near real-time deconvolution for 3d fluorescence microscopy...near real-time deconvolution for 3d...
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![Page 1: Near Real-Time Deconvolution for 3D Fluorescence Microscopy...Near Real-Time Deconvolution for 3D Fluorescence Microscopy Marc Bruce, PhD CEO, Microvolution ... faster than old methods](https://reader034.vdocuments.site/reader034/viewer/2022042806/5f6df957fb9b6927d80cfd6d/html5/thumbnails/1.jpg)
Near Real-Time Deconvolution for 3D Fluorescence Microscopy
Marc Bruce, PhD
CEO, Microvolution
![Page 2: Near Real-Time Deconvolution for 3D Fluorescence Microscopy...Near Real-Time Deconvolution for 3D Fluorescence Microscopy Marc Bruce, PhD CEO, Microvolution ... faster than old methods](https://reader034.vdocuments.site/reader034/viewer/2022042806/5f6df957fb9b6927d80cfd6d/html5/thumbnails/2.jpg)
GPU Accelerated Deconvolution Software
Microvolution offers faster and more accurate deconvolution
Thin Filaments Are Missing Preserves Thin Filaments: 16 sec
Image Courtesy of Molecular Devices
Other Deconvolution10.7 Min
After 3-D Deconvolution 16 Seconds
![Page 3: Near Real-Time Deconvolution for 3D Fluorescence Microscopy...Near Real-Time Deconvolution for 3D Fluorescence Microscopy Marc Bruce, PhD CEO, Microvolution ... faster than old methods](https://reader034.vdocuments.site/reader034/viewer/2022042806/5f6df957fb9b6927d80cfd6d/html5/thumbnails/3.jpg)
Basics of Fluorescence Microscopy
Sample
Objective
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Basics of Fluorescence Microscopy
IlluminationDichroic mirror
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Basics of Fluorescence Microscopy
Emission
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Basics of Fluorescence Microscopy
• Damage to sample from high intensity light
• Haze from out-of-plane fluorescence
• Blurriness due to diffraction reduces resolution of small objects
Derived from Wikipedia, “Light sheet fluorescence microscopy”
![Page 7: Near Real-Time Deconvolution for 3D Fluorescence Microscopy...Near Real-Time Deconvolution for 3D Fluorescence Microscopy Marc Bruce, PhD CEO, Microvolution ... faster than old methods](https://reader034.vdocuments.site/reader034/viewer/2022042806/5f6df957fb9b6927d80cfd6d/html5/thumbnails/7.jpg)
Diffraction Limits 3D Resolution
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Point Spread Function
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The Solution: Deconvolution Software
http://www.leica-microsystems.com/science-lab/deconvolution
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Richardson Lucy Algorithm
Iterative algorithm operating under the assumptions of Poisson-distributed noise
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Real-time Deconvolution is Now Possible
• GPU yields speeds up to 200X faster than old methods
• Allows for real-time deconvolution • Optimize image capture
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Higher Signal-to-Noise Ratio Yields Better Data
• Analysis of raw FRET images showed low contrast
• 4x effective increase in contrast post-deconvolution
• Expected cell features can now be seen
Miskolci, V., Hodgson, L. & Cox, D. Using Fluorescence Resonance Energy Transfer-Based Biosensors to Probe Rho GTPase Activation During Phagocytosis. Methods in Molecular Biology 1519, 125-43 (2017)
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Diffraction-Limited Resolution Limit
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Increased Resolution to 180 nm
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Tong Zhang and Puifai Santisakultarm,Salk Institute
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Microscopy in Dim Light
Before
2048 x 2048 x 51
After 3D Deconvolution
8.5 seconds
Image Courtesy of Technical Instruments
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Widefield Microscopy
Dennis Hughes, MD, PhD, Assoc Prof of Pediatrics, Jared Mortus, Lead Researcher, Laurence Cooper, MD, PhD, Professor of Pediatrics, George McNamara, PhD, Senior Research Scientist, The Children’s Cancer Hospital, MD Anderson Cancer Center, Houston, TX
Widefield Image Before
1024 x 1024 x 34
After 3D Deconvolution
1.6 seconds
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Light Sheet Microscopy
Derived from Wikipedia, “Light sheet fluorescence microscopy”
![Page 18: Near Real-Time Deconvolution for 3D Fluorescence Microscopy...Near Real-Time Deconvolution for 3D Fluorescence Microscopy Marc Bruce, PhD CEO, Microvolution ... faster than old methods](https://reader034.vdocuments.site/reader034/viewer/2022042806/5f6df957fb9b6927d80cfd6d/html5/thumbnails/18.jpg)
Light Sheet Microscopy
Light Sheet Image Before
946 x 768 x 321
After 3D Deconvolution
22.5 seconds
Dong-Yuan Chen in the Bilder lab, UC Berkeley
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Lattice Light Sheet Microscopy
Dr. Boyd Butler using 3i Lattice LightSheet
LLSM Before
1167 x 512 x 401
After 3D Deconvolution
10.2 seconds
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Instant SIM
VisiTech
Instant SIM Image Before
1446 x 1131 x 14
After 3D Deconvolution
4.9 seconds
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FINCH (Fresnel incoherent correlation holography)
Siegel, N., Lupashin, V., Storrie, B. & Brooker, G. High-magnification super-resolution FINCH microscopy using birefringent crystal lens interferometers. Nature Photonics 10, 802-808 (2016)
After Propagation and 3D Deconvolution
10 seconds
Widefield Image
2048 x 2048 x 21
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Usable on All Tiers of GPUs
• HPC clusters• Split large multi-channel, multi-timepoint images onto independent nodes
• Let peer GPUs collaborate
• Desktops• Run alongside acquisition
• Embedded systems• Currently developing for TX1
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Usable on All Tiers of GPUs
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