Orca 2 er camera

Manufactured by Hamamatsu Photonics

Sourced in Japan, France

The ORCA II ER camera is a high-performance scientific imaging device designed for low-light applications. It features a back-illuminated CCD sensor with high quantum efficiency, low readout noise, and high dynamic range. The camera is capable of capturing high-quality images and video with excellent sensitivity and resolution.

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Lab products found in correlation

Ultraview rs nipkow disk, by Zeiss (2 mentions) Axiovert 200m, by Zeiss (2 mentions) Nis element software, by Nikon (1 mentions) Eclipse ti u, by Hamamatsu Photonics (1 mentions) Eclipse 80i microscope, by Hamamatsu Photonics (1 mentions) Orca er camera, by Hamamatsu Photonics (1 mentions) Metamorph software, by Molecular Devices (1 mentions) Syto 61, by Thermo Fisher Scientific (1 mentions) 24 mm glass coverslips, by Marienfeld (1 mentions) Orca flash 4, by Hamamatsu Photonics (1 mentions) Axiovert s100 tv2 microscope, by Zeiss (1 mentions) Leica dmi6000 microscope, by Leica Microsystems (1 mentions) Axiovert 200m, by Hamamatsu Photonics (1 mentions)

6 protocols using orca 2 er camera

Microscopic Observation of Brucella and E. coli

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Brucella strains and infected HeLa cells were observed with a Nikon 80i (objective phase contrast × 100, plan Apo) connected to a Hamamatsu ORCA-ER camera. For the observation of bacteria, 1.5 μl of an exponential phase culture was dropped on agarose pad (solution of 1% agarose in PBS) and sealed with VALAP (1/3 of vaseline, 1/3 of lanoline and 1/3 of paraffin wax).
Time-lapse experiments with B. abortus strains were performed from cultures in exponential phase in liquid Plommet–erythritol containing K₂HPO₄ (0.92 g l⁻¹), KH₂PO₄ (3 g l⁻¹), Na₂S₂O₃ (0.1 g l⁻¹), NaCl (5 g l⁻¹), Nicotinate (0.2 g l⁻¹), Thiamine (0.2 g l⁻¹), (NH₄)₂SO₄ (0.5 g l⁻¹), Pantothenate (0.07 g l⁻¹), Erythritol (2 g l⁻¹), MgSO₄·7H₂O (10 g l⁻¹), MnSO₄ (1.1 g l⁻¹), FeSO₄·7H₂O (1 g l⁻¹) and Biotine (1 g l⁻¹). Cultures were spread on a sealed Plommet–erythritol agarose 1% pad. Images were automatically taken every 20 min at 32 °C with NIS-Element software (Nikon).
Imaging of E. coli strains was performed using either a Nikon Eclipse 80i microscope equipped with a Hamamatsu Orca II-ER camera ( × 100 differential interference contrast or phase-contrast objectives, optivar × 1.5 when appropriate) or a Nikon Eclipse Ti-U with a Hamamatsu Orca-ER LCD camera ( × 100 phase-contrast objective). Images were processed with Metamorph software (Molecular Devices, Sunnyvale, CA).

Deghelt M., Mullier C., Sternon J.F., Francis N., Laloux G., Dotreppe D., Van der Henst C., Jacobs-Wagner C., Letesson J.J, & De Bolle X. (2014). G1-arrested newborn cells are the predominant infectious form of the pathogen Brucella abortus. Nature Communications, 5, 4366.

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3D Confocal Imaging and Quantification

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Confocal acquisitions were performed using a spinning-disk Perkin-Elmer Ultraview RS Nipkow Disk, an inverted laser-scanning confocal microscope Zeiss Axiovert 200M with an Apochromat × 100/1.4 oil objective and a Hamamatsu ORCA II ER camera (Imagopole, PFID, Institut Pasteur). Optical slices were taken at 200-nm intervals along the z axis covering the whole depth of the cell, at a 1.024/1.024 pixel resolution. Three-dimensional reconstruction was achieved using the IMARIS software (Bitplane). Fluorescence quantification was done using a single-imaging frame collection and ImageJ 1.47-v software (post-acquisition analysis).

Rocheteau P., Chatre L., Briand D., Mebarki M., Jouvion G., Bardon J., Crochemore C., Serrani P., Lecci P.P., Latil M., Matot B., Carlier P.G., Latronico N., Huchet C., Lafoux A., Sharshar T., Ricchetti M, & Chrétien F. (2015). Sepsis induces long-term metabolic and mitochondrial muscle stem cell dysfunction amenable by mesenchymal stem cell therapy. Nature Communications, 6, 10145.

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Visualization of Autophagy in MCF-7 Cells

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MCF-7 cells were seeded overnight into 4 well chamber slides at a density of 1.5 × 10⁴ cells/chamber followed by a 72-hours treatment with or without either 10 μM 8-Cl-Ado or 50 nM rapamycin. AVO were stained by incubating cells in 50 mM MDC (Sigma-Aldrich) in PBS at 37°C for 10 minutes and then washed three times with PBS. Nuclei were counter stained with 5 μM SYTO 61 (Invitrogen) in Tris-buffered saline (25 mM Tris, 150 mM NaCl, pH 7.5) at 37°C for 10 minutes and then washed twice with PBS and immediately analyzed at UT MD Anderson Cancer Center’s Flow Cytometry and Cellular Imaging Core Facility by fluorescence microscopy using an inverted microscope (Olympus 1X71, Melville, NY) equipped with a filter system (excitation filter: 350/50 nm, emission filter: 528/38 nm). Images were obtained with a Hamamatsu Orca II ER camera (Hamamatsu, Japan) and processed using the program Slidebook (3I, Denver, CO).

Stellrecht C.M., Vangapandu H.V., Le X.F., Mao W, & Shentu S. (2014). ATP directed agent, 8-chloro-adenosine, induces AMP activated protein kinase activity, leading to autophagic cell death in breast cancer cells. Journal of Hematology & Oncology, 7, 23.

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Imaging Monocyte Dynamics and Death

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Peripheral blood mononuclear cells were plated at 5 × 10⁶/well in 24 mm glass coverslips (Marienfeld, Germany) for 30 min, washed in PBS and incubated in complete medium at 37 °C with different stimuli. Time-lapse photographs were then taken every 3 min for up to 6 h using a Zeiss Axiovert S100 TV2 microscope (Zeiss) equipped with a 63 × /1.4 lens and a Hamamatsu OrcaII-ER camera (Hamamatsu City). Images were analyzed using ImageJ or Oko-vision (Okolab) softwares^{54 (link)}. Monocytes stimulated for 6 h with LPS or LRZ or for 3 h with LPS and then with R848 and zymosan for 3 h were stained for 10 min with PI (5μg/ml) and images were recorded with the software LASX on a Leica DMI6000 microscope (Leica Microsystems, Mannheim, Germany) with a 63 × /1.45 objective and a sCMOS camera (Hamamatsu Flash ORCA 4.0).

Semino C., Carta S., Gattorno M., Sitia R, & Rubartelli A. (2018). Progressive waves of IL-1β release by primary human monocytes via sequential activation of vesicular and gasdermin D-mediated secretory pathways. Cell Death & Disease, 9(11), 1088.

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High-Resolution 3D Confocal Imaging

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Confocal acquisitions were performed using a spinning-disk Perkin-Elmer Ultraview RS Nipkow Disk, an inverted laser-scanning confocal microscope Zeiss Axiovert 200 M with an Apochromat 63X/1.4 NA oil objective and a Hamamatsu ORCA II ER camera (Institut Pasteur, Paris). Microphotographs were acquired using non-saturating settings, and the same imaging parameters were used for all samples. Optical z-slices in 200-nm steps covering the whole depth of the cell were collected, at resolution of 1.024/1.024 pixels. Three-dimensional reconstruction of all the z stacks was achieved using the 3D-volume rendering of IMARIS software (Bitplane). Representative 2D images generated from 3D volume rendering were used for fluorescence quantification using the Integrated Density measurement tool of ImageJ 2.0.0 software (post-acquisition analysis).

Arena G., Cissé M.Y., Pyrdziak S., Chatre L., Riscal R., Fuentes M., Arnold J.J., Kastner M., Gayte L., Bertrand-Gaday C., Nay K., Angebault-Prouteau C., Murray K., Chabi B., Koechlin-Ramonatxo C., Orsetti B., Vincent C., Casas F., Marine J.C., Etienne-Manneville S., Bernex F., Lombés A., Cameron C.E., Dubouchaud H., Ricchetti M., Linares L.K, & Le Cam L. (2018). Mitochondrial MDM2 Regulates Respiratory Complex I Activity Independently of p53. Molecular cell, 69(4), 594-609.e8.

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Measuring Circadian Rhythms in Arabidopsis RILs

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We measured circadian period and phase on eight replicates per RIL for each of the three RIL populations at three time points (June, July, and September) over the course of the growing season, as described in detail in Rubin et al. (2017) (link). In brief, seeds were surface sterilized and pipetted into 96-well white microtiter plates containing Murashige and Skoog media. Following cold stratification of seed, the plates were moved into a Percival PGC-9/2 growth chamber to germinate (12-hour photoperiod, 22°, and 50% relative humidity). After seed germination, seedlings were entrained in the field for 5 days, a period of time sufficient for entrainment to field conditions. Plates were then returned to the lab for bioluminescence imaging. D-luciferin monopotassium salt was added to each well, and the plants were imaged for 4 days using a Hamamatsu ORCA IIER camera (Hamamatsu Photonics C4742-98-24ER). We estimated circadian period and phase using ImagePro/IandA software (Plautz et al. 1997 (link); Mcwatters et al. 2000 (link); Doyle et al. 2002 (link)). Circadian period is defined as the average cycle length, and circadian phase is the timing of peak expression relative to dawn on the final day of field entrainment (Rubin et al. 2017 (link); Rubin et al. 2018 (link)).

Rubin M.J., Brock M.T., Davis S.J, & Weinig C. (2019). QTL Underlying Circadian Clock Parameters Under Seasonally Variable Field Settings in Arabidopsis thaliana. G3: Genes|Genomes|Genetics, 9(4), 1131-1139.

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