Live imaging was performed on an Olympus IX70 microscope equipped with PerkinElmer Ultraview Vox confocal system or a Leica DM 1RB inverted microscope equipped with a Yokogawa CSU10 spinning disk confocal unit. All experiments were performed at room temperature with 63× or 100× oil objective. Eleven Z-stacks each 1 μm apart spanning the apical part of the nuclei were recorded every 30 to 120 sec, depending on the duration of time-lapse. GFP and RFP proteins were excited using a 488 nm and a 561 nm laser respectively. Data were acquired using Volocity 6 software (Quorum Technologies). Because nuclei move slightly inward into the interior of the embryos during cellularization in NC14, focal planes were manually adjusted every 15 to 30 min between time points when imaging embryos in NC14. Images being compared were acquired and processed using identical settings unless otherwise noted.
Csu10 spinning disk confocal unit
Manufactured by Yokogawa
The CSU10 is a spinning-disk confocal unit designed for high-speed, high-resolution imaging applications. It features a high-speed spinning Nipkow disk that enables rapid image acquisition, making it suitable for live-cell imaging and other dynamic processes. The CSU10 provides an efficient optical design for confocal fluorescence microscopy, allowing for the simultaneous capture of multiple wavelengths.
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Lab products found in correlation
3 protocols using csu10 spinning disk confocal unit
1
Live Imaging of Cellularization in Drosophila
2
Confocal Microscopy Imaging of Autophagy Markers
3
Confocal Microscopy Imaging of Autophagy Markers
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Immunofluorescence imaging was performed using the 100× (1.3 NA) objective of a Zeiss Axiovert 200 microscope equipped with a Spot RT camera (Diagnostics Instruments) and mercury lamp; images were acquired using Metamorph software (Molecular Devices v6.0). Confocal analysis of mCherry-GFP-LC3 puncta in reconstituted atg3−/− MEFs was performed using the 60× (1.4 NA) objective of a Nikon C1si spectral confocal system equipped with an argon laser (488 line) and two solid-state diodes (405 and 546 lines); images were acquired using Nikon EZ-C1 software. All other confocal analysis was performed using the 100× (1.49 NA) objective of a Nikon inverted microscope (TE-2000 PFS) equipped with a CSU10 spinning-disk confocal unit (Yokogawa), solid-state 488 and 561 lasers, and a cooled charge-coupled device camera (Cool-SNAP-HQ2, Photometrics); images were acquired using NIS Elements software. Images were analysed in ImageJ (v1.44i).
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