For live imaging of L3 and young adult hermaphrodites, animals were sedated with 10 mM Levamisole M9 and mounted on 5% agarose. To perform imaging on embryos, gravid adults were dissected in M9 and mounted on 5% agarose. Two main microscopes were used. First, a wide-field Zeis Axioplan2 upright microscope equipped with 25x – 0.8 NA, 63x – 1.4 NA, and 100x – 1.4 NA objectives, and an Axiocam MRm CCD monochrome camera. Zeiss filter sets used were set 34 for DAPI, set 13 for GFP, and set 31 for mCherry. The microscope and camera were controlled by Zeiss Axiovision 4.x software. Second, an Andor spinning disc platform controlled by MetaMorph software and consisting of a Nikon Ti-U inverted microscope with 60x – 1.4 NA and 100x – 1.4 NA oil objectives, a Yokogawa CSU-X1 spinning disk unit, 488 nm and 561 nm lasers, Semrock 512 – 23 + 630 – 91, 525 – 30, and 617 – 73 emission filters, and an Andor iXON DU-885 camera. Images in Figure 6 were captured using a Zeiss AxioImager, 40x – 1.3 NA objective, and Hammamatsu Orca-R2 camera. Images of fixed embryos were deconvolved using AxioVision software, and are shown as maximum intensity projections of 3–5 adjacent planes spaced 0.3 μm apart. Images were cropped, rotated, and levels were adjusted in ImageJ and Adobe Photoshop.
Ixon du 885 camera
Manufactured by Oxford Instruments
Sourced in United Kingdom
The IXON DU-885 is a scientific camera designed for high-performance imaging applications. It features a large-format sensor, high quantum efficiency, and low noise characteristics, enabling it to capture detailed, low-light images with exceptional quality. The camera's core function is to provide researchers and scientists with a versatile imaging tool for a wide range of scientific and industrial applications.
Automatically generated - may contain errors
3 protocols using ixon du 885 camera
1
Live Imaging of C. elegans Embryos and Worms
2
Vinculin Clusters and Actin Cytoskeleton in PC12 Cells
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PC12 cells were seeded on the different substrates (on Ø24 mm glass cover slips). At the indicated time points (30 min, 1, 4 and 24 h) the cells were fixed and labeled for vinculin and f-actin following the immunofluorescence imaging protocol described above. The images were recorded with a Leica AM TIRF MC system using a Leica HCX PL APO 63X NA 1.47 objective (Leica). To visualize the vinculin clusters, integrated 488 nm laser lines and Andor iXon DU-885 camera (Andor Technology, Belfast, UK) was used. The image recording was done with a laser incident angle of 74° allowing a penetration depth of almost 250 nm. The images were elaborated with ImageJ following a recently described method [96 (link)] to analyze the area and number of clusters per cell. In total, for global statistics 722-3678 clusters from 16–34 cells were analyzed from three independent experiments. The f-actin instead was imaged in epifluorescence mode. Here the cytoskeletal organization of the cells was categorized in three categories: (1) no detectable presence of actin bundles/stress fibers, (2) 1–10 distinct actin bundles/stress fibers and (3) >10 distinct actin bundles/stress fibers.
3
Live Imaging of C. elegans Samples
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For live imaging of L3 and young adult hermaphrodites, animals were sedated with 10 mM Levamisole M9 and mounted on 5% agarose. To perform imaging on embryos, gravid adults were dissected in M9 and mounted on 5% agarose. Two main microscopes were used. First, a wide-field Zeis Axioplan2 upright microscope equipped with 25x − 0.8 NA, 63x − 1.4 NA, and 100x − 1.4 NA objectives, and an Axiocam MRm CCD monochrome camera. Zeiss filter sets used were set 34 for DAPI, set 13 for GFP, and set 31 for mCherry. The microscope and camera were controlled by Zeiss Axiovision 4.x software. Second, an Andor spinning disc platform controlled by MetaMorph software and consisting of a Nikon Ti-U inverted microscope with 60x − 1.4 NA and 100x − 1.4 NA oil objectives, a Yokogawa CSU-X1 spinning disk unit, 488 nm and 561 nm lasers, Semrock 512 − 23 + 630 − 91, 525 − 30, and 617 − 73 emission filters, and an Andor iXON DU-885 camera. Images in Figure 6 were captured using a Zeiss AxioImager, 40x − 1.3 NA objective, and Hammamatsu Orca-R2 camera. Images of fixed embryos were deconvolved using AxioVision software, and are shown as maximum intensity projections of 3–5 adjacent planes spaced 0.3 μm apart. Images were cropped, rotated, and levels were adjusted in ImageJ and Adobe Photoshop.
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