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Orca flash4.0 lt digital cmos camera

Manufactured by Hamamatsu Photonics
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Sourced in Germany, Japan

The ORCA-Flash4.0 LT Digital CMOS camera is a high-performance imaging device designed for scientific and industrial applications. It features a 4.2-megapixel CMOS sensor with a pixel size of 6.5 μm, and is capable of capturing images at a maximum frame rate of 100 frames per second. The camera offers a dynamic range of 12-bit and provides a low readout noise, making it suitable for a variety of imaging tasks that require high-quality, low-noise data.

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Lab products found in correlation

Axio observer z1, by Zeiss (3 mentions) Axioobserver z1 widefield microscope, by Zeiss (2 mentions) Stereo investigator 10, by MicroBrightField (2 mentions) Axioplan 2 microscope, by Zeiss (2 mentions) Zen 3, by Zeiss (2 mentions) Metamorph 7.5 imaging software, by Molecular Devices (2 mentions) Axio imager, by Zeiss (2 mentions) Planapo 60, by Olympus (1 mentions) Lsm 880 with airyscan, by Zeiss (1 mentions) Zen 2, by Zeiss (1 mentions) Nb300 605, by Novus Biologicals (1 mentions) Triton x 100, by Merck Group (1 mentions) Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions) Metamorph, by Molecular Devices (1 mentions) Ix70 microscope, by Olympus (1 mentions) Axio imager m2 imaging microscope system, by Zeiss (1 mentions) Er tracker, by Thermo Fisher Scientific (1 mentions) Z2 microscope, by Zeiss (1 mentions) Orca er ccd camera, by Hamamatsu Photonics (1 mentions) Zyla 4.2 scmos camera, by Oxford Instruments (1 mentions)

15 protocols using orca flash4.0 lt digital cmos camera

1

Fluorescent Imaging of Microchamber Cell Arrays

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The assembled microchamber system was imaged using a Zeiss Axio Observer.Z1 fluorescent microscope with a Hamamatsu Orca-Flash4.0 LT Digital CMOS camera (C11440-42 U) and an automatic stage. For each microchamber array, bright field images were taken to visualize the microchambers and fluorescent images were taken to visualize cells labeled with CD4 or CD8 antibodies. A set of 253 tiles (5X image) or 494 tiles (10X image) was taken for the entire array (54183uM x 24866uM). Using Zen2 Pro software, the tiles were exported as TIFF image files for analysis.
Xue Q., Bettini E., Paczkowski P., Ng C., Kaiser A., McConnell T., Kodrasi O., Quigley M.F., Heath J., Fan R., Mackay S., Dudley M.E., Kassim S.H, & Zhou J. (2017). Single-cell multiplexed cytokine profiling of CD19 CAR-T cells reveals a diverse landscape of polyfunctional antigen-specific response. Journal for Immunotherapy of Cancer, 5, 85.
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2

Multimodal Imaging of Membrane Infoldings

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Wide-field fluorescence imaging was performed on inverted microscopes (IX83, Olympus) using an oil-immersion PlanApo 60× NA 1.42 objective lens. Digital still images were captured by ORCA-Flash4.0 LT+ digital CMOS camera (Hamamatsu) through the software MicroManager (Open Imaging) (62 (link)). For imaging of membrane infoldings, multiple z-stack images were captured on a confocal microscope (LSM 880 with Airyscan, Carl Zeiss) using an oil-immersion 63× NA 1.4 DIC objective lens. Images were captured with 32-channel GaAsp photomultiplier modules using ZEN 2.3 software (Carl Zeiss). All confocal z-stack images were subjected to Airyscan processing. For imaging and tracking of AChR vesicles, high-speed time-lapse images were captured at 200 ms per frame on live-SR super-resolution module on an iLas3 Ring-TIRF system (Gataca Systems). All acquisition settings (i.e., laser power and gain) were kept the same for different experimental groups in the same experiment. All acquired images were analyzed by ImageJ (NIH) or Imaris (Bitplane).
Kwan H.L., Chan Z.C., Bi X., Kutkowska J., Prószyński T.J., Chan C.B, & Lee C.W. (2023). Nerve-independent formation of membrane infoldings at topologically complex postsynaptic apparatus by caveolin-3. Science Advances, 9(24), eadg0183.
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3

Staining of M1 Macrophages on Chip

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We fixed and stained the cells cultured on the chip after day 15. Cells were fixed with 4% paraformaldehyde (15711, Electron Microscopy Sciences) for 30 min, permeabilized with 0.3% Triton X- 100 (11332481001, Sigma-Aldrich) for 10 min, and then blocked with 3% bovine serum for 1 hr on ice to eliminate nonspecific binding. Specifically, we stained M1 phenotype macrophages by incubating the chip with iNOS primary antibodies (NB300–605, Novus Biologicals, 5 μg/mL) for 1 hr, and then visualized with Alexa Fluor 555 conjugated goat anti-mouse IgG secondary antibodies (Invitrogen, 5 mg/mL). All cell nucleus was stained with DAPI. Fluorescent images were obtained using an inverted microscope (Zeiss Axio Observer.Z1) equipped with a digital CMOS camera (ORCA-Flash4.0 LT Digital CMOS camera, Hamamatsu Photonics) and a 10x objective.
Zhu J., He J., Verano M., Brimmo A.T., Glia A., Qasaimeh M.A., Chen P., Aleman J.O, & Chen W. (2018). An Integrated Adipose-Tissue-On-Chip Nanoplasmonic Biosensing Platform for Investigating Obesity-associated Inflammation. Lab on a chip, 18(23), 3550-3560.
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4

Microscopic Imaging of Heated Protein

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DIC was conducted on a Zeiss AxioObserver Z1
widefield microscope (Carl Zeiss Inc., Berlin, Germany), with
an ORCA-Flash4.0 LT+ Digital CMOS camera (Hamamatsu Photonics,
Hamamatsu, Japan). Images were analyzed using MetaMorph imaging
software (Molecular Devices, CA). Protein solution in PBS was heated
to 60 °C and applied onto a glass slide
(10 μL), shielded with a coverslip, and imaged immediately.
Hossain M.S., Ji J., Lynch C.J., Guzman M., Nangia S, & Mozhdehi D. (2021). Adaptive Recombinant Nanoworms from Genetically Encodable Star Amphiphiles. Biomacromolecules, 23(3), 863-876.
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5

Live and Apoptotic Cell Imaging Protocol

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We used calcein-AM and propidium iodide as markers for live and apoptotic cells, respectively. Immediately following isolation, we incubated cells in the dark with 1 μM of calcein-AM for 30 min at 37°C. Then, the cells were centrifuged at 750 rpm for 5 min to remove residual extracellular dye, washed, recentrifuged, resuspended in PBS, and left for 10 minutes to ensure optimal retention of the calcein-AM. This was followed by a 5 min incubation with 1.25 μg/ml propidium, after which the cells were pelleted, washed and centrifuged, resuspended in a PBS, and plated on coverslips for imaging using an Olympus IX70 microscope and a 20X objective. Calcein-AM was detected using 488 nm excitation and 510 nm emission wavelengths, and propidium iodide was detected using and 596 nm excitation and 615 nm emission wavelengths. Images (2048 v 2048 pixels) from 4–5 fields per coverslip were captured and analyzed at 20X magnification using an Orca-Flash 4.0 LT digital CMOS camera (Hamamatsu Photonics, Bridgewater, NJ) and μManager 2.0 software (42 ).
Tewari M., Khan M., Verma M., Coppens J., Kemp J.M., Bucholz R., Mercier P, & Egan T.M. (2021). Physiology of cultured human microglia maintained in a defined culture medium. ImmunoHorizons, 5(4), 257-272.
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6

Quantitative Microscopy for Cerebral Cortex

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A Zeiss Axio Imager M2 imaging microscope system (Carl Zeiss, Inc., Jena, Germany) and ORCA-Flash4.0 LT+ Digital CMOS camera (Hamamatsu, Hamamatsu, Japan) connected with computer software Stereo Investigator 10.0 (MicroBrightField, Inc., Williston, VT, USA) was used for image acquisition for the purpose of quantification. Twelve fields within each cerebral cortical sample from each slide were first randomly selected based upon the DAPI channel. The randomly selected RGB images of the cerebral cortex were acquired with cy3 (Red), FITC (Green), and DAPI (Blue) filters, using a 40× objective from the Zeiss Axio Imager M2 imaging microscope system (Figure 2) and ORCA-Flash4.0 LT+ Digital CMOS camera (Figure 3, Figure 4, Figure 5, Figure 6, Figure 7 and Figure 8). Each wavelength was acquired separately using the same camera settings for each channel and then pseudo-merged into an RGB image for analysis in Adobe Photoshop. The optical conditions were rigorously maintained the same for all the analysis sessions.
Hatayama K., Kim B., Chen X., Lim Y.P., Davidson J.O., Bennet L., Gunn A.J, & Stonestreet B.S. (2021). Changes in Cellular Localization of Inter-Alpha Inhibitor Proteins after Cerebral Ischemia in the Near-Term Ovine Fetus. International Journal of Molecular Sciences, 22(19), 10751.
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7

Visualizing Dnj1-GFP Localization in Yeast

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A strain expressing a C-terminal fusion of GFP to Dnj1 was constructed in the background of the dnj1Δ mutant (Supplementary Table S1). The Dnj1-GFP expressing cells were grown overnight in YNB+0.5% glucose and stained for 30min at room temperature with 200nM ER-Tracker™ (Invitrogen, Carlsbad, CA) in Hank’s balanced salt solution with calcium and magnesium or 5μg/ml DAPI in phosphate buffered saline (PBS). Cells were imaged using a Zeiss Plan-Apochromat 100x/1.46 oil lens on a Zeiss Axioplan 2 microscope. Images were obtained using an ORCA-Flash4.0 LT digital CMOS camera (Hamamatsu, Hamamatsu City, Japan). All fluorescent images were processed using Zen 3.0 software (Zeiss, Oberkochen, Germany).
Horianopoulos L.C., Lee C.W., Hu G., Caza M, & Kronstad J.W. (2021). Dnj1 Promotes Virulence in Cryptococcus neoformans by Maintaining Robust Endoplasmic Reticulum Homeostasis Under Temperature Stress. Frontiers in Microbiology, 12, 727039.
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8

Microscopic Visualization of Anaerobic Bacterium

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B. porci DSM 105750T was grown at 37 °C for 4 days in BHI broth supplemented with 10% rumen fluid (Bar Diamond Inc., ID, USA), cysteine (0.05% w/v), and DTT (0.02%) under anaerobic conditions (89.3% N2, 6% CO2, 4.7% H2). Cultures (1 ml) were centrifuged (2 min, 5000 × g) and washed once with PBS. Harvested cells were incubated for 10 min in the dark at room temperature with the membrane stain FM4-64 (1 µg/ml) and the DNA stain 4′,6-diamidino-2-phenylindole (DAPI) (2 µg/ml). After final PBS wash, cells were placed onto microscope slides and visualized by phase contrast and fluorescence microscopy using a Zeiss Axio Imager.Z2 microscope (Zeiss, Jena, Germany) equipped with a Plan-Apochromat x 63 phase contrast objective lens, appropriate filter sets, and an ORCA-Flash 4.0 LT digital CMOS camera (Hamamatsu Photonics, Shizuoka, Japan) using the Zeiss Zen 2 (Blue Edition) software. Brightness and contrast level of the images were adjusted using ImageJ (version 1.50g).
Wylensek D., Hitch T.C., Riedel T., Afrizal A., Kumar N., Wortmann E., Liu T., Devendran S., Lesker T.R., Hernández S.B., Heine V., Buhl E.M., M. D’Agostino P., Cumbo F., Fischöder T., Wyschkon M., Looft T., Parreira V.R., Abt B., Doden H.L., Ly L., Alves J.M., Reichlin M., Flisikowski K., Suarez L.N., Neumann A.P., Suen G., de Wouters T., Rohn S., Lagkouvardos I., Allen-Vercoe E., Spröer C., Bunk B., Taverne-Thiele A.J., Giesbers M., Wells J.M., Neuhaus K., Schnieke A., Cava F., Segata N., Elling L., Strowig T., Ridlon J.M., Gulder T.A., Overmann J, & Clavel T. (2020). A collection of bacterial isolates from the pig intestine reveals functional and taxonomic diversity. Nature Communications, 11, 6389.
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9

Quantitative Imaging of Membrane Contacts

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Most imaging was conducted on a IX71 microscope (Olympus) with a 63x objective (and 1.6x zoom) and an ORCA-Flash4.0 LT Digital CMOS camera (Hamamatsu). Rate constant of FKBP recruitment by sFRB1 was imaged at 10 s intervals with a spinning disc confocal, inverted Axiovert 200 (Zeiss) with a 40x objective and an Orca ER CCD camera (Hamamatsu). Both microscopes were driven by Metamorph 7.5 imaging software (Molecular Devices). Four-color imaging in Fig. 4 was conducted with an Eclipse Ti microscope (Nikon) with a 60x objective (and 1.5x zoom) and a Zyla 4.2 sCMOS camera (Andor), driven by NIS Elements software (Nikon). Unless otherwise indicated imaging was done at 1-3 min intervals for 12-30 mins, at times with between 3-5 0.5 um-spaced z positions. Images analyzed and shown are from a single plane, and not maximum intensity projections. Microscopy experiments applying CIT to membrane contact sites were conducted at 37°C, 5% CO2 and humidity with a stage top incubation system (Tokai Hit).
Wu H.D., Kikuchi M., Dagliyan O., Aragaki A.K., Nakamura H., Dokholyan N.V., Umehara T, & Inoue T. (2020). Rational design and implementation of a chemically inducible hetero-trimerization system. Nature methods, 17(9), 928-936.
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10

Visualizing Protein Coacervates by DIC Microscopy

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Protein coacervates were imaged with DIC using a Zeiss AxioObserver
Z1 widefield microscope (Carl Zeiss Inc., Berlin, Germany) connected
to an ORCA-Flash4.0 LT+ Digital CMOS camera (Hamamatsu Photonics,
Hamamatsu, Japan). After incubation at 308 K for 10 min, the protein
solutions (50 μM in PBS) were deposited onto a glass slide which
was shielded with a coverslip. Images were processed and analyzed
using MetaMorph software (Molecular Devices, version 7.8.8.0) and
ImageJ (NIH, version 1.53f51).
Ji J., Hossain M.S., Krueger E.N., Zhang Z., Nangia S., Carpentier B., Martel M., Nangia S, & Mozhdehi D. (2023). Lipidation Alters the Structure and Hydration of Myristoylated Intrinsically Disordered Proteins. Biomacromolecules, 24(3), 1244-1257.
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