scholarly journals HyU: Hybrid Unmixing for longitudinal in vivo imaging of low signal to noise fluorescence

2022 ◽  
Author(s):  
Hsiao Chiang ◽  
Daniel Koo ◽  
Masahiro Kitano ◽  
Jay Unruh ◽  
Le Trinh ◽  
...  
Keyword(s):  
Dynamic Range ◽  
Cell Metabolism ◽  
High Dynamic Range ◽  
Dynamic Imaging ◽  
Biological Research ◽  
Fluorescent Labels ◽  
Volumetric Imaging ◽  
Linear Unmixing ◽  
Cellular Behaviors

Abstract The expanded application of fluorescence imaging in biomedical and biological research towards more complex systems and geometries requires tools that can analyze a multitude of components at widely varying time- and length-scales. The major challenge in such complex imaging experiments is to cleanly separate multiple fluorescent labels with overlapping spectra from one another and background autofluorescence, without perturbing the sample with high levels of light. Thus, there is a requirement for efficient and robust analysis tools capable of quantitatively separating these signals. In response, we have combined multispectral fluorescence microscopy with hyperspectral phasors and linear unmixing to create Hybrid Unmixing (HyU). Here we demonstrate its capabilities in the dynamic imaging of multiple fluorescent labels in live, developing zebrafish embryos. HyU is more sensitive to low light levels of fluorescence compared to conventional linear unmixing approaches, permitting better multiplexed volumetric imaging over time, with less bleaching. HyU can also simultaneously image both bright exogenous and dim endogenous labels because of its high dynamic range. This allows studies of cellular behaviors, tagged components, and cell metabolism within the same specimen, offering a powerful window into the orchestrated complexity of biological systems.

scholarly journals Methods for Voltage-Sensitive Dye Imaging of Rat Cortical Activity With High Signal-to-Noise Ratio

2007 ◽  
Vol 98 (1) ◽  
pp. 502-512 ◽  
Author(s):  
Michael T. Lippert ◽  
Kentaroh Takagaki ◽  
Weifeng Xu ◽  
Xiaoying Huang ◽  
Jian-Young Wu
Keyword(s):  
Dynamic Range ◽  
Signal To Noise Ratio ◽  
Field Potential ◽  
High Sensitivity ◽  
High Dynamic Range ◽  
High Signal ◽  
Blue Dye ◽  
Voltage Sensitive Dye ◽  
Imaging Device

We describe methods to achieve high sensitivity in voltage-sensitive dye (VSD) imaging from rat barrel and visual cortices in vivo with the use of a blue dye RH1691 and a high dynamic range imaging device (photodiode array). With an improved staining protocol and an off-line procedure to remove pulsation artifact, the sensitivity of VSD recording is comparable with that of local field potential recording from the same location. With this sensitivity, one can record from ∼500 individual detectors, each covering an area of cortical tissue 160 μm in diameter (total imaging field ∼4 mm in diameter) and a temporal resolution of 1,600 frames/s, without multiple-trial averaging. We can record 80–100 trials of intermittent 10-s trials from each imaging field before the VSD signal reduces to one half of its initial amplitude because of bleaching and wash-out. Taken together, the methods described in this report provide a useful tool for visualizing evoked and spontaneous waves from rodent cortex.

scholarly journals In-vivo high dynamic range vector flow imaging

2015 ◽  
Author(s):  
Carlos A. Villagomez-Hoyos ◽  
Matthias Bo Stuart ◽  
Jorgen Arendt Jensen
Keyword(s):  
Dynamic Range ◽  
High Dynamic Range ◽  
Flow Imaging ◽  
Range Vector ◽  
High Dynamic

scholarly journals Attention Mechanism Based Semi-Supervised Multi-Gain Image Fusion

Symmetry ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 451
Author(s):  
Ming Fang ◽  
Xu Liang ◽  
Feiran Fu ◽  
Yansong Song ◽  
Zhen Shao
Keyword(s):  
Image Fusion ◽  
Network Structure ◽  
Dynamic Range ◽  
High Dynamic Range ◽  
Dynamic Imaging ◽  
Motion Artifacts ◽  
Attention Mechanism ◽  
Source Image ◽  
Fused Image ◽  
High Dynamic

High-dynamic range imaging technology is an effective method to improve the limitations of a camera’s dynamic range. However, most current high-dynamic imaging technologies are based on image fusion of multiple frames with different exposure levels. Such methods are prone to various phenomena, for example motion artifacts, detail loss and edge effects. In this paper, we combine a dual-channel camera that can output two different gain images simultaneously, a semi-supervised network structure based on an attention mechanism to fuse multiple gain images is proposed. The proposed network structure comprises encoding, fusion and decoding modules. First, the U-Net structure is employed in the encoding module to extract important detailed information in the source image to the maximum extent. Simultaneously, the SENet attention mechanism is employed in the encoding module to assign different weights to different feature channels and emphasis important features. Then, a feature map extracted from the encoding module is input to the decoding module for reconstruction after fusing by the fusion module to obtain a fused image. Experimental results indicate that the fused images obtained by the proposed method demonstrate clear details and high contrast. Compared with other methods, the proposed method improves fused image quality relative to several indicators.

scholarly journals Optical Control of Cytokine Signaling via Bioinspired, Polymer-Induced Latency

2020 ◽  
Author(s):  
Lacey A Perdue ◽  
Priscilla Do ◽  
Camille David ◽  
Andrew Chyong ◽  
Anna Kellner ◽  
...  
Keyword(s):  
Dynamic Range ◽  
High Dynamic Range ◽  
Optical Control ◽  
Signaling Networks ◽  
Cytokine Signaling ◽  
Receptor Subunit ◽  
Protein Activity ◽  
Signal Specificity ◽  
High Dynamic

ABSTRACTCytokine signaling is challenging to study and therapeutically exploit as the effects of these protein are often pleiotropic. A subset of cytokines can, however, exert signal specificity via association with latency-inducing proteins which cage the cytokine until disrupted by discreet biological stimuli. Inspired by this precision, here we describe a strategy for synthetic induction of cytokine latency via modification with photo-labile polymers that mimic latency while attached, then restore protein activity in response to light, thus controlling the magnitude, duration, and location of cytokine signals. We characterize the high dynamic range of latent cytokine activity modulation and find that polymer-induced latency, alone, can prolong in vivo circulation and bias receptor subunit binding. We further show that protein de-repression can be achieved with near single-cell resolution and demonstrate the feasibility of transcutaneous photoactivation. Future extensions of this approach could enable multicolor, optical reprogramming of cytokine signaling networks and more precise immunotherapies.

Description and visualization of the highly dynamic behavior of the electrorheological effect

2019 ◽  
Vol 31 (2) ◽  
pp. 308-317
Author(s):  
Tobias Bauerochs ◽  
Xiaoye Huo ◽  
Gilad Yossifon ◽  
Stephan Ulrich ◽  
Steffen Schneider ◽  
...  
Keyword(s):  
Electric Field ◽  
High Speed ◽  
Dynamic Range ◽  
Electrorheological Fluid ◽  
High Dynamic Range ◽  
Dynamic Imaging ◽  
Particle Behavior ◽  
Electrorheological Effect ◽  
High Dynamic ◽  
Field Lines

When an electrorheological fluid is located between two electrodes and an electrical voltage is applied to them, the particles in the fluid move and form chains along the electric field lines. This phenomenon is called the electrorheological effect. The exact behavior of the particles has not yet been studied completely. Some optical investigations of particle motion or behavior have been performed, but did not take into account the high dynamic range directly after the application of an electric field. This study is intended to help explain how the particles behave when they encounter an electric field and then try to align themselves with it. There is an investigation into how these chains develop in a microchannel within milliseconds. For this purpose, the particle behavior of the electrorheological fluid is investigated with high dynamic imaging using a microscope. A high-speed camera records videos of the first milliseconds at 3000 fps synchronously with the application of an electric field. The results provide a better understanding of the chain formation and particle behavior of the electrorheological effect in the high dynamic range and can be used for the design of electrorheological applications as well as simulations of the particle movement.

scholarly journals In vivo imaging of individual islets across the mouse pancreas reveals a heterogeneous insulin secretion response to glucose

2020 ◽  
Author(s):  
Henriette Frikke-Schmidt ◽  
Peter Arvan ◽  
Randy J Seeley ◽  
Corentin Cras-Méneur
Keyword(s):  
Insulin Secretion ◽  
Dynamic Range ◽  
High Dynamic Range ◽  
Motion Blur ◽  
Isolated Islets ◽  
Computer Assisted ◽  
Powerful Approach ◽  
Glucose Challenge ◽  
The Impact

AbstractWhile numerous techniques can be used to measure and analyze insulin secretion in isolated islets in culture, assessments of insulin secretion in vivo are typically indirect and only semiquantitative. The CpepSfGFP reporter mouse line allows the in vivo imaging of insulin secretion from individual islets after a glucose challenge, in live, anesthetized mice, addressing secretion from the pancreas as a whole. Imaging the whole pancreas at high resolution in live mice includes numerous technical challenges. Using rapid-fire imaging in high dynamic range and tiling combined with computer-assisted masked morphometry, we have developed a method to overcome motion blur, and the effects of anesthesia, to be able to monitor and quantify insulin (CpepSfGFP) content simultaneously and longitudinally for the first time in hundreds of individual islets, throughout a glucose challenge.Through this approach we demonstrate that while isolated islets respond homogeneously to glucose in culture, their response profile differs significantly in vivo. Independent of size or location, some islets respond sharply to a glucose stimulation while others barely secrete at all. This platform therefore provides a powerful approach to study the impact of disease, diet, surgery or pharmacological treatments on insulin secretion in the intact pancreas in vivo.

scholarly journals Combined Atomic Force Microscope and Volumetric Light Sheet System for Mechanobiology

2019 ◽  
Author(s):  
E. Nelsen ◽  
C.M. Hobson ◽  
M.E. Kern ◽  
J.P. Hsiao ◽  
E.T. O’Brien ◽  
...  
Keyword(s):  
Atomic Force Microscope ◽  
Dynamic Range ◽  
Light Sheet ◽  
High Dynamic Range ◽  
Actin Dynamics ◽  
Mechanical Stimuli ◽  
Spatiotemporal Resolution ◽  
Volumetric Imaging ◽  
Novel Studies ◽  
Atomic Force

ABSTRACTThe central goals of mechanobiology are to understand how cells generate force and how they respond to environmental mechanical stimuli. A full picture of these processes requires high-resolution, volumetric imaging with time-correlated force measurements. Here we present an instrument that combines an open-top, single-objective light sheet fluorescence microscope with an atomic force microscope (AFM), providing simultaneous volumetric imaging with high spatiotemporal resolution and high dynamic range force capability (10 pN – 100 nN). With this system we have captured lysosome trafficking, vimentin nuclear caging, and actin dynamics on the order of one second per volume. To showcase the unique advantages of combining Line Bessel light sheet imaging with AFM, we measured the forces exerted by a macrophage during FcɣR-mediated phagocytosis while performing both sequential two-color, fixed plane and volumetric imaging of F-actin. This unique instrument allows for a myriad of novel studies investigating the coupling of cellular dynamics and mechanical forces.

High-dynamic-range interferometric bragg-cell spectrum analyser

1986 ◽  
Vol 133 (1) ◽  
pp. 26
Author(s):  
J. Mellis ◽  
G.R. Adams ◽  
K.D. Ward
Keyword(s):  
Dynamic Range ◽  
High Dynamic Range ◽  
Spectrum Analyser ◽  
High Dynamic
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