Spinning disk movies were acquired using a Plan APO 40 Å~/1.25 N.A. objective on a Leica DMI6000B microscope enclosed in a thermostatic chamber at 37 °C (Life Imaging Services), and equipped with a CoolSnap HQ2/CCD-camera (Princeton Instruments) or EMCCD camera (Evolve) coupled to a Sutter filter wheel (Roper Scientific) and a Yokogawa CSU-X1-M1 confocal scanner. MetaMorph software (Universal Imaging) collected the data.
Csu x1 m1
Manufactured by Yokogawa
The CSU-X1-M1 is a laboratory equipment product offered by Yokogawa. It serves as a core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Axio observer z1, by Zeiss (4 mentions)
Sutter filter wheel, by Teledyne (3 mentions)
Metamorph software, by Universal Imaging (3 mentions)
Coolsnap hq2 ccd camera, by Teledyne (2 mentions)
Fibronectin coated glass bottomed dishes, by MatTek (2 mentions)
Dmi6000, by Leica (1 mentions)
Ixon ultra 888 emccd camera, by Oxford Instruments (1 mentions)
Eclipse ti2 e, by Nikon (1 mentions)
7 protocols using csu x1 m1
1
Spinning Disk Microscopy for Live-Cell Imaging
2
Time-Lapse Imaging of Transfected Cells
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Cells were transfected, incubated for 24 hr, and re-plated prior to imaging on 10 μg/ml fibronectin–coated glass-bottomed dishes (MatTek Corporation). The time-lapse images were acquired with 3I Marianas imaging system (3I intelligent Imaging Innovations), with an inverted spinning disk confocal microscope Zeiss Axio Observer Z1 (Zeiss) and a Yokogawa CSU-X1 M1 confocal scanner, or with an inverted microscope (IX-71; Olympus) equipped with a Polychrome IV monochromator (TILL Photonics). Both systems have appropriate filters, heated sample chamber (+37°C), and controlled CO2. With 3I Marianas, a 63x/1.2 W C-Apochromat Corr WD = 0.28 M27 objective was used. SlideBook 5.0 software (3I intelligent Imaging Innovations) and sCMOS (Andor) Neo camera were used for the image acquirement and recording. With Olympus, a 60x water objective with 1.6× magnification was used. TILL Vision 4 software (TILL Photonics) and Imago QE (TILL Photonics) and Andor iXon (Andor) cameras were used for the image acquirement and recording. Deconvolution of the time-lapse videos was performed with AutoQuant AutoDeblur 2D non-blind Deconvolution (AutoQuant Imaging, Inc.). Further analyses of the video frames were performed with Image Pro Plus 6.0.
3
Live-cell Confocal Microscopy Imaging
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Spinning disk videos were acquired using a Plan APO 40×/1.25-NA objective on a Leica DMI6000B microscope, enclosed in a thermostatic chamber set at 37°C (Life Imaging Services) and equipped with a CoolSnap HQ2/CCD-camera (Princeton Instruments) or EMCCD camera (Evolve) coupled to a Sutter filter wheel (Roper Scientific) and a Yokogawa CSU-X1-M1 confocal scanner. MetaMorph software (Universal Imaging) was used to collect the data.
4
Spinning Disk Microscopy Imaging
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Spinning disk images and movies were acquired using a Plan-APO 40×/1.25 NA objective on a Leica DMI6000B microscope enclosed in a thermostatic chamber (Life Imaging Service) equipped with a Retiga 3 CCD camera (QImaging) coupled to a Sutter filter wheel (Roper Scientific) and a Yokogawa CSU-X1-M1 spinning disk. Metamorph software (Universal Imaging) was used to collect data.
5
Tracking Phagocytosis of HIV-1 Infected Cells
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To follow phagocytosis on HIV-1 GFP–infected or noninfected cells, images were recorded every min for 2 h on a spinning disk confocal (CSU-X1M1; Yokogawa) inverted microscope (DMI6000; Leica) equipped with a CoolSnap HQ2 camera (Photometrics) and a heated chamber and CO2 in the BSL3 laboratory. HIV-1–GFP and SRBCs were visualized by fluorescence and phase contrast with a 100×, 1.4 NA, PH differential interference contrast objective. Acquisition was performed with MetaMorph 7.5.5 software (Molecular Devices). The movies were analyzed using ImageJ with the Manual Tracking Plugin (F. Cordelières, Institut Curie, Orsay, France). Distances relative to the nucleus (visible on phase contrast images) were calculated in micrometers and the traveled distances were calculated and plotted against time. The slopes were calculated with linear regression.
6
Mitochondrial dynamics in U2OS cells
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U2OS cells were treated with FAM92A1 siRNA for 72 h, and the cells were replated on 10 µg/ml fibronectin-coated glass-bottomed dishes (MatTek) after 48 h transfection of mitochondrial matrix–targeted YFP (mito-YFP). The time-lapse images were acquired with a Marianas imaging system (3I) equipped with an inverted spinning-disk confocal microscope (AxioObserver Z1; Zeiss) and a Yokogawa CSU-X1 M1 confocal scanner. Cells were placed in heated sample chamber (37°C) and controlled CO2. A 63× 1.2 W C-Apochromat Corr working distance = 0.28 M27 objective was used, and all the images were acquired by an sCMOS (Andor) Neo camera and Slidebook 5.0 software (3I). Analyses of the video frames were performed with Image Pro Plus 7.0.
7
Chronic Live Imaging of Cellular Dynamics
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Live imaging was performed with a custom built confocal spinning disc setup built of an inverted microscope (EclipseTi2-E, Nikon Instruments), and a spinning disc device (CSU-X1-M1, Yokogawa). Images were acquired with a 488 and a 561 nm lasers (Sapphire, Coherent) and an iXon Ultra888 EMCCD camera (Andor,Oxford Instruments). Z-stacks were acquired with a z-interval of 1 . Laser power and exposure were kept as low as possible for chronic imaging, to reduce phototoxicity. The two color channels (GFP and mCherry, 488 and 564 nm lasers, respectively), where acquired in sequence. All images were obtained with a 60X water-immersion objective (Plan Apo 60×, NA 1.2, Nikon).
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