Fixed or living cells were observed at 25°C or 37°C with 5% CO2, respectively, under a TE2000 microscope (Nikon) equipped with a 100× 1.4 NA Plan-Apochromatic, a 60× 1.4 NA Plan-Apochromatic, or a 40× 1.3 NA Plan Fluor oil immersion objective lens (Nikon), a CSU-X1 confocal unit (Yokogawa), and an iXon3 electron multiplier-charge–coupled device camera (Andor) or an ORCA-ER CCD camera (Hamamatsu Photonics). Image acquisition was controlled by µManager software (Open Imaging). Long-term live imaging for cell cycle analyses was performed using a LCV110 microscope (Olympus) equipped with a 40× 0.95 NA UPLSAPO dry lens (Olympus). Because we found that 488-nm light irradiation severely interfered the progression of cell cycle and mitosis in HAP1 cells, we used bright-field microscope for long-term live imaging for cell cycle analyses (Figs. 1 , 8 , and S3). For live imaging, cells were cultured in phenol red–free IMDM (Thermo Fisher Scientific) supplemented with 10% FBS and 1× AA.
Csu x1 confocal unit
Manufactured by Yokogawa
Sourced in Japan
The CSU-X1 is a confocal unit designed for Yokogawa's range of microscopes. It employs a Nipkow spinning disk to provide optical sectioning capabilities for fluorescence imaging. The device utilizes a 405nm laser source and can capture images at up to 2000 frames per second.
Automatically generated - may contain errors
Lab products found in correlation
Te2000 microscope, by Nikon (6 mentions)
Ixon3 electron multiplier charge coupled device camera, by Oxford Instruments (4 mentions)
Orca er ccd camera, by Hamamatsu Photonics (3 mentions)
Plan apochromatic, by Nikon (3 mentions)
Plan fluor oil immersion objective lens, by Nikon (3 mentions)
Zyla 4.2 scmos camera, by Oxford Instruments (3 mentions)
Ti2 microscope, by Nikon (3 mentions)
Phenol red free imdm, by Thermo Fisher Scientific (2 mentions)
Plan apochromatic objective lens, by Nikon (2 mentions)
Sir tubulin, by Cytoskeleton (2 mentions)
Uplansapo, by Olympus (2 mentions)
Lcv110 microscope, by Olympus (1 mentions)
1.4na oil immersion objective lens, by Nikon (1 mentions)
Metamorph software, by Molecular Devices (1 mentions)
Emccd camera, by Oxford Instruments (1 mentions)
8 protocols using csu x1 confocal unit
1
Live Cell Imaging of Cell Cycle
2
Live-cell and Electron Microscopy Imaging
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The fixed and living cells were observed under a TE2000 microscope (Nikon, Tokyo, Japan) equipped with a 100×/1.4 numerical aperture (NA) Plan-Apochromatic, a 60×/1.4 NA Plan-Apochromatic, or a 40×/1.3 NA Plan Fluor oil immersion objective lens (Nikon), a CSU-X1 confocal unit (Yokogawa, Tokyo, Japan), and an iXon3 electron-multiplier charge coupled device camera (Andor, Belfast, United Kingdom). Image acquisition was controlled by μManager software. Fluorescence recovery after photobleaching was performed using a C2 (Nikon) confocal microscope equipped with a 60×/1.4 NA oil immersion objective lens (Nikon). TEM was performed as previously described (Uehara et al., 2013 (link)).
3
Live Cell Imaging of Microtubules
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Fixed cells were observed at 25°C under a TE2000 microscope (Nikon) equipped with a × 100 1.4 numerical aperture (NA) Plan-Apochromatic objective lens (Nikon), a CSU-X1 confocal unit (Yokogawa), and an iXon3 electron multiplier-charge coupled device camera (Andor). Living cells were observed for 24 h at 37°C with 5% CO2 under a Ti-2 microscope (Nikon) equipped with a × 40 0.95 NA Plan-Apochromatic objective lens (Nikon), and Zyla4.2 sCMOS camera (Andor). Sir-tubulin (Cytoskeleton, Inc.) was treated at 250 nM for live imaging. Image acquisition was controlled by μManager (Open Imaging).
4
Confocal Microscopy of Yeast Cells
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Logarithmically growing yeast cells cultivated in SCD were observed using a model IX83 microscope (Olympus) with a CSU-X1 confocal unit (Yokogawa), a 100× and 1.4 numerical aperture objective lens (UPlanSApo; Olympus), and an EM-CCD camera (Evolve 512; Photometrics). MetaMorph software (Molecular Devices) was used to analyze images. GFP was excited using a 488-nm laser (OBIS; Coherent) and the emission was passed through 520/35-nm filter. The confocal fluorescent sections were collected every 0.2 μm from the upper to the bottom surface of yeast cells. The obtained confocal images were subjected to maximum projection using Image J software (NIH).
5
Microscopic Imaging of Cellular Dynamics
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Cells were observed under a TE2000 microscope (Nikon) equipped with a × 100 1.4 numerical aperture (NA) Plan-Apochromatic, a × 60 1.4 NA Plan-Apochromatic, or a × 40 1.3 NA Plan Fluor oil immersion objective lens (Nikon), a CSU-X1 confocal unit (Yokogawa), and an iXon3 electron multiplier-charge coupled device (EMCCD) camera (Andor) or an ORCA-ER CCD camera (Hamamatsu Photonics). Image acquisition was controlled by µManager software (Open Imaging).
6
Confocal Microscopy of Fixed and Live Cells
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Fixed cells were observed under a TE2000 microscope (Nikon, Tokyo, Japan) equipped with a ×100 1.4 NA Plan‐Apochromatic, a CSU‐X1 confocal unit (Yokogawa, Tokyo, Japan) and an iXon3 electron multiplier‐charge coupled device (EMCCD) camera (Andor, Belfast, UK) or ORCA‐ER CCD camera (Hamamatsu Photonics, Tokyo, Japan), or with a Ti2 microscope (Nikon) with ×60 1.4 NA Apochromatic, and Zyla4.2 sCMOS camera (Andor). Live cell imaging was conducted at 37 °C with 5% CO2 using a Ti‐2 microscope with ×20 0.75 NA Plan‐Apochromatic and Zyla4.2. For live imaging, cells were cultured in phenol red‐free IMDM (Thermo Fisher Scientific) supplemented with 10% FBS and 1× AA. Image acquisition was controlled by μManager (an open‐source software program)[27 ].
7
Live Cell Imaging with Tubulin Labeling
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Fixed cells were observed at 25°C under a TE2000 microscope (Nikon) equipped with a ×100 1.4 numerical aperture (NA) Plan-Apochromatic objective lens (Nikon), a CSU-X1 confocal unit (Yokogawa), and an iXon3 electron multiplier-charge coupled device camera (Andor). Living cells were observed for 24 h at 37°C with 5% CO2 under a Ti-2 microscope (Nikon) equipped with a ×40 0.95 NA Plan-Apochromatic objective lens (Nikon), and Zyla4.2 sCMOS camera (Andor). Sir-tubulin (Cytoskeleton, Inc.) was treated at 250 nM for live imaging. Image acquisition was controlled by µManager (Open Imaging).
8
Imaging Yeast Peroxisomal Dynamics
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Yeast cells expressing 3xFLAG-GFP-Pex15D30 or 3xFLAG-GFP-Pex15KRD30 were grown in SCGalSuc or SCGAL medium supplemented with 20 mg/ml adenine to suppress background signal from vacuoles medium at 30 C. Yeast cells expressing the split GFP proteins were grown in SCD medium at 30 C. Cells grown at log-phase were observed under Olympus IX83 microscope with a CSU-X1 confocal unit (Yokogawa), a 100A ˚$, 1.4 NA, objective (UPlanSApo, Olympus) and an EM-CCD camera (Evolve512; Photometrics) manipulated by Metamorph software (Molecular Devices). GFP was excited by 488-nm (OBIS, Coherent) and the emission was passed through a 520/35-nm band-pass filter. RFP, mCherry and mScarlet were excited by 561-nm laser (OBIS, Coherent) and the emission was passed through a 617/73-nm band-pass filter. The confocal fluorescent sections were collected every 0.2 mm from the upper to bottom surface of yeast cells.
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