Ultraview vox imaging system

Manufactured by PerkinElmer

The UltraVIEW VoX imaging system is a high-performance confocal microscope designed for live-cell imaging. It features a modular design, allowing for customization to meet specific research needs. The system provides high-resolution, real-time imaging capabilities for a wide range of applications.

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Lab products found in correlation

Mitotracker red, by Thermo Fisher Scientific (3 mentions) Phalloidin 488, by Thermo Fisher Scientific (1 mentions) Prolong diamond antifade mountant, by Thermo Fisher Scientific (1 mentions) Recombinant mouse tgf β, by R&D Systems (1 mentions) Recombinant mouse il 1α, by R&D Systems (1 mentions) Anti α sma antibody, by Thermo Fisher Scientific (1 mentions) Influx cell sorter, by BD (1 mentions) Volocity, by PerkinElmer (1 mentions) Lightning link kits, by Novus Biologicals (1 mentions) Csu 1 spinning disk, by Yokogawa (1 mentions) Volocity software, by PerkinElmer (1 mentions) Edu click 488 kit, by Merck Group (1 mentions)

6 protocols using ultraview vox imaging system

Live Cell Imaging and Mitochondrial Fusion Assays

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For live cell imaging, HeLa cells were exposed to hypoxia for 6 h and incubated for 30 min with 50 nM MitoTracker Red (M7512; Thermo Fisher Scientific), washed twice with PBS buffer, and replenished with fresh media. The imaging experiment was performed using a spinning-disk UltraVIEW VoX imaging system (PerkinElmer) equipped with a 60 × 1.4 NA objective lens. For time tracking, cells were observed in a chamber at 37°C under 1% O₂ and images were postprocessed using the Velocity (Nikon) software.
For mitochondrial fusion assays, HeLa cells transfected with mito-PAGFP were imaged using a spinning-disk UltraVIEW VoX microscope system with a 60 × 1.4 NA objective lens. Four-μm-wide ROIs were selected and activated by a single-pulse 405-nm laser. The green fluorescent z-stacks were acquired before and immediately following activation and then every 5 min for 30 min

Chai P., Cheng Y., Hou C., Yin L., Zhang D., Hu Y., Chen Q., Zheng P., Teng J, & Chen J. (2021). USP19 promotes hypoxia-induced mitochondrial division via FUNDC1 at ER-mitochondria contact sites. The Journal of Cell Biology, 220(7), e202010006.

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Hepatic Stellate Cell Cytoskeletal Analysis

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Primary murine hepatic stellate cells were isolated as previously described^{46 (link)}. Following sorting on a BD Influx Cell Sorter in the University of Virginia Flow Cytometry Core, cells were seeded onto fibronectin coated 4 kPa polyacrylamide hydrogels (Matrigen) and stimulated with 10 ng/mL recombinant mouse IL-1α (R&D), 10 ng/mL recombinant mouse TGF-β (R&D), or media alone for 48 h, after which hydrogels were fixed with 4% paraformaldehyde and stained with phalloidin-488 (Invitrogen) and anti α-SMA antibody (Invitrogen, clone IA4). Cells were mounted with ProLong Diamond Anti-Fade mountant (Thermo Fisher Scientific), and imaged at room temperature on a Nikon Eclipse Ti microscope with an UltraView VoX imaging system (PerkinElmer) using a Nikon N Apo LWD 40 × water objective (numerical aperture: 1.15) and cell area and α-SMA intensity were determined using Volocity software.

Melchor S.J., Hatter J.A., Castillo É.A., Saunders C.M., Byrnes K.A., Sanders I., Abebayehu D., Barker T.H, & Ewald S.E. (2020). T. gondii infection induces IL-1R dependent chronic cachexia and perivascular fibrosis in the liver and skeletal muscle. Scientific Reports, 10, 15724.

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Visualizing Intracellular Calcium Dynamics

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HeLa cells stably expressing mitochondria-targeted R-GECO1.2 (mito-R-GECO1.2), which acts as mitochondria Ca²⁺ sensor and emits red fluorescence, or cyto-GCaMP6s, which acts as cytosol Ca²⁺ sensor and emits green fluorescence, were used for the Ca²⁺ imaging experiments. Ca²⁺ imaging was performed with a spinning-disk UltraVIEW VoX imaging system (Perkin Elmer) equipped with a 20 × /0.75 NA objective lens (Nikon).
For histamine-induced Ca²⁺ dynamics, cells were washed twice with Hank’s balanced salt solution, followed by the addition of Ca²⁺ buffer (150 mM NaCl, 5.4 mM KCl, 20 mM HEPES, 10 mM glucose, 1 mM MgSO₄, and 1.8 mM CaCl₂, pH 7.4), immediately before imaging. Cells were excited at either 561 nm (for mitochondria Ca²⁺) or 488 nm (for cytosol Ca²⁺), and images were acquired every 5 s for 5 min. Approximately 30 s after the start of the experiment, histamine was added at a final concentration of 100 μM. Images were post-processed with Volocity (Perkin Elmer).
For mitochondria Ca²⁺ upon DNA damage, cells were treated with 1 μM cpt for 10 h before imaging. Images were acquired at 30 points randomly, and post-processed and analyzed with Volocity (Perkin Elmer).

Zheng P., Chen Q., Tian X., Qian N., Chai P., Liu B., Hu J., Blackstone C., Zhu D., Teng J, & Chen J. (2018). DNA damage triggers tubular endoplasmic reticulum extension to promote apoptosis by facilitating ER-mitochondria signaling. Cell Research, 28(8), 833-854.

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Immunofluorescence Staining of Cells

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Cells were washed twice with PBS and adhered to covers pre‐coated with poly‐l‐lysine, followed by fixation with 4% paraformaldehyde for 10 min at room temperature. Cells were then washed twice with PBS and permeabilized with PBS containing 1% Triton X‐100 for 10 min, followed by washing with PBS for twice and blockage in 10% normal goat serum for 30 min at 37°C. Cells were incubated with primary antibody for 2 h, followed by washing with PBS for three times and further incubation with fluorescence‐conjugated secondary antibody for 1 h. Nuclei were stained with DAPI. For co‐immunostaining using antibodies with the same origin, Lightning‐Link kits (ab236553 and ab269900, Novus Biologicals) were used to directly link fluorescence to primary antibodies. Cells were visualized through an UltraVIEW VoX imaging system (PerkinElmer).

Ma J., Zhu F., Zhao M., Shao F., Yu D., Ma J., Zhang X., Li W., Qian Y., Zhang Y., Jiang D., Wang S, & Xia P. (2021). SARS‐CoV‐2 nucleocapsid suppresses host pyroptosis by blocking Gasdermin D cleavage. The EMBO Journal, 40(18), e108249.

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Immunofluorescence Staining of Confluent HUVECs

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Confluent HUVECs were washed with PBS and fixed in 4% paraformaldehyde for 10 min at 37°C. Cells were then washed three times and permeabilized with 0.01% Triton X-100 for 10 min at room temperature. Blocking buffer (PBS + 2.5% BSA, 2.5% FBS, and 2.5% host species serum) was added for 1 h at room temperature. Cells were then incubated with fluorescently labeled primary Ab at 20 µg/ml in blocking buffer for 1 h at room temperature. Confocal images were visualized using an UltraVIEW VoX imaging system (PerkinElmer) equipped with a CSU-1 spinning disk (Yokogawa Electric Corporation). Images were acquired with a 40× oil immersion objective using Volocity software (PerkinElmer). All images were processed and analyzed with ImageJ software (National Institutes of Health).

Weber E.W., Han F., Tauseef M., Birnbaumer L., Mehta D, & Muller W.A. (2015). TRPC6 is the endothelial calcium channel that regulates leukocyte transendothelial migration during the inflammatory response. The Journal of Experimental Medicine, 212(11), 1883-1899.

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Inflammasome Activation and Mitochondrial DNA Imaging

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Cells adhered to covers pre-coated with poly-L-lysine were stimulated with inflammasome activators, followed by fixation with 4% paraformaldehyde for 10 min at room temperature. Cells were then washed twice with PBS and permeabilized with PBS containing 1% Triton-X-100 for 10 min, followed by washing with PBS for twice and blockage in 10% normal goat serum at 37 C for 30 min. Cells were incubated with primary antibody for 2 h, followed by washing with PBS for three times and further incubation with fluorescenceconjugated secondary antibody for 1 h. Nuclei were stained with DAPI. For EdU staining, WT BMDM cells were stimulated with 200 ng/ml LPS and 10 mM EdU for 3 h to specifically label the mitochondrial DNA as previously reported. 35 (link) A EdU-Click 488 kit (Sigma-Aldrich) was used to visualize EdU inside cells. Cells were visualized through an UltraVIEW VoX imaging system (PerkinElmer).

Zhu F., Ma J., Li W., Liu Q., Qin X., Qian Y., Wang C., Zhang Y., Li Y., Jiang D., Wang S, & Xia P. (2023). The orphan receptor Nur77 binds cytoplasmic LPS to activate the non-canonical NLRP3 inflammasome. Immunity, 56(4).

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