Freshly hatched medaka larvae containing yolk sac were used for in vivo imaging as described previously (20 (link)). Briefly, time-lapse in vivo recording the entire larval thymus were carried out on a PerkinElmer Ultraview VoX or Ultraview ERS Spinning disk confocal using a 40× water-immersion objective (LD C-Apochromat, 1.1 numerical aperture, Corr, Zeiss). For accurate tracking of individual cells, z-stacks of 60- to 70-μm spanning the whole thymus area (z-space, 1 μm) were imaged with a time interval of <15 s.
Ultraview ers
Manufactured by Zeiss
The Ultraview ERS is a high-performance fluorescence microscopy system developed by Zeiss. It is designed for advanced imaging applications in cell biology, neuroscience, and other life science research fields. The Ultraview ERS provides fast, high-resolution imaging of live and fixed samples with exceptional image quality.
Automatically generated - may contain errors
Lab products found in correlation
Ultraview vox, by PerkinElmer (1 mentions)
Axiovert 200m inverted microscope, by Zeiss (1 mentions)
Lsm 5 live, by Zeiss (1 mentions)
Mitotracker red cmxros, by Thermo Fisher Scientific (1 mentions)
Glass coverslip, by Corning (1 mentions)
Cox 4, by Cell Signaling Technology (1 mentions)
Cyclin b1 gns1, by SCBT (1 mentions)
Map lc3β g 9, by SCBT (1 mentions)
Axiovert 200, by Zeiss (1 mentions)
Vectashield hardset mounting medium with dapi, by Vector Laboratories (1 mentions)
4 protocols using ultraview ers
1
In Vivo Imaging of Medaka Thymus
2
Real-Time Tracking of Cell Migration
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Digitized bright-field time-lapse real-time images of the movement of MDA-MB231 cells (placed in live-cell imaging chambers) into the scratched area (for migration assay) were acquired with a Perkin Elmer Ultraview ERS (Norwalk, CT) disk-spinning confocal system, mounted on a Zeiss Axiovert 200M inverted microscope. To account for the axial focal changes of cells as they move, optical Z-sections were collected at 0.95 μm interval spacing (Movies: Supplementary data M1 & M2 ).
3
Visualizing Mitochondrial Transfer via TnTs
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Time-lapse imaging experiments were performed on Perkin Elmer UltraView ERS spinning-disk confocal microscope or Zeiss LSM 5Live line-scanning confocal microscope. Both microscopes were enclosed in environmental chambers that were maintained at 37°C with 5% CO2 level. Viable Staining of Cell Lines for time-lapse imaging was performed as we have described previously (Lou et al., 2012b (link)). Briefly, in order to assess the ability of mitochondria to be transmitted between mesothelioma cells via TnTs, we used MitoTracker Red to stain MSTO-211H cells which were then cultured in hyperglycemic, low-serum (“TnT”) medium. The cells were cultured in clear-bottomed delta-T culture dishes (Bioptechs Inc., Butler, PA). MitoTracker Red CMX Ros (Invitrogen, M-7512, 50 μg/vial) was used at 500 nM to stain mitochondria, per manufacturer's protocols. Stained cells were re-suspended and added to a non-confluent culture of adherent, unstained MSTO-211H cells grown in another dish. Incubation was performed in high glucose medium for 5 h to stimulate formation of TnTs prior to imaging.
4
Immunofluorescence Microscopy of Cultured Cells
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Cells cultured on glass coverslips (Corning) were washed with PBS and fixed using 4% PFA and permeabilized using 0.25% Triton X-100. For imaging after sorting, cells were suspended in PBS at concentrations of ~ 5 × 106 cells/ml and fixed to coverslips by centrifugation. After blocking in 5% BSA, cells were incubated with primary antibodies at 4 °C, washed with PBS, and incubated with highly cross-adsorbed, AlexaFluor-conjugated secondary antibodies at room temperature. Coverslips were mounted to slides using Vectashield hard-set mounting medium with DAPI (Vector Laboratories). Images were acquired using a PerkinElmer UltraVIEW ERS spinning disc confocal imager mounted on a Zeiss AxioVert 200 inverted microscope equipped with a 63×/1.4 Oil DIC Plan-Apochromat 0.19/0.17. Primary antibodies used for fluorescence microscopy were: TFAM (Cell Signaling), cyclin B1 [GNS1] (SCBT), 8-oxo-dG [15A3] (SCBT), COXIV (Cell Signaling), and MAP LC3β [G-9] (SCBT).
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