives are evaluated both theoretically and experimentally. We determine the analysis FB corresponding to our FD-OCM system by using a tunable laser and show that for our grating-based spectrometer - employing a CCD-line camera - the non-uniform resampling together with FFT indeed causes aliasing terms and depth dependent signal attenuation. Furthermore, we compute a finite impulse response based synthesis FB and assess the desired PR property by means of layered samples. The resulting images exhibit higher resolution and improved SNR compared to the common FFT-based approach. The potential of the proposed FB approach opens a new perspective also for other spectroscopic applications.

关键词： filter bank spectrometer spectroscopy optical coherence microscopy optical coherence tomography frequency domain tunable lasers fourier transform aliasing FFT

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Practical optical quality assessment and correction of a nonlinear microscope

Practical optical quality assessment and correction of a non...

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

作者： Aviles-Espinosa, Rodrigo Andilla, Jordi Porcar-Guezenec, Rafael Olarte, Omar Santos, Susana I. C. O. Levecq, Xavier Artigas, David Loza-Alvarez, Pablo ICFO Inst Photon Sci Mediterranean Technol PkAv Canal Olimp S-N Castelldefels 08860 Barcelona Spain Imagine Opt F-91400 Orsay France Univ Politecn Cataluna Barcelona 08034 Spain

ISBN: (纸本)9780819479662

Nonlinear microscopy (NLM) has covered the requirement for higher contrast and resolution compared with other microscopy techniques, however, the optical quality of this imaging apparatus and the sample structure can compromise its capabilities. Here, we show that the imaging capabilities of a NLM can be affected by the aberrations produced by the setup optical elements alignment, the materials from which they are fabricated and more importantly by the sample. To overcome this, a Shack-Hartman Wavefront sensing scheme has been implemented for characterizing: a) the whole NLM setup and, b) the sample induced aberrations. The first part includes all the aberrations introduced by the optical elements, starting from the laser and until the microscope objective. Having these information, aberrations can be compensated in a closed-loop configuration resulting in the system calibration. Then the remaining aberrations (microscope objective and sample) are recorded. This is done employing the sample nonlinear fluorescence signal collected at one point (keeping the excitation beam static) in the imaging plane. Given that this emission is an incoherent process, it can be considered as a point source. Therefore its wavefront will contain the sample and the objective aberrations. Using the wavefront sensor the information is recorded and passed to the deformable mirror which will compensate the aberrations in a "single shot" (open-loop configuration). This compared with other adaptive optics strategies (i.e. iterative algorithms) results in a reduced sample exposure, and greatly decreases sample damage. Importantly the application of both corrections (system and sample) enables a significant signal intensity and contrast improvement.

关键词： Adaptive Optics Nonlinear microscopy Aberration compensation

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Confocal laser microscopy of the in vivo human skin revisited.

Confocal laser microscopy of the <i>in vivo</i> human skin r...

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conference on 3-dimensional and multidimensional microscopy - image acquisition and processing VI

作者： Corcuff, P Hadjur, C Chaussepied, C Toledo-Crow, R LOreal Labs Rech Avancee F-93601 Aulnay Sous Bois France

ISBN: (纸本)0819430757

A new generation of confocal laser microscope, designed to image the human skin in vivo, improves the resolution, contrast and spectroscopic facilities as compared to the previous Tandem Scanning Microscope (TSM) prototype. The new device has been built with a Oz module (Noran) equipped with the skin contact device, assuming a perfect stability of skin images in the horizontal plane. The Z displacement of the objective lens, mounted directly on the Ch module, is assumed by a piezo motor with a course of 350 mu m. Moreover, the Ch module has been suspended on articulated arms to reach any part of the human body. The power of the Argon/Krypton laser source has been limited to 2 mW to secure safety and provides three visible wavelength : 488, 568 and 647 nm. The facility of instantly checking wavelength during in depth exploration of the skin optimizes the resolution and contrast of images as compared with the white light used in the TSM. Consequently, better image quality of the epidermis is obtained in the blue region with unexpected details of corneocytes and keratinocytes. The papillary dermis comprising the vascular network is advantageously observed with the red light. The fluorescence channel detector gives additional information on the penetration of fluorescent probes through the skin barrier. Optical sections are digitized (512 x 480 x 8 bit) at video rate, providing easy and fast measurements of the thickness of epidermal layers. The Silicon Graphics workstation generates a transparent volume of living human skin in less than 5 minutes. This powerful and convivial new design for imaging the in vivo human skin opens up new promises in skin research and in vivo skin pharmacology.

关键词： confocal microscopy human skin in vivo imaging fluorescence volume rendering

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Signal evaluation in chromatic confocal spectral interferometry via k-space phase equality approach 26

Signal evaluation in chromatic confocal spectral interferome...

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

作者： Claus, D. Boettcher, T. Ding, L. Taphanel, M. Osten, W. Inst Lasertechnol Med & Messtech Helmholtzstr 12 D-89081 Ulm Germany Inst Tech Opt Pfaffenwaldring 9 D-70569 Stuttgart Germany Fraunhofer Inst Optron Syst Tech & Bildauswertung Fraunhoferstr 1 D-76131 Karlsruhe Germany

ISBN: (纸本)9781510624092

Chromatic confocal spectral interferometry combines the benefits of scanning free acquisition of the axial dimension with interferometrically increased depth accuracy. However, so far it has been difficult to separate the confocal signal from the interferometric signal. It is, of course, possible to apply the established CCM evaluation methods. In that case, the available phase information, that offers a decreased measurement uncertainty and to some degree the removal of disturbing artifacts at steep surface inclinations, is not taken into account. In fact, it is not straight forward to interpret the signal. In comparison to white light interference microscopy, the signal suffers from a chirp. This means that it cannot be associated with a single beating frequency, which corresponds to the interferometrically encoded z-value. However, a modified lock-in technique has in the past successfully been applied, demonstrating a significant advantage in comparison to the conventional CCM procedures. Here, we will introduce the concept of k-space phase equality, which enables the separation of the confocal and the interferometric signal and furthermore offers an extended measurement range. The principle is based on signal modification in the z-space, which corresponds to the Fourier domain of the recorded spectral signal. The evaluation is then performed in the spectral domain, where the phase signals for all z-positions with respect to the corresponding wavelength are evaluated. As a result, a phase signal with reduced aberration terms, similar to an interferometric signal, is obtained, which can hence be evaluated using established techniques.

关键词： Chromatic confocal spectral interferometry chromatic confocal microscopy confocal microscopy signal evaluation

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Three-dimensional microscopy: image acquisition and processing II

Three-Dimensional Microscopy: Image Acquisition and Processi...

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Three-dimensional microscopy: image acquisition and processing II

ISBN: (纸本)0819417599

This conference proceedings contain 28 papers. Topics covered different aspects related to image acquisition and processing in 3D microscopy including: Biological and medical microscopy;Optical devices and components;Imaging systems;Mathematical techniques in processing;Calibration;and, Performance of a variety of techniques like confocal microscopy, laser scanning microscopy, holographic microscopy, wide-field microscopy, x-ray microtomography, and, scanning electron stereo microscopy.

关键词： Optical microscopy

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Imaging a seizure model in zebrafish with structured illumination light sheet microscopy 25

Imaging a seizure model in zebrafish with structured illumin...

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conference on Three-dimensional and multidimensional microscopy: image acquisition and processing XXV

作者： Liu, Yang Dale, Savannah Ball, Rebecca VanLeuven, Ariel J. Baraban, Scott Sornborger, Andrew Lauderdale, James D. Kner, Peter Univ Georgia Coll Engn Athens GA 30602 USA Clemson Univ Dept Bioengn Clemson SC 29634 USA Univ Georgia Dept Cellular Biol Athens GA 30602 USA Univ Calif San Francisco Dept Neurol Surg San Francisco CA 94143 USA Univ Calif Davis Dept Math Davis CA 95616 USA

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

Zebrafish are a promising vertebrate model for elucidating how neural circuits generate behavior under normal and pathological conditions. The Baraban group first demonstrated that zebrafish larvae are valuable for investigating seizure events and can be used as a model for epilepsy in humans. Because of their small size and transparency, zebrafish embryos are ideal for imaging seizure activity using calcium indicators. Light-sheet microscopy is well suited to capturing neural activity in zebrafish because it is capable of optical sectioning, high frame rates, and low excitation intensities. We describe work in our lab to use light-sheet microscopy for high-speed long-time imaging of neural activity in wildtype and mutant zebrafish to better understand the connectivity and activity of inhibitory neural networks when GABAergic signaling is altered in vivo. We show that, with light-sheet microscopy, neural activity can be recorded at 23 frames per second in two-colors for over 10 minutes allowing us to capture rare seizure events in mutants. We have further implemented structured illumination to increase resolution and contrast in the vertical and axial directions during high-speed imaging at an effective frame rate of over 7 frames per second.

关键词： fluorescence microscopy light sheet microscopy structured illumination microscopy neurophotonics

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Quantification of collagen networks in mammary tumors using TPEF and laser-based tomography 29

Quantification of collagen networks in mammary tumors using ...

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

作者： Kamin, Hannes Nolte, Lena Maurer, Jochen Bleilevens, Andreas Stickeler, Elmar Johannsmeier, Sonja Heinemann, Dag Ripken, Tammo Laser Zentrum Hannover eV Ind & Biomed Opt Dept Hollerithallee 8 D-30419 Hannover Germany Univ Hosp Aachen Clin Gynecol & Obstet Pauwelstr 30 D-52074 Aachen Germany Lower Saxony Ctr Biomed Engn Implant Res & Dev NI Stadtfelddamm 34 D-30625 Hannover Germany Leibniz Univ Hannover Inst Hort Prod Syst Dept Phytophoton Herrenhauser Str 2 D-30419 Hannover Germany Leibniz Univ Hannover Hannover Ctr Opt Technol HOT Nienburger Str 17 D-30167 Hannover Germany

ISBN: (纸本)9781510648043;9781510648036

The triple-negative breast cancer (tnbc) is an aggressive subtype linked to a poor outcome of established breast cancer therapy. Increasing evidence points to the role of the tumor's extracellular matrix (ECM) as a determinant of its aggressiveness as well as the effectiveness of chemical therapeutics. Three-dimensional imaging techniques can be used to unravel ECM architecture. Label-free contrast mechanisms such as second harmonic generation (SHG) avoid falsification and artifacts introduced by the labeling process. Here, we present the complementary use of two-photon excitation microscopy (TPEF) and Scanning Laser Optical Tomography (SLOT) for the investigation and quantification of tumor ECM. Both methods were used to capture fluorescence from antibody-labeled samples as well as the SHG signal from collagen strands in the ECM. SLOT generally allows for the investigation of larger samples of several mm up to a few cm in size. This work shows the capabilities of the tomographic setup compared to established TPEF, and demonstrates their combined use to maximize the information content of the acquired data. The obtained images served as a basis for ECM quantification. 3D-analysis allowed for determination of length, straightness and orientation of the collagen fibers based on fluorescence imaging as well as SHG imaging. The resulting coordinates might be used for synthetic reconstruction of a patient-specific tumor matrix, serving as a scaffold for pre-clinical therapeutic testing. Collagen imaging and quantification as presented here can therefore be employed for both basic and clinical research, paving the way for patient-specific cancer therapy.

关键词： cancer collagen imaging multimodal imaging extracellular matrix 3D imaging image reconstruction tomography second harmonic generation

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