Csu w1 t3

Manufactured by Yokogawa

The CSU-W1-T3 is a laboratory equipment product manufactured by Yokogawa. It is a compact, stand-alone unit designed for temperature control applications. The device features a temperature range and control accuracy, but further details are not available without the risk of making unsubstantiated claims.

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Lab products found in correlation

Axio observer, by Zeiss (12 mentions) Axioobserver z1, by Yokogawa (2 mentions) Proem 1024, by Teledyne (2 mentions) Fm4 64 styryl dye, by Thermo Fisher Scientific (2 mentions) Lsm 710, by Zeiss (2 mentions) Plan apochromatic objective, by Zeiss (1 mentions) Sp8x microscope, by Leica camera (1 mentions) Lysotracker red dnd 99, by Thermo Fisher Scientific (1 mentions) Sic001, by Merck Group (1 mentions) Lsm 710 inverted confocal microscope, by Zeiss (1 mentions) Axio scan z1, by Zeiss (1 mentions) Plan apochromat objective, by Yokogawa (1 mentions) Prism 6, by GraphPad (1 mentions)

14 protocols using csu w1 t3

Neuron Imaging in Microfluidic Devices

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Videorecording of neurons plated in microfluidic devices was performed at DIV 12. Before recordings, DIV 12 neurons in the microchamber were carefully inspected and selected based on the absence of cell contamination. For double transductions (with sh-RNA lentiviruses), the transport of mCherry-tagged cargo was analyzed within GFP-positive axons. Images were acquired every 200 ms for 1 min on an inverted microscope (Axio Observer, Zeiss) with X63 oil-immersion objective (1.46NA) coupled to a spinning-disk confocal system (CSU-W1-T3; Yokogawa) connected to an electron-multiplying CCD (charge-coupled device) camera (ProEM+1024, Princeton Instrument) at 37°C and 5% CO₂.
For the study of the exocytosis events, images were acquired every 200 ms for 1 min on an inverted microscope (Axio Observer, Zeiss) with X63 oil-immersion objective (1.46NA) coupled to a spinning-disk confocal system (CSU-W1-T3; Yokogawa) with TIRF microscopy (Nikon/Roper, Eclipse Ti) equipped with a camera Prime 95B sCMOS (Telelyne Photometrics) at 37°C and 5% CO₂. The same three fields per microchambers were acquired before and after a 4AP-bicuculline (respectively 2.5 mM and 50 µM) stimulation of the presynaptic chamber, four times in total (one before and three after stimulation).

Vitet H., Bruyère J., Xu H., Séris C., Brocard J., Abada Y.S., Delatour B., Scaramuzzino C., Venance L, & Saudou F. (2023). Huntingtin recruits KIF1A to transport synaptic vesicle precursors along the mouse axon to support synaptic transmission and motor skill learning. eLife, 12, e81011.

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Multicolor Confocal Imaging of Arabidopsis Roots

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Images of the SAND lines were performed on an inverted Zeiss LSM710 confocal microscope using a 40x Plan-apochromatic objective (numerical aperture 1.2, oil immersion). Dual-colour images were acquired by sequential line switching, allowing the separation of channels by both excitation and emission. Counter staining of plasma membranes were obtained by incubating roots with 1 μM FM4-64 solution (Lifesciences, stock solution at 3mM in H₂O) for 10 minutes, followed by 15 to 45 minutes incubation in water prior to observation. mCitrine and FM4-64 were excited with a 515nm laser.
Images of the BREAK lines were performed on an inverted Zeiss microscope (AxioObserver Z1) equipped with a spinning disk module (CSU-W1-T3, Yokogawa) and a ProEM+ 1024B camera (Princeton Instrument) using a 40x C-Apochromat objective (numerical aperture 1.1, water immersion). GFP was excited with a 488nm laser.
Images of the RED TIDE and LINE UP lines, as well as irt1 complementation analyses were performed on an inverted Leica SP8 X microscope using a 40x HCX Plan-apochromatic objective (numerical aperture 1.3, oil immersion). Yellow fluorescent proteins (YFP, mCitrine and Ypet) and mCherry were excited with a white light laser at 514nm and 587 nm, respectively. Imaging of LINE UP lines was performed after induction with 30 μM dexamethasone (DEX) for 24 hours.

Marquès-Bueno M.D., Morao A.K., Cayrel A., Platre M.P., Barberon M., Caillieux E., Colot V., Jaillais Y., Roudier F, & Vert G. (2016). A versatile Multisite Gateway-compatible promoter and transgenic line collection for cell type-specific functional genomics in Arabidopsis. The Plant journal : for cell and molecular biology, 85(2), 320-333.

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Live Imaging of Lysosomal Dynamics

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Live imaging of KO cultures grown in MFCs was conducted on DIV12. Lysotracker Red DND-99 (Thermo Fisher Scientific, L7528) was added to the somatodendritic compartment of the MFC to a final dilution of 1:10,000. Lysotracker-stained vesicles became visible in the axonal compartment a few minutes later. Imaging was conducted at 37°C and 5% CO₂ on a Zeiss Axio Observer coupled to a spinning disk confocal system (CSU-W1-T3; Yokogawa) connected to an electron-multiplying CCD camera (ProEM+1024, Princeton Instruments). Images were taken every 1 s for 180 s with a 63×/1.46 NA oil immersion objective.

Konietzny A., Han Y., Popp Y., van Bommel B., Sharma A., Delagrange P., Arbez N., Moutin M.J., Peris L, & Mikhaylova M. (2024). Efficient axonal transport of endolysosomes relies on the balanced ratio of microtubule tyrosination and detyrosination. Journal of Cell Science, 137(8), jcs261737.

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Live-cell Trafficking Kinetics Characterization

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Live-cell recordings were performed using an inverted microscope (Axio Observer, Zeiss) coupled to a spinning-disk confocal system (CSU-W1-T3, Yokogawa) connected to wide field electron-multiplying CCD camera (ProEM+1024, Princeton Instrument) and maintained at 37°C and 5% CO2. For HTT-mCherry and BDNF-mCherry trafficking, images were taken every 200 ms for 30 s (63x oil-immersion objective, 1.46 NA). Kymographs were generated using KymoToolBox plugin for ImageJ (Zala et al., 2013 (link)) to extract the following kinetics parameters (previously described in Virlogeux et al., 2018 (link)): anterograde/retrograde velocity, number of anterograde/retrograde vesicles per 100 μm, linear flow rate, directional flux. For each experiment, at least 2-3 microchambers from 3 independent neuronal cultures were used, and a minimum number of 50-60 axons were analyzed.

Migazzi A., Scaramuzzino C., Anderson E.N., Tripathy D., Hernández I.H., Grant R.A., Roccuzzo M., Tosatto L., Virlogeux A., Zuccato C., Caricasole A., Ratovitski T., Ross C.A., Pandey U.B., Lucas J.J., Saudou F., Pennuto M, & Basso M. (2021). Huntingtin-mediated axonal transport requires arginine methylation by PRMT6. Cell reports, 35(2), 108980.

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Golgi dynamics in STHdh cell lines

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STHdhQ7/Q7 and STHdhQ111/Q111 were electroporated with the KDEL-GPI-mCherry plasmid (gift of F. Perez) and when indicated with si-CT (Sigma-Aldrich, SIC001), si-APT1 (Sigma-Aldrich, SASI_Mm01_00077477), and si-APT2 (Sigma-Aldrich, SASI_Mm01_00051133) using the cell line nucleofactor kit L (Lonza) according to the manufacturer’s instructions. STHdh cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum, 1% of nonessential amino acids, 2 mM l-glutamine, and at 33°C in a humidified 5% CO₂ atmosphere. After adding 40 μM biotin, images were taken every 30 s over a 90-min period using an inverted microscope (Axio Observer, Zeiss) coupled to a spinning-disk confocal system (CSU-W1-T3, Yokogawa) connected to a wide-field electron-multiplying charge-coupled device (CCD) camera (ProEM⁺1024, Princeton Instrument) [63× oil-immersion objective, 1.46 numerical aperture (NA)] and maintained at 33°C and 5% CO₂. Integrated fluorescence intensity was measured in the Golgi region, corrected for background and normalized to the maximum value.

Virlogeux A., Scaramuzzino C., Lenoir S., Carpentier R., Louessard M., Genoux A., Lino P., Hinckelmann M.V., Perrier A.L., Humbert S, & Saudou F. (2021). Increasing brain palmitoylation rescues behavior and neuropathology in Huntington disease mice. Science Advances, 7(14), eabb0799.

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Microscopy Imaging of Neuronal Signaling

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All the live cell images were acquired using an inverted microscope (Axio Observer, Zeiss) coupled to a spinning disk confocal system (CSU-W1-T3; Yokogawa) connected to an electron-multiplying CCD (charge-coupled device) camera (ProEM+1024, Princeton Instrument) and maintained at 37°C and 5% CO₂. Images were taken every 0.2 s for 30 s for TrkB-mCherry, TrkA-GFP, and EGFR-GFP trafficking and for the dual color acquisition (488 to 565) of TrkB-GFP and QDs-BDNF [with a 63× 1.46 numerical aperture (NA) oil immersion objective]. TrkB, HTT, and CaN immunostaining were visualized inside the cortical axonal microchannel, whereas for p-TrkB and PSD95, the Z-stack acquisitions were taken in the cortical layer V of brain slices. Immunostaining images were acquired with a 63× oil immersion objective (1.4 NA) using a Zeiss LSM 710 inverted confocal microscope coupled to an Airyscan detector to improve signal-to-noise ratio and resolution. For STED microscopy, we imaged with a 100× oil immersion objective (1.46 NA) using the Abberior 2D-STEDYCON upright confocal microscope. Phospho-ERK immunohistochemistry was visualized with a 20× objective (0.45 NA) using a slide scanner (AxioScan Z1, Zeiss).

Scaramuzzino C., Cuoc E.C., Pla P., Humbert S, & Saudou F. (2022). Calcineurin and huntingtin form a calcium-sensing machinery that directs neurotrophic signals to the nucleus. Science Advances, 8(1), eabj8812.

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Visualizing Synaptic Vesicle Dynamics

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Neurons were seeded into microfluidic devices and transduced at DIV1 with GFP lentivirus. The synaptic chamber was incubated at indicated time with 10 µM of FM4-64 styryl dye (ThermoFischer Scientific) into high KCl Tyrode solution (2 mM NaCl; 50 mM KCl; 2 mM CaCl₂; 1 mM MgCl₂; 10 mM Glucose and 1 mM Hepes buffer pH7.4) during 1 min at 37°C. After three washes with Tyrode solution (150 mM NaCl; 4 mM KCl; 2 mM CaCl₂; 1 mM MgCl₂; 10 mM Glucose and 1 mM Hepes buffer pH7.4) containing inhibitors of additional firing (1 mM kynurenic acid and 10 mM MgCl₂), acquisitions were made on inverted microscope (Axio Observer, Zeiss) coupled to a spinning-disk confocal system (CSU-W1-T3; Yokogawa) connected to an electron-multiplying CCD (charge-coupled device) camera (ProEM+1024, Princeton Instrument) at 37°C and 5% CO2 with z stacks of 5 µm.

Bruyère J., Abada Y.S., Vitet H., Fontaine G., Deloulme J.C., Cès A., Denarier E., Pernet-Gallay K., Andrieux A., Humbert S., Potier M.C., Delatour B, & Saudou F. (2020). Presynaptic APP levels and synaptic homeostasis are regulated by Akt phosphorylation of huntingtin. eLife, 9, e56371.

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Pollen Tube Growth Kinetics

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Flowers at stage 13 and 15 were emasculated and pollinated on plants with mature pollen from the pACT11::RFP line. Immediately after pollination, stigmas were mounted between two coverslips maintained separated by four grease plugs placed at each coverslip corner. To maintain a constant humidity without adding liquid directly on the stigma surface, we used a wet piece of tissue in contact with the base of the stigma. Pollinated stigmas were observed under a Zeiss microscope (AxioObserver Z1) equipped with a spinning disk module (CSU-W1-T3, Yokogawa) using a 40x Plan-Apochromat objective (numerical aperture 1.1, water immersion). Serial confocal images were acquired in the entire volume of the stigma every 1 µm and every minute. Images were processed with Image J software and pollen tube lengths were measured.

Riglet L., Rozier F., Kodera C., Bovio S., Sechet J., Fobis-Loisy I, & Gaude T. (2020). KATANIN-dependent mechanical properties of the stigmatic cell wall mediate the pollen tube path in Arabidopsis. eLife, 9, e57282.

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Imaging Axonal Transport of BDNF-mCherry

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In vivo axonal transport of BDNF-mCherry was imaged in cortical neurons plated in microfluidic chambers. Neuronal processes can grow into the microchannels, but only axons can reach the distal part of the micro channel located 450 μm away from the proximal chamber where cell bodies are located (Fig. 2c). Acquisitions were done in the distal part of the microchannels, using an inverted microscope (Axio Observer, Zeiss) with a × 63 1.46 numerical aperture (NA) oil-immersion objective coupled to a spinning-disk confocal system (CSU-W1-T3; Yokogawa) connected to an electron-multiplying CCD (charge-coupled device) camera (ProEM⁺1024, Princeton Instrument) and maintained at 37 °C and 5% CO₂. Images were taken every 0.2 s for 30 s. Kymographs and velocity analysis were done using KymoToolBox6 (link), a homemade plugin for ImageJ (NIH, USA). Briefly, calibrated kymographs were generated from maximal projection of time-lapse videos. Vesicle trajectories were then analysed manually, to extract the dynamic parameters of the tracked particles. Only moving vesicles were analysed; therefore, static vesicles or particles moving at velocities lower than 0.2 μm s⁻¹ were not taken into account.

Hinckelmann M.V., Virlogeux A., Niehage C., Poujol C., Choquet D., Hoflack B., Zala D, & Saudou F. (2016). Self-propelling vesicles define glycolysis as the minimal energy machinery for neuronal transport. Nature Communications, 7, 13233.

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Live-cell imaging of neuronal dynamics

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Live-cell videorecordings were performed using an inverted microscope (Axio Observer, Zeiss) coupled to a spinning-disk confocal system (CSU-W1-T3, Yokogawa) connected to wide-field electron-multiplying CCD camera (ProEM⁺ 1024, Princeton Instrument) and maintained at 37 °C and 5% CO₂. Images were taken at 5 Hz for 30 s for BDNF-mCh trafficking using a ×63 oil-immersion objective (1.46 NA), at 1 Hz for 5 min for Mito-DsRed2 using a ×63 oil-immersion objective (1.46 NA), and at 5 Hz for 30 s for GCaMP6f using a ×20 objective (0.8 NA). iGluSnFr was imaged live using z-stack acquisitions with a ×63 oil-immersion objective (1.46 NA) in the synaptic chamber. Fixed immunostaining images of SYP/PSD95 were obtained using z-stack acquisitions with a ×63 oil-immersion objective (1.4 NA) using an inverted confocal microscope (LSM 710, Zeiss) coupled to an Airyscan detector to improve signal-to-noise ratio and spatial resolution.

Moutaux E., Christaller W., Scaramuzzino C., Genoux A., Charlot B., Cazorla M, & Saudou F. (2018). Neuronal network maturation differently affects secretory vesicles and mitochondria transport in axons. Scientific Reports, 8, 13429.

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