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

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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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Validation of Excitation-Scan Hyperspectral Multi-faceted Mirror Array Prototype System Advancements to Hyperspectral Imaging Applications 29

Validation of Excitation-Scan Hyperspectral Multi-faceted Mi...

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

作者： Parker, Marina Mayes, Sam A. Browning, Craig M. Deal, Joshua Gunn-Mayes, Samantha Rich, Thomas C. Leavesley, Silas J. Univ S Alabama Chem & Biomol Engn Mobile AL 36688 USA Univ S Alabama Pharmacol Mobile AL 36688 USA Univ S Alabama Ctr Lung Biol Mobile AL 36688 USA Univ S Alabama Dept Syst Engn Mobile AL 36688 USA

ISBN: (纸本)9781510648043;9781510648036

Hyperspectral imaging technologies (HSI) have undergone rapid development since their beginning stages. While original applications were in remote sensing, other uses include agriculture, food safety and medicine. HSI has shown great utility in fluorescence microscopy for detecting signatures from many fluorescent molecules;however, acquisitions speeds have been slow due to light losses associated with spectral filtering. Therefore, we designed a novel light emitting diode (LED)-based rapid excitation scanning hyperspectral imaging platform allowing users to obtain simultaneous measurements of fluorescent labels without compromising acquisition speeds. Previously, we reported our results of the optical ray trace simulations and the geometrical capability of designing a multifaceted mirror imaging system as an initial approach to combine light at many wavelengths. The design utilized LEDs and a multifaceted mirror array to combine light sources into a liquid light guide. The computational model was constructed using Monte Carlo optical ray software (TracePro, Lambda Research Corp.). Recent prototype validation results show that when compared to a commercial emission scanning spectral confocal microscope (Zeiss-LSM-980), the novel LED-based excitation scanning HSI prototype successfully detected and separated six fluorescent labels from a custom 6-label African green monkey kidney epithelial cells. We report on the prototype's ability to overcome limitations of acquisition speeds, sensitivity, and specificity present in conventional systems. Future work will evaluate prototype's light losses to determine latent design modifications needed to demonstrate the system's feasibility as a promising solution for overcoming HSI acquisition speeds. This work was supported by NSF award MRI1725937.

关键词： Systems Design LED Tissue imaging HSI Spectroscopy microscopy Transmission Spectral Fluorescence Molecules Cells

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A study on image processing Method for Noise Reduction in Digital Holographic microscopy 9

A study on Image Processing Method for Noise Reduction in Di...

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9th International conference on Intelligent Informatics and BioMedical Sciences, ICIIBMS 2024

作者： Nagashima, Masaharu Kim, Hyun-Woo Cho, Myungjin Lee, Min-Chul Kyushu Institute of Technology Graduate School of Computer Science and Systems Engineering Iizuka city Japan Iitc Hankyong National University School of Ict Robotics and Mechanical Engineering Anseong-si Korea Republic of

ISBN: (纸本)9798350363043

Digital Holographic microscopy (DHM) can obtain three-dimensional (3D) information about the fine structure of an object by utilizing the phase information of coherent light. In DHM, during the process of reconstructing the shape of an object, phase information is extracted by windowing the sideband containing the phase information. The 3D information of the object is then obtained from the acquired phase information. When windowing the sideband in a narrow range, it can remove noise caused by the DC spectrum, but detailed shape information of the object is lost. On the other hand, when windowing in a wide range, detailed shape information of the object can be obtained, but noise from the DC spectrum increases. To solve this trade-off relationship, we propose a method of determining the range for windowing the sideband using a function and then averaging the windowed sidebands. This proposed method can reduce the impact of noise from the DC spectrum and enables the acquisition of the detailed shape information. In this paper, we explain the reconstruction algorithm of this method and compare it with conventional methods and the previous study, phase differences averaging (PDA). © 2024 IEEE.

关键词： Noise abatement

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Hyperspectral imaging and adaptive thresholding to identify agonist-induced cAMP signals in pulmonary microvascular endothelial cells 29

Hyperspectral imaging and adaptive thresholding to identify ...

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

作者： Annamdevula, N. Johnson, S. Pleshinger, D. J. Castleberry, S. Russell, W. Britain, A. L. Francis, C. M. Rich, T. C. Leavesley, S. J. Univ S Alabama Pharmacol Mobile AL 36688 USA Univ S Alabama Physiol & Cell Biol Mobile AL 36688 USA Univ S Alabama Chem & Biomol Engn Mobile AL 36688 USA Univ West Alabama Livingston AL 35470 USA Auburn Univ Auburn AL 36849 USA

ISBN: (纸本)9781510648043;9781510648036

Cyclic AMP (cAMP) is a second messenger that regulates a wide variety of cellular functions. There is increasing evidence suggesting that signaling specificity is due in part to cAMP compartmentalization. In the last 15 years, development of cAMP-specific Forster resonance energy transfer (FRET) probes have allowed us to visualize spatial distributions of intracellular cAMP signals. The use of FRET-based sensors is not without its limitations, as FRET probes display low signal to noise ratio (SNR). Hyperspectral imaging and analysis approaches have, in part, allowed us to overcome these limitations by improving the SNR of FRET measurements. Here we demonstrate that the combination of hyperspectral imaging approaches, linear unmixing, and adaptive thresholding allow us to visualize regions of elevated cAMP (regions of interest - ROIs) in an unbiased manner. We transfected cDNA encoding the H188 FRET-based cAMP probe into pulmonary microvascular endothelial cells. Application of isoproterenol and prostaglandin E1 (PGE(1)) triggered complex cAMP responses. Spatial and temporal aspects of cAMP responses were quantified using an adaptive thresholding approach and compared between agonist treatment groups. Our data indicate that both the origination sites and spatial/temporal distributions of cAMP signals are agonist dependent in PMVECs. We are currently analyzing the data in order to better quantify the distribution of cAMP signals triggered by different agonists.

关键词： FRET fluorescence microscopy spectral imaging adaptive thresholding second messengers cAMP signals localized signals

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Multi-immersion open-top light-sheet microscopy 27

Multi-immersion open-top light-sheet microscopy

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

作者： Glaser, Adam K. Liu, Jonathan T. C. Univ Washington Dept Mech Engn Seattle WA 98195 USA

We present an easy-to-use multi-immersion open-top light-sheet microscope designed specifically for high-throughput imaging of a diverse set of tissues prepared with a variety of clearing protocols.

ISBN: (纸本)9781510632547

We present an easy-to-use multi-immersion open-top light-sheet microscope designed specifically for high-throughput imaging of a diverse set of tissues prepared with a variety of clearing protocols.

关键词： light-sheet microscopy selective-plane illumination microscopy fluorescence microscopy

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A study for accelerating the speed of all-in-focus image processing 27

A study for accelerating the speed of all-in-focus image pro...

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

作者： Wang, Lihui Tabata, Satoshi Takeuchi, Hirotoshi Ishikawa, Masatoshi Guangdong Acad Sci Guangdong Inst Semicond Ind Technol 363 Changxing Rd Guangzhou 510650 Peoples R China Univ Tokyo Grad Sch Informat Sci & Technol Bunkyo Ku 7-3-1 Hongo Tokyo 1138656 Japan

ISBN: (纸本)9781510632547

To capture an all-in-focus and 3D depth image, shape from focus method is widely used. The phase accuracy of the image candidates should be processed and adjusted beforehand Phase only correction method was employed in this work and to accelerate the processing speed, the image processing by Fast Fourier Transform (FFT) was optimized Meanwhile, the processing task was assigned in several parallel threads so that the performance would be improved. The method was used on a variable focus imaging system, and, as a result, the processing speed was improved to around 2.5-fps.

关键词： all-in-focus shape from focus phase-only correction method 3D imaging variable focus system

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3,3′-thiodipropanol as a refractive index matching mounting medium for fluorescence microscopy 27

3,3′-thiodipropanol as a refractive index matching mounting...

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

作者： Alata Tejedo, Milvia Iris Martinez Cervantes, Juan Carlos Piazza, Valeria Ctr Invest Opt AC Loma Bosque 115 Guanajuato Mexico

ISBN: (纸本)9781510632547

In this work, several tests of the performance of 3,3'-thiodipropanol (TDP) as a mounting medium for fluorescence microscopy of biological samples were performed. Besides optical properties like the dispersion curve of TDP and the effect of the embedding medium on the fluorescence of commonly used dyes, the interaction with biological specimens, including the labeling of filamentous actin with fluorescent phalloidins, was tested: TDP showed to represent an interesting alternative to commercial mounting media. By mixing TDP with 2,2'-thiodiethanol (TDE), it was possible not only to fine tune the refractive index of the resulting solution, but to preserve the compatibility with fluorescent phalloidins.

关键词： Mounting media fluorescence microscopy filamentous actin

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Computational super-resolution microscopy: leveraging noise model, regularization and sparsity to achieve highest resolution 27

Computational super-resolution microscopy: leveraging noise ...

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

作者： Xing, Jian Chen, Simeng Becker, Stephen Yu, Jiun-Yann Cogswell, Carol Univ Colorado Dept Elect Comp & Energy Engn Boulder CO 80309 USA Univ Colorado Dept Appl Math Boulder CO 80309 USA

ISBN: (纸本)9781510632547

We report progress in algorithm development for a computation-based super-resolution microscopy technique. Building upon previous results, we examine our recently implemented microscope system and construct alternative processing algorithms. Based on numerical simulations results, we evaluate the performance of each algorithm and determine the one most suitable for our super-resolution microscope.

关键词： Super-resolution microscopy regularization computational microscopy

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Multifocus imaging via polarization wavefront shaping 27

Multifocus imaging via polarization wavefront shaping

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

作者： Li, Chen Ding, Changqin Li, Minghe Rong, Jiayue Ulcickas, James Simpson, Garth J. Purdue Univ Dept Chem W Lafayette IN 47907 USA

ISBN: (纸本)9781510632547

Conventional three-dimensional (3D) images of biological samples are typically assembled from a stack of two-dimensional images acquired sequentially at different focal planes. This time-consuming manner hinders the application of 3D imaging techniques to the investigation of fast biochemical dynamics and light-sensitive biological events. The concept of multifocus imaging, which enables simultaneous acquisition of images from multiple focal planes, was introduced to achieve rapid 3D imaging. In the present study, we achieved multifocus imaging through polarization wavefront shaping via a micro-retarder array which splits the incident linearly polarized light into three beamlets that are focused to three axially-offset focal planes with 100 itm separation. Append to an existing beam-scanning microscope, this multifocus system enables rapid 3D imaging compatible with a variety of optical microscopic approaches including laser transmittance, two-photon excited fluorescence, and second harmonic generation microscopy.

关键词： Multifocus imaging beam scanning microscopy polarization wavefront shaping Jones calculus

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PHASE RETRIEVAL FROM 4-dimensional ELECTRON DIFFRACTION DATASETS

PHASE RETRIEVAL FROM 4-DIMENSIONAL ELECTRON DIFFRACTION DATA...

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IEEE International conference on image processing (ICIP)

作者： Friedrich, Thomas Yu, Chu-Ping Verbeek, Johan Pennycook, Timothy Van Aert, Sandra Univ Antwerp Electron Microscopy Mat Sci EMAT Groenenborgerlaan 171 B-2020 Antwerp Belgium Univ Antwerp NANOlab Ctr Excellence Groenenborgerlaan 171 B-2020 Antwerp Belgium

ISBN: (纸本)9781665441155

We present a computational imaging mode for large scale electron microscopy data, which retrieves a complex wave from noisy/sparse intensity recordings using a deep learning approach and subsequently reconstructs an image of the specimen from the Convolutional Neural Network (CNN) predicted exit waves. We demonstrate that an appropriate forward model in combination with open data frameworks can be used to generate large synthetic datasets for training. In combination with augmenting the data with Poisson noise corresponding to varying dose-values, we effectively eliminate overfitting issues. The U-NET[1] based architecture of the CNN is adapted to the task at hand and performs well while maintaining a relatively small size and fast performance. The validity of the approach is confirmed by comparing the reconstruction to well-established methods using simulated, as well as real electron microscopy data. The proposed method is shown to be effective particularly in the low dose range, evident by strong suppression of noise, good spatial resolution, and sensitivity to different atom types, enabling the simultaneous visualisation of light and heavy elements and making different atomic species distinguishable. Since the method acts on a very local scale and is comparatively fast it bears the potential to be used for near-real-time reconstruction during data acquisition.

关键词： phase retrieval inverse problem electron diffraction 4D-STEM CBED

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