Csu 10

Manufactured by Zeiss

The Zeiss CSU-10 is a compact and versatile confocal scanning unit designed for high-speed, high-resolution imaging. It utilizes a spinning disk to provide rapid optical sectioning, enabling the capture of live-cell dynamics with minimal phototoxicity. The CSU-10 can be readily integrated with a wide range of microscopes, making it a valuable tool for a variety of applications in the life sciences.

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Lab products found in correlation

Orca em ccd camera, by Zeiss (2 mentions) Metamorph software, by Molecular Devices (2 mentions) Axio imager a2, by Zeiss (2 mentions) Axioimager microscope, by Zeiss (1 mentions) Imagej 1.40g, by Adobe (1 mentions) Em ccd camera, by Yokogawa (1 mentions) Plan apochromat objective, by Zeiss (1 mentions) Celltram vario, by Eppendorf (1 mentions) Em ccd camera, by Hamamatsu Photonics (1 mentions) Dmi6000b inverted microscope, by Yokogawa (1 mentions) Axioimager, by Hamamatsu Photonics (1 mentions) Puromycin, by Santa Cruz Biotechnology (1 mentions) Halocarbon oil 27, by Merck Group (1 mentions) α amanitin, by Santa Cruz Biotechnology (1 mentions) Halocarbon oil 700, by Merck Group (1 mentions) Mn 151, by Narishige (1 mentions) Plan apochromat 100x 1.4 oil dic objective, by Zeiss (1 mentions) Plan apochromat 100x 1.4 na oil dic objective, by Zeiss (1 mentions) Axiocam mrm camera, by Zeiss (1 mentions)

5 protocols using csu 10

Confocal Microscopy Imaging and Analysis

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Images were acquired using a Hamamatsu EM-CCD camera and a Yokogawa CSU-10 spinning disk confocal mounted on a Zeiss AxioImager microscope with a 100× Plan-Apochromat objective. The microscope was controlled by iVision software (Biovision Technologies, Exton, PA) or microManager (Edelstein et al., 2010 ). Acquired images were processed to enhance brightness/contrast using ImageJ 1.40g and Photoshop (CS6 Extended Adobe Systems, InC., San Jose, CA), and smoothened using a 0.8 pixel radius Gaussian blur filter. 3D reconstructions were built from confocal Z-stacks, analyzed, and exported as (.mov) files using IMARIS 7.4 (Bitplane, Inc., Saint Paul, MN). Figures and graphs were constructed using Illustrator (CS3 Extended Adobe Systems, Inc., San Jose, CA) and JMP (Version 10, SAS Institute Inc., Cary, NC, 1989-2007). Movies were annotated using Photoshop. Timelapse imaging and invadopodia analysis was performed as described previously (Hagedorn et al., 2013 (link)).

Morrissey M.A., Keeley D.P., Hagedorn E.J., McClatchey S.T., Chi Q., Hall D.H, & Sherwood D.R. (2014). B-LINK: A hemicentin, plakin and integrin-dependent adhesion system that links tissues by connecting adjacent basement membranes. Developmental cell, 31(3), 319-331.

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Zebrafish Xenotransplantation of STAT3 Variants

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All zebrafish experiments were approved by the Stony Brook University Institutional Animal Use and Care Committee. Two-day-old kdrl:RFP transgenic embryos were anesthetized with 0.016% tricaine and transferred to embryo medium containing 4% Ficoll. Fifty to 100 cells (control, STAT3 knockout, STAT3 Y640F, and STAT3 S727E) were injected into the common cardinal vein of the embryos using a CellTram Vario (Eppendorf) and transferred to a 33°C incubator. Embryos were imaged every 2 d after injection using a Leica DMI6000B inverted microscope and, on the eighth day after injection, with a spinning disc confocal microscope (Yokogawa, CSU-10; Carl Zeiss,AxioImager; Hamamatsu Photonics, EM-CCD camera).

D'Amico S., Shi J., Martin B.L., Crawford H.C., Petrenko O, & Reich N.C. (2018). STAT3 is a master regulator of epithelial identity and KRAS-driven tumorigenesis. Genes & Development, 32(17-18), 1175-1187.

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Live Imaging of Embryos with Pharmacological Manipulations

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Live imaging of embryos was conducted using a Yokogawa CSU10 spinning disc confocal microscope (Zeiss Observer.A1). Embryos were dechorionated in 4% bleach, washed with water, and mounted in halocarbon 27 Oil (Sigma). All live images were acquired in one minute intervals using Planapochrom 63X 1.4NA Oil objective. For pharmacological manipulations, embryos were lined up and glued to a coverslip, and desiccated for 12 to 13 minutes in a closed chamber containing drierite. Following desiccation, embryos were covered in halocarbon 700 oil (Sigma). Embryos were subsequently injected using a micromanipulator (Narishige, MN-151) with either water, 10,000 μg/mL puromycin (Santa Cruz Biotechnology), 1,000 μg/mL microbially sourced cycloheximide (Sigma), or 100 μg/mL α-amanitin (Santa Cruz Biotechnology) and imaged.

Patel P.H., Barbee S.A, & Blankenship J.T. (2016). GW-Bodies and P-Bodies Constitute Two Separate Pools of Sequestered Non-Translating RNAs. PLoS ONE, 11(3), e0150291.

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Automated Spinning Disk Confocal Imaging

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Images were collected using a custom-built spinning disk confocal microscope (Nobska Imaging), which was configured for automation with Metamorph software (Molecular Devices). This confocal consists of a Hamamatsu ORCA EM-CCD camera mounted on an upright Zeiss Axio Imager.A2 with a Borealis-modified Yokogawa CSU-10 spinning disk scanning unit and a Zeiss Plan-Apochromat 100x/1.4 oil DIC objective. Animals were anesthetized for imaging by picking them into a drop of M9 on a 5% agarose pad containing 7 mM sodium azide and secured with a coverslip.

Martinez M.A., Mullarkey A.A., Yee C., Zhao C.Z., Zhang W., Shen K, & Matus D.Q. (2022). Reevaluating the relationship between EGL-43 (EVI1) and LIN-12 (Notch) during C. elegans anchor cell invasion. Biology Open, 11(12), bio059668.

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Spinning Disk Confocal Imaging of C. elegans

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All micrographs included in this manuscript were collected on a Hamamatsu Orca EM-CCD camera mounted on an upright Zeiss AxioImager A2 with a Borealis-modified CSU10 Yokagawa spinning disk scan head using 405nm, 488 nm, and 561 nm Vortran lasers in a VersaLase merge and a Plan-Apochromat 100x/1.4 (NA) Oil DIC objective. MetaMorph software (Molecular Devices) was used for microscopy automation. Several experiments and all RNAi screening were scored using epifluorescence visualized on a Zeiss Axiocam MRM camera, also mounted on an upright Zeiss AxioImager A2 and a Plan-Apochromat 100x/1.4 (NA) Oil DIC objective. Animals were mounted into a drop of M9 on a 5% Noble agar pad containing approximately 10 mM sodium azide anesthetic and topped with a coverslip.

Smith J.J., Xiao Y., Parsan N., Medwig-Kinney T.N., Martinez M.A., Moore F.E., Palmisano N.J., Kohrman A.Q., Chandhok Delos Reyes M., Adikes R.C., Liu S., Bracht S.A., Zhang W., Wen K., Kratsios P, & Matus D.Q. (2022). The SWI/SNF chromatin remodeling assemblies BAF and PBAF differentially regulate cell cycle exit and cellular invasion in vivo. PLoS Genetics, 18(1), e1009981.

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