C. elegans were anesthetized with 0.1 mmol/L levamisole in M9 buffer for live-cell imaging of meiosis and germline oocyte. We synchronized young adult worms to image early embryos, cut them with syringes to release embryos in M9 buffer, and then mounted them on 2% (wt/vol) agarose pads at 20°C [31 (link)], time-lapse images were taken by μManager (www.micro-manager.org ) with an exposure time of 300 ms every 30–60 s. Our imaging system includes an Axio Observer Z1 microscope (Carl Zeiss MicroImaging, Inc.) equipped with a 100X, 1.45 NA objective, an EM CCD camera (Andor iXon+ DU-897D-C00-#BV-500), and the 488-nm and 561-nm lines of a Sapphire CW CDRH USB Laser System to a spinning disk confocal scan head (Yokogawa CSU-X1 Spinning Disk Unit). ImageJ software (NIH) was used to process and quantify the images (http://rsbweb.nih.gov/ij/ ). The background fluorescence was subtracted for the intensity measurements of UNC-85 and HIS-74 distribution. The whole-cell and nucleus fluorescence intensity of UNC-85 and HIS-74 were measured, and the nucleus to cytoplasm fluorescence ratio (N/C ratio) was calculated to reflect the distribution of UNC-85 and HIS-74.
Sapphire cw cdrh usb laser system
Manufactured by Yokogawa
Sourced in Germany
The Sapphire CW CDRH USB Laser System is a compact, continuous-wave (CW) laser solution that provides a stable, high-quality output beam. The system is designed to meet CDRH (Center for Devices and Radiological Health) safety requirements and can be controlled via a USB interface.
Automatically generated - may contain errors
Lab products found in correlation
Csu x1 spinning disk unit, by Yokogawa (9 mentions)
Axio observer z1 microscope, by Zeiss (9 mentions)
Ixon du 897d c00 bv 500, by Oxford Instruments (5 mentions)
Axio observer z1, by Zeiss (3 mentions)
Lsm 900 confocal microscope, by Zeiss (3 mentions)
Ix83 microscope, by Olympus (2 mentions)
Axio imager m2 microscope, by Zeiss (2 mentions)
Neo 5.5 scmos camera dc 152q c00 fi, by Oxford Instruments (1 mentions)
Emccd camera, by Oxford Instruments (1 mentions)
Dioc6 3, by Thermo Fisher Scientific (1 mentions)
10 protocols using sapphire cw cdrh usb laser system
1
Live-Cell Imaging of C. elegans Meiosis
2
Live-cell imaging of C. elegans
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To maximize the efficiency of live-cell imaging, we synchronized C. elegans by collecting ~100 young adult worms on the NGM plate to lay eggs for 2 h and then removing the adults. The eggs were maintained at 20 °C for ~16 h to develop into L1 larvae.
For live-cell imaging, L1 worms were anesthetized with 0.1 mmol/L levamisole in M9 buffer and mounted on 3% agarose pads at 20 °C. Live-cell imaging was performed using an Axio Observer Z1 microscope (Carl Zeiss MicroImaging, Inc.) equipped with a 100×, 1.45 N.A. objective, an EM CCD camera (Andor iXon+ DU-897D-C00-#BV-500), and the 488-nm and 568-nm lines of a Sapphire CW CDRH USB Laser System attached to a spinning disk confocal scan head (Yokogawa CSU-X1 Spinning Disk Unit). Time-lapse images were acquired with an exposure time of 200 ms every 30 s using the μManager (https://www.micro-manager.org/ ) and processed with ImageJ software.
For live-cell imaging, L1 worms were anesthetized with 0.1 mmol/L levamisole in M9 buffer and mounted on 3% agarose pads at 20 °C. Live-cell imaging was performed using an Axio Observer Z1 microscope (Carl Zeiss MicroImaging, Inc.) equipped with a 100×, 1.45 N.A. objective, an EM CCD camera (Andor iXon+ DU-897D-C00-#BV-500), and the 488-nm and 568-nm lines of a Sapphire CW CDRH USB Laser System attached to a spinning disk confocal scan head (Yokogawa CSU-X1 Spinning Disk Unit). Time-lapse images were acquired with an exposure time of 200 ms every 30 s using the μManager (
3
Quantitative Imaging of Microalgae and Nematodes
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Chlamydomonas cells were fixed with 0.5% Lugol’s solution at room temperature and imaged with an Axio Observer Z1 microscope (Carl Zeiss MicroImaging) equipped with a 100× 1.45 numerical aperture (NA) objective and an EM charge coupled device(CCD) camera (Andor iXon + DU-897DC00-# BV-500). Images were acquired with an exposure time of 200 ms by μManager (https://micro-manager.org/ ) and processed with ImageJ software. At least 100 cells were counted for each condition in biological triplicates.
Young-adult C. elegans hermaphrodites were anesthetized with 0.1 mmol/L levamisole, mounted on 3% agarose pads at 20 °C, and imaged with an Axio Observer Z1 microscope (Carl Zeiss MicroImaging) equipped with a 100× 1.45 NA objective, an EM CCD camera (Andor iXon + DU-897DC00-# BV-500), and the 488- and 568-nm lines of a Sapphire CW CDRH USB Laser System with a spinning-disk confocal scan head (Yokogawa CSU-X1 Spinning Disk Unit). Time-lapse images were acquired with an exposure time of 200 ms at 0-s intervals by μManager and processed with ImageJ software. The identical parameters were used to image the WT and mutant animals.
Young-adult C. elegans hermaphrodites were anesthetized with 0.1 mmol/L levamisole, mounted on 3% agarose pads at 20 °C, and imaged with an Axio Observer Z1 microscope (Carl Zeiss MicroImaging) equipped with a 100× 1.45 NA objective, an EM CCD camera (Andor iXon + DU-897DC00-# BV-500), and the 488- and 568-nm lines of a Sapphire CW CDRH USB Laser System with a spinning-disk confocal scan head (Yokogawa CSU-X1 Spinning Disk Unit). Time-lapse images were acquired with an exposure time of 200 ms at 0-s intervals by μManager and processed with ImageJ software. The identical parameters were used to image the WT and mutant animals.
4
Live-Cell Imaging of Worm Microvilli Dynamics
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Live-cell imaging of the worm was performed as described (43 (link)). Worms were anesthetized with 1 mg/mL levamisole and mounted on 3% agarose pads at 20 °C. The immobilized larvae were subsequently imaged with an Axio Observer Z1 microscope (Carl Zeiss MicroImaging, Inc.) and the 488 nm and 568 nm lines of a Sapphire CW CDRH USB Laser System attached to the spinning disk confocal scan head (Yokogawa CSU-X1 Spinning Disk Unit). We conducted live-cell imaging to follow microvilli dynamics at a high spatial resolution using a Zeiss LSM 900 confocal microscope with AiryScan superresolution module equipped with highly sensitive gallium arsenide phosphide detectors using a 63x Zeiss objective (1.4 numerical aperture) 20 °C.
5
High-Resolution Live Imaging of Worm Cells
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Young-adult worms were anesthetized with 0.1 mmol/L levamisole in M9 buffer and mounted on 3% agarose pads at 20°C. Live-cell imaging was performed on an Axio Observer Z1 microscope (Carl Zeiss, Oberkochen, Germany) equipped with a 100×, 1.49 numerical aperture (NA) objective, an electron-multiplying (EM) charge-coupled device (CCD) camera (Andor iXon+ DU-897D-C00-#BV-500), and the 488 nm and 561 nm lines of a Sapphire CW CDRH USB Laser System attached to a spinning disk confocal scan head (Yokogawa CSU-X1 Spinning Disk Unit). Our high-resolution live imaging system includes an Olympus IX83 microscope equipped with a 150×, 1.45 NA objective lens, a Neo 5.5 sCMOS Camera (DC-152Q-C00-FI; Andor Technology), and the same spinning disk confocal modules as mentioned above. Time-lapse images were acquired by μManager (https://www.micro-manager.org ) at an exposure time of 200 ms. Images of spectrin periodicity were collected on a Nikon (Tokyo, Japan) A1R laser-scanning confocal microscope with a CFI Plan Apo 100× oil immersion objective (NA 1.45) and 488-nm lasers with 150 nm X-Y resolution or a Hessian matrix-based structure illumination microscopy with 88 nm X-Y resolution.
6
Live Imaging of Fluorescent Proteins in Young Adults
7
Flagella Staining in Chlamydomonas
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Flagella were stained using a previously described protocol (46 (link)) with a few modifications. The algae cells were grown in TAP medium until the cell density reached ∼5 × 106 cell/mL and they were harvested and resuspended in TAP medium to a final cell concentration of ∼1 × 107 cell/mL Chlamydomonas cells were labeled with Dioc6(3) (0.05 μg/mL, Invitrogen) for 1 min at room temperature and imaged with an Axio Observer Z1 microscope (Carl Zeiss MicroImaging) equipped with the 488- and 568-nm lines of a Sapphire CW CDRH USB Laser System with a spinning-disk confocal scan head (CSU-X1 Spinning Disk Unit, Yokogawa).
8
Imaging Dendritic Arbors in Caenorhabditis elegans
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Young adult animals were anesthetized with 1 mg/ml levamisole in M9 buffer before mounted on 3% (w/v) agar pads. Images of the mec-3::GFP and the three corresponding independent eel-1 KO worms were captured in live animals at the same exposure time (2000 ms) using a 63× objective by AxioImager M2 microscope (Carl Zeiss). And images of PVD dendritic arbors of the related animals were captured by the spinning-disk confocal imaging system which includes an Axio Observer Z1 microscope (Carl Zeiss MicroImaging) equipped with a 40× objective, an electron-multiplying charge-coupled device camera (Andor), and the 488- and 568- nm lines of a Sapphire CW CDRH USB Laser System attached to a spinning-disk confocal scan head (Yokogawa CSU-X1 Spinning Disk Unit). Micro-Manager (https://micro-manager.org ) software and ImageJ (http://rsbweb.nih.gov/ij/ ) software were used to process the images.
9
Visualizing C. elegans Embryos
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C. elegans hermaphrodites were grown overnight in the dark at 20 °C on NGM plates containing 5 μM TMRE. Hermaphrodites were dissected and embryos were released from the gonads. Embryos were visualized using an Olympus IX83 microscope (Olympus Corp.), which is equipped with a 150 × , 1.45 N.A. objective, an EM CCD camera (Andor iXon+ DU-897D-C00-#BV-500), and the 405, 488 and 561 nm lines of a Sapphire CW CDRH USB Laser System attached to a spinning disk confocal scan head (Yokogawa CSU-X1 Spinning Disk Unit).
10
Imaging Subcellular Dynamics in C. elegans
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Q cell migration imaging was performed using the previously described protocol (32). L1 worms were anesthetized with 0.1 mmol/L levamisole in M9 buffer and mounted on 3% agarose pads at 20℃, then imaged on an Axio Observer Z1 microscope (Carl Zeiss) equipped with a 100×, 1.49 numerical aperture (NA) objective, an electron-multiplying (EM) charge-coupled device (CCD) camera (Andor iXon+ DU-897D-C00-#BV-500), and the 488 nm and 561 nm lines of a Sapphire CW CDRH USB Laser System attached to a spinning disk confocal scan head (Yokogawa CSU-X1 Spinning Disk Unit). Time-lapse images were acquired by μManager (https://www.micro-manager.org) at an exposure time of 200 msec. Images of spectrin and UNC-44 periodicity were collected on a Nikon (Tokyo, Japan) A1R laser-scanning confocal microscope with a CFI Plan Apo 100× oil immersion objective (numerical aperture 1.45) and 488-nm lasers with 150 nm X-Y resolution.
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