Csu w1 spinning disk

Manufactured by Yokogawa

Sourced in Japan, United States

The CSU-W1 spinning disk is a high-speed confocal scanning unit designed for live-cell imaging applications. It features a Nipkow-type spinning disk that rapidly scans the sample, enabling rapid image acquisition with low phototoxicity. The device is compatible with a wide range of microscopes and can be integrated into various imaging systems.

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Lab products found in correlation

Borealis system, by Oxford Instruments (6 mentions) Andor ixon ultra, by Oxford Instruments (5 mentions) Eclipse fn1 microscope, by Nikon (5 mentions) Metamorph software, by Molecular Devices (4 mentions) Ti e eclipse stand, by Nikon (2 mentions) Andor888 em ccd camera, by Oxford Instruments (2 mentions) 100 na 1.49 objective, by Nikon (2 mentions) Eclipse e600fn microscope, by Nikon (2 mentions) Cfi fluor lens, by Nikon (2 mentions) 1.0 cfi fluor lens, by Nikon (2 mentions) Inverted x83 microscope, by Hamamatsu Photonics (1 mentions) Prime 95b, by Yokogawa (1 mentions) Deltavision core system, by Cytiva (1 mentions) Eclipse ti microscope, by Nikon (1 mentions) Zyla 4.2 scmos camera, by Oxford Instruments (1 mentions) N sim e, by Nikon (1 mentions) Eclipse ti2 microscope, by Nikon (1 mentions) Lu n3 sim 488 561 640 laser unit, by Nikon (1 mentions) Orca flash 4.0 lt scmos camera, by Hamamatsu Photonics (1 mentions) Stage top incubation chamber, by Okolab (1 mentions)

24 protocols using csu w1 spinning disk

Live Tracking of HIV Integration in THP-1 Cells

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For all time-lapse studies, the cells were plated in a polymer-coverslip bottom µ-Dish 35 mm (ibidi #81156). THP-1 cells (2×10⁶) were differentiated for 48 h and then transduced with CPSF6 mNeonGreen LV (MOI = 0.01) for 3 days. Next, the cells were infected with HIV-1 and the live imaging was performed during 24–72 h p.i. For CPSF6 cluster fusion studies, CPSF6 mNeonGreen-positive cells were imaged every 5 min in 2D with a Biostation IM-Q (Nikon).
For FRAP experiments, the selected ROI was irradiated for 200 µs/pixel with 488 nm/561 nm laser and, after bleaching, the frames were acquired every 5–10 sec for 5 min through a Ti2E inverted microscope (Nikon), based on a CSU-W1 spinning-disk (Yokogawa), using a 60× objective (Plan Apochromat, oil immersion, NA = 1.4).
Experiments of live tracking of GIR and vDNA (OR-GFP) were performed in differentiated THP-1 cells transduced with OR-GFP LV (MOI = 5). Two days post-transduction, the cells were infected with HIV-1 ANCH3 GIR virus (MOI = 30). Different cells were imaged during 45–96 h p.i., every 5 min in 3D (stacks spacing 0.3 µm) with a Ti2E inverted microscope (Nikon), based on a CSU-W1 spinning-disk (Yokogawa), using a 60× objective (Plan Apochromat, oil immersion, NA = 1.4).

Scoca V., Morin R., Collard M., Tinevez J.Y, & Di Nunzio F. (2022). HIV-induced membraneless organelles orchestrate post-nuclear entry steps. Journal of Molecular Cell Biology, 14(11), mjac060.

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Measuring Cellular H2O2 Levels with HyPer

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HyPer (H₂O₂ ratiometric probe) fluorescence was excited with DPSS 10 mW 488 nm and 10 mW 405 nm lasers, and the corresponding YFP emission was measured using a 525/50 bandpass emission filter. Spinning-disk images were acquired using a 63× objective (63×/1.4 oil WD: 0.17 mm) on a Spinning-Disk CSU-W1 (Yokogawa) equipped with a sCMOS Hamamatsu 2048 × 2048 camera. To calculate the HyPer ratio, images were treated as previously described [57 (link)].

Thauvin M., Amblard I., Rampon C., Mourton A., Queguiner I., Li C., Gautier A., Joliot A., Volovitch M, & Vriz S. (2022). Reciprocal Regulation of Shh Trafficking and H2O2 Levels via a Noncanonical BOC-Rac1 Pathway. Antioxidants, 11(4), 718.

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Time-Lapse and Live Fluorescence Imaging

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Time-lapse bright-field imaging is done using an Olympus inverted X83 microscope with a Hamamatsu camera or a Spinning Disk CSU-W1 (Yokogawa) with a Prime 95B sCMOC camera. The latter is also used for live fluorescence imaging.

Pagès D.L., Dornier E., de Seze J., Gontran E., Maitra A., Maciejewski A., Wang L., Luan R., Cartry J., Canet-Jourdan C., Raingeaud J., Lemahieu G., Lebel M., Ducreux M., Gelli M., Scoazec J.Y., Coppey M., Voituriez R., Piel M, & Jaulin F. (2022). Cell clusters adopt a collective amoeboid mode of migration in confined nonadhesive environments. Science Advances, 8(39), eabp8416.

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Immunofluorescence Imaging of Collagen-I Cultured Cells

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TSIP#1 in Fig. 1B was fixed in 4% paraformaldehyde (PFA) after 24 hours of incubation in collagen I. Cell clusters in fig. S1F were fixed in PFA 4% for 10 min and embedded in a collagen-I gel before immunofluorescence, as described previously (13 (link)). Images were acquired with a SpinningDisk CSU-W1 (Yokogawa) with a Zyla sCMOC camera driven by an Olympus X83.

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Quantitative Fluorescence Microscopy of Centromeres

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Images were acquired on a Fluorescent microscope DeltaVision Core system (Applied Precision) with 100X Olympus UPlanSApo 100 oil-immersion objective (NA 1.4), 250W Xenon light source equipped with a Photometrics CoolSNAP_HQ2 Camera. ~ 4µm Z-stacks were acquired (Z step size: 0.2 µm). Imaris software (Bitplane) was used to quantify fluorescence intensity in the deconvolved 3D images using centromere surfaces automatically determined by ACA staining detection. Quantification of structured illumination (SI) images at centromeres was performed using Imaris software (Bitplane) surface fitting function and extracting data on each centromere volume and sphericity. All images presented were imported and processed in Photoshop (Adobe Systems, Inc S.J.). Movies of live cells were acquired using an Inverted Eclipse Ti-2 (Nikon) full motorized + Spinning disk CSU-W1 (Yokogawa) microscope. Cells were grown on high optical quality plastic slides (ibidi) for this purpose.

Giunta S., Hervé S., White R.R., Wilhelm T., Dumont M., Scelfo A., Gamba R., Wong C.K., Rancati G., Smogorzewska A., Funabiki H., & Fachinetti D. (2020). CENP-A chromatin prevents replication stress at centromeres to avoid structural aneuploidy.

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Quantifying Cellular Oxidation States

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HyPer fluorescence was excited with DPSS 10 mW 488 nm and 10 mW 405 nm lasers, and the corresponding YFP emission was measured using a 525/50 bandpass emission filter.
Spinning-disk images were acquired using a 63× objective (63×/1.4 oil WD: 0.17 mm) on a Spinning-Disk CSU-W1 (Yokogawa) equipped with a sCMOS Hamamatsu 2048×2048 camera. To calculate the HyPer ratio, images were treated as previously described (Mishina et al., 2013) .

Thauvin M., Amblard I., Rampon C., Mourton A., Quéguiner I., Li C., Gautier A., Joliot A., Volovitch M., & Vriz S. (2021). Reciprocal regulation of Shh trafficking and H₂O₂ levels via a noncanonical BOC-Rac1 pathway.

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Calcium Imaging of Colonic Muscles

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Colonic muscles obtained from PDGFRα-Cre-GCaMP6f mice were
equilibrated with continuous perfusion of warmed KRB solution at 37°C for
1 h. Calcium Imaging was performed using a spinning-disk confocal microscopy
(CSU-W1 spinning disk; Yokogawa Electric Corporation) mounted to an upright
Nikon Eclipse FN1 microscope equipped with a 60× 1.0 NA CFI Fluor lens
(Nikon instruments INC, NY, USA). GCaMP6f, expressed in PDGFRα cells
within colonic muscles excited at 488 nm using a laser coupled to a Borealis
system (ANDOR Technology, Belfast, UK). Emitted fluorescence (>515 nm)
was captured using a high-speed EMCCD Camera (Andor iXon Ultra; ANDOR
Technology, Belfast, UK). Image sequences were acquired at 33 fps using
MetaMorph software (Molecular Devices Inc., CA, USA) as previously described
(21 ). All experiments were performed
in the presence of atropine, L-NNA and MRS2500 to exclude effects from
cholinergic, nitrergic and purinergic pathways.

Kurahashi M., Kito Y., Baker S., Jennings L.K., Dowers J.G., Koh S.D, & Sanders K.M. (2020). A novel post-synaptic signal pathway of sympathetic neural regulation of murine colonic motility. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(4), 5563-5577.

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Structured Illumination Microscopy for Live-Cell Imaging

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Data shown in Figs. 1 and 5 were acquired on a Zyla 4.2 sCMOS camera (Andor) with a CSU-W1 spinning disk (Yokogawa) mounted on a Ti2 eclipse microscope (Nikon). The Apo Plan 100 × oil/NA1.4 and 60 × oil/NA1.4 objectives and a 405/488/561/638 nm laser (Omicron) were used in combination with an Okolab stage top incubation chamber (Okolab) in case of live-cell experiments. For SIM (3D structured illumination microscopy) images shown in Figs. 3 and 4, an N-SIM E (Nikon) was used, built on a Ti-Eclipse microscope (Nikon). Data were acquired using a z Piezo drive (Mad city labs), an Apochromat TIRF 100 × Oil/NA 1.49 objective, an Orca flash 4.0 LT sCMOS camera (Hamamatsu), a LU-N3-SIM 488/561/640 laser unit (Nikon) and a motorized N-SIM quad band filter combined with a single 525/50 emission filter using the laser line 488 at maximum output power. Z-stacks were acquired with a step size of 200 nm. Both microscopes were controlled by NIS-Elements software (Nikon). Slice reconstruction (NIS-Elements, Nikon) was performed using reconstruction parameters IMC 0.7, HNS 0.7, OBS 0.2.

Kirchenwitz M., Stahnke S., Grunau K., Melcher L., van Ham M., Rottner K., Steffen A, & Stradal T.E. (2022). The autophagy inducer SMER28 attenuates microtubule dynamics mediating neuroprotection. Scientific Reports, 12, 17805.

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Super-resolution Imaging Protocols

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For single-molecule localization super-resolution imaging, a TIRF microscope (Olympus IX81, Zero Drift Focus Compensator, Dual camera Hamamatsu ORCA-Fusion BT, Objective 100x) was used. PALM imaging of mEos2 and tdEos constructs was performed in PBS. Low intensity illumination at 405 nm was used to activate the fluorophores and 488 or 561 nm laser was used to image (and bleach) the activated fluorophores. Imaging was performed at 100 ms exposure for 7,000–10,000 frames.
SR-confocal imaging was performed using a spinning-disc confocal microscope based on a CSU-W1 Spinning Disk (Yokogawa) with a single, 70-μm pinhole disk and quad dichroic mirror along with an electron-multiplying charge-coupled device (EMCCD) (Princeton Instruments, ProEM HS 1024BX3 megapixels with 30-MHz cascade and eXcelon3 coating) using a Plan Apo 100× NA 1.45 objective lens. The LiveSR module (Roper Scientific France) was used to provide a structured illumination re-scanner that enhance spatial resolution ~2× by optical deconvolution.

Jain K., Minhaj R.F., Kanchanawong P., Sheetz M.P, & Changede R. (2024). Nano-clusters of ligand-activated integrins organize immobile, signalling active, nano-clusters of phosphorylated FAK required for mechanosignaling in focal adhesions. bioRxiv.

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Indirect Immunofluorescence Imaging of A549 Cells

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For indirect immunofluorescence analyses, 1.5 × 10⁵ A549 cells were grown on glass coverslips in 6-well cell dishes one day prior to infection. Cells were fixed with paraformaldehyde (PFA; 4% (v/v) in PBS) at room temperature (RT) for 20 min. PFA-fixed cells were permeabilized with phosphate-buffered saline (PBS) supplemented with 0.5% Triton X-100 for 10 min at room temperature and blocked with 1.5 mL Tris-buffered saline-BG (TBS-BG; BG represents 5% (w/v) BSA and 5% (w/v) glycine) for 1 h at RT. Next, coverslips were incubated with the indicated primary antibody diluted in PBS in a humidity chamber for 30 min. The corresponding secondary antibody (Alexa 488- (Invitrogen), Cy3 (Jackson, West Grove, PA, USA)-conjugated secondary antibodies) diluted in PBS was added afterwards. Finally, nuclei were stained with DAPI (4,6-diaminidino-2-phenylindole) in PBS (1:1000, (v/v) from 1 mg/mL stock) for 5 min, before the cover slips were mounted in mounting solution (Energene, Regensburg, Germany). Images were acquired using a confocal spinning-disk microscope (Nikon Eclipse Ti-E stand (Nikon, Tokyo, Japan); Yokogawa CSU-W1 spinning disk (Yokogawa, Tokyo, Japan; 2× Andor888 EM-CCD camera (Oxford Instruments, Abingdon, UK); Nikon 100× NA 1.49 objective (Nikon)).

Ip W.H., Wilkens B., Solomatina A., Martin J., Melling M., Hidalgo P., Bertzbach L.D., Speiseder T, & Dobner T. (2021). Differential Regulation of Cellular FAM111B by Human Adenovirus C Type 5 E1 Oncogenes. Viruses, 13(6), 1015.

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