Orca flash 4.0 cmos

Manufactured by Hamamatsu Photonics

The ORCA Flash 4.0 CMOS is a high-performance scientific CMOS image sensor designed for a variety of imaging applications. It features a large active area, high-speed readout, and low noise characteristics.

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Lab products found in correlation

Axio observer z1, by Zeiss (3 mentions) Orca r2 ccd, by Hamamatsu Photonics (1 mentions) Perfectfocus system, by Sutter Instruments (1 mentions) Plan fluor 20x, by Nikon (1 mentions) Eclipse ti inverted microscope, by Nikon (1 mentions) Metamorph software, by Molecular Devices (1 mentions) Plan apo ph na 1, by Yokogawa (1 mentions)

4 protocols using orca flash 4.0 cmos

Imaging Techniques for Hermaphrodite Worms

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Adult hermaphrodites were anesthetized with tricaine/tetramisole in PBS and immobilized between a coverslip and agarose pad on a slide. Imaging for Figs. 1, 2, 3, 4A, and 5G was done using MicroManager with a microscope (IX81; Olympus) equipped with an oil objective (60× PlanApo 1.42), a disk-scanning unit (Olympus), and a camera (ORCA Flash 4.0 CMOS; Hamamatsu Photonics). Imaging for Figs. 4 (C and D) and 5 (A–F, J, and K) were captured using Velocity on a Cetus Ultraview Spinning Disk Confocal microscope (PerkinElmer) equipped with a camera (Orca R2 CCD; Hamamatsu Photonics) and an oil objective (100× 1.35; Olympus). Fig. 2 (A–C) was deconvolved using Huygens Professional X11 (SVI). Fig. 5 G, Fig. S1, and Videos 1–5 were processed using ImageJ (National Institutes of Health) Rolling Ball Background Subtraction.

Panzica M.T., Marin H.C., Reymann A.C, & McNally F.J. (2017). F-actin prevents interaction between sperm DNA and the oocyte meiotic spindle in C. elegans. The Journal of Cell Biology, 216(8), 2273-2282.

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Intravital Microscopy of Microvascular Flow

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Microscopy studies used a Nikon TiE inverted epifluorescence microscope equipped with Nikon Perfect Focus System, Xenon arc lamp, Sutter Lambda 10-3 filter controller and Hamamatsu Orca Flash 4.0 CMOS detector. Images were collected at 97.5 fps, unbinned, at 0.65 μm/pixel (Figures 2,3) or binned 2×2 to 1.3 μm/pixel (Figure 4). All studies used a Nikon Plan Fluor 20X, NA 0.75 multi-immersion objective, water immersion mode. Since the STAFF macro measures microvascular flow in a fixed set of regions, organ stability is crucial. Organ immobilization methods for intravital microscopy are described in our previous publications^{2 (link),4 (link)}. Field of view may shift briefly during respiration, but returns to its original position after each respiration, allowing reliable measurements.

Clendenon S.G., Fu X., Von Hoene R.A., Clendenon J.L., Sluka J.P., Winfree S., Mang H., Martinez M., Filson A.J., Klaunig J.E., Glazier J.A, & Dunn K.W. (2018). A simple automated method for continuous fieldwise measurement of microvascular hemodynamics. Microvascular research, 123, 7-13.

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Fluorescence Imaging Microscopy Protocol

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All immunostaining and live-imaging experiments were performed on a Nikon Eclipse Ti–inverted microscope and imaged using a 10×/0.30 NA plan Fluor, 40×/1.30 NA plan Fluor, 100×/1.40 NA plan APO, or 100×/1.49 NA TIRF objective, CSU-22 spinning disc confocal module (Yokogawa), and either an ORCA-Flash4.0 CMOS (Hamamatsu) or interline transfer-cooled charge coupled device camera (CoolSNAP HQ2; Photometrics). 491-, 561-, and 642-nm solid-state lasers were used for excitation (VisiTech International), and a MAC6000 Automation Controller (Ludl Electronic Products) was used to operate an emission filter wheel equipped with Semrock Emission Filters. Metamorph software (v7.7.10; Molecular Devices) was used for image acquisition.

Schoborg T., Zajac A.L., Fagerstrom C.J., Guillen R.X, & Rusan N.M. (2015). An Asp–CaM complex is required for centrosome–pole cohesion and centrosome inheritance in neural stem cells. The Journal of Cell Biology, 211(5), 987-998.

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High-Resolution 3D Confocal Imaging of Cells

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To obtain high resolution three-dimensional (3D) images of the cells, a z-stack of confocal images was acquired using a spinning disk confocal imaging system based on a Zeiss Axio Observer Z1 inverted fluorescence microscope (with 63x Plan Apo PH NA 1.4), equipped with a computer-controlled Spherical Aberration Correction unit, Yokogawa CSU-X1, Vector photomanipulation module, Photometrics Evolve 16-bit EMCCD and Hamamatsu Orca-Flash4.0 CMOS cameras, environmental chamber and piezo stage controller and lasers (405, 445, 488, 515, 561, and 640 nm), all controlled by SlideBook six software (Intelligent Imaging Innovation, Denver, CO). Typically, 15–30 serial two-dimensional confocal images were recorded at 200–400 nm intervals. All image acquisition settings were identical for all experimental variants in each experiment.

Sorkina T., Ma S., Larsen M.B., Watkins S.C, & Sorkin A. (2018). Small molecule induced oligomerization, clustering and clathrin-independent endocytosis of the dopamine transporter. eLife, 7, e32293.

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