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Orca flash4.0 lt plus scmos camera

Manufactured by Hamamatsu Photonics
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Sourced in Ireland

The Orca-Flash4.0 LT PLUS/sCMOS camera is a high-performance scientific imaging device developed by Hamamatsu Photonics. It features a scientific CMOS (sCMOS) sensor that provides fast readout speeds, high quantum efficiency, and low noise levels. The camera is designed for a variety of scientific and industrial applications that require high-quality image capture and analysis.

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Lab products found in correlation

Spectrax led light source, by Lumencor (2 mentions) Axioimager z2 epifluorescence microscope, by Zeiss (1 mentions) Cy3 conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Eclipse ti2 e n sim e fluorescence microscope, by Nikon (1 mentions) Hydromount, by National Diagnostics (1 mentions) Alexa fluor 488 cy3, by Jackson ImmunoResearch (1 mentions) Abn1376, by Merck Group (1 mentions)

4 protocols using orca flash4.0 lt plus scmos camera

1

High-Resolution Multi-Channel Fluorescence Imaging

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Imaging was performed using a Zeiss Axio Imager.Z2 epifluorescence microscope (Carl Zeiss Microscopy, GmbH), with a Zeiss Plan-Apochromat 20×/0.8 objective (Carl Zeiss Microscopy, GmbH, 420650-9901) and an automatic multi-slide stage (PILine, M-686K011) to allow re-call of coordinates for the regions of interest, facilitating repetitive cycle imaging. The system was equipped with a Lumencor SPECTRA X light engine LED source (Lumencor), having the 395/25, 438/29, 470/24, 555/28, 635/22 and 730/40 filter paddles. The filters, for wavelength separation, included the quad band Chroma 89402 (DAPI, Cy3, Cy5), the quad band Chroma 89403 (AlexaFluor750) and the single band Zeiss 38HE (AlexaFluor488). Images were obtained with an ORCA-Flash4.0 LT Plus sCMOS camera (2,048 × 2,048, 16-bit, Hamamatsu Photonics K. K.).
Sountoulidis A., Marco Salas S., Braun E., Avenel C., Bergenstråhle J., Theelke J., Vicari M., Czarnewski P., Liontos A., Abalo X., Andrusivová Ž., Mirzazadeh R., Asp M., Li X., Hu L., Sariyar S., Martinez Casals A., Ayoglu B., Firsova A., Michaëlsson J., Lundberg E., Wählby C., Sundström E., Linnarsson S., Lundeberg J., Nilsson M, & Samakovlis C. (2023). A topographic atlas defines developmental origins of cell heterogeneity in the human embryonic lung. Nature Cell Biology, 25(2), 351-365.
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2

Retinal Immunocytochemistry and Imaging

Cited 2 times
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Mice were sacrificed, eyes enucleated and fixed overnight in 4% paraformaldehyde in PBS before the retinas were removed from the eyecups and immunocytochemistry performed as described previously described [58 (link)]. Whole retinas were incubated with anti-BRN3A primary antibody (Synaptic Systems 411003, Goettingen, Germany; 1/200 dilution) [59 (link),60 (link)] for 3 days at 4 °C, washed in PBS and then incubated with Cy3 conjugated secondary antibodies (Jackson ImmunoResearch Laboratories; 1/400) for 2 days at 4 °C. Wholemount and cell images were taken using an Olympus IX83 inverted motorised microscope (Mason Technology, Dublin, Ireland) equipped with a SpectraX LED light source (Lumencor, Mason Technology, Dublin, Ireland) and an Orca-Flash4.0 LT PLUS/sCMOS camera (Hamamatsu, Tsukuba City, Japan), as previously described [47 (link)]. Samples were imaged using a 4x (wholemount) and 40× (cells) objective utilising enhanced focal imaging (EFI) with typically 5–8 Z-slices. Lateral frames for wholemounts were stitched together and analysed in Olympus CellSens software (v1.9; Waltham, MA, USA). Cell counting was performed utilizing 2D deconvolution, manual threshold and object size filter, with the same settings/operations applied to all images.
Chadderton N., Palfi A., Maloney D.M., Carrigan M., Finnegan L.K., Hanlon K.S., Shortall C., O’Reilly M., Humphries P., Cassidy L., Kenna P.F., Millington-Ward S, & Farrar G.J. (2023). Optimisation of AAV-NDI1 Significantly Enhances Its Therapeutic Value for Correcting Retinal Mitochondrial Dysfunction. Pharmaceutics, 15(2), 322.
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3

Structured Illumination Fluorescence Microscopy of EGFP Fusion Proteins

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For localization studies of the EGFP fusion proteins, Structured Illumination Fluorescent microscopy (SIM) was performed on an Eclipse Ti2-E N-SIM E fluorescence microscope (Nikon) equipped with a CFI SR Apo TIRF AC 100× H NA1.49 Oil objective lens, a hardware based “perfect focus system” and an Orca Flash4.0 LT Plus sCMOS camera (Hamamatsu). Sample preparation, fluorescence excitation with 488 for imaging GFP and image reconstruction and analysis were performed as reported previously (92 (link)).
Awal R.P., Lefevre C.T, & Schüler D. (2023). Functional expression of foreign magnetosome genes in the alphaproteobacterium Magnetospirillum gryphiswaldense. mBio, 14(4), e03282-22.
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4

Retinal Ganglion Cell Immunostaining

Cited 2 times
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Mice were sacrificed 3 days after OKR assessment and eyes were enucleated and fixed in 4% paraformaldehyde in PBS overnight. Eyes were washed in PBS, then the retinas were removed from the eyecups and immediately processed for immunocytochemistry. Immunocytochemistry was performed as described previously (Palfi et al., 2016 (link)). Whole retinas were incubated with primary antibodies for RBPMS (ABN1376, Millipore, 1:200; Rodriguez et al., 2014 (link)) overnight for 3 days at 4°C. Retinas were then washed in PBS and incubated with secondary antibodies conjugated with Alexa-Fluor-488, Cy3 (Jackson ImmunoResearch Laboratories; 1:400) for 2 days and nuclei counterstained with DAPI. Samples were covered using Hydromount (National Diagnostics). Fluorescent microscopy was carried out utilizing an Olympus IX83 inverted motorized microscope (cellSens v1.9 software) equipped with a SpectraX LED light source (Lumencor) and an Orca-Flash4.0 LT PLUS/sCMOS camera (Hamamatsu). Samples were imaged using a 10x plan fluorite objective utilizing enhanced focal imaging (EFI) with typically 5–8 Z-slices. Lateral frames were stitched together and analyzed in cellSens. Automated cell staining area was calculated utilizing 2D deconvolution, manual threshold and object size filter in cellSense; the same settings/operations were applied to all images.
Maloney D.M., Chadderton N., Millington-Ward S., Palfi A., Shortall C., O’Byrne J.J., Cassidy L., Keegan D., Humphries P., Kenna P, & Farrar G.J. (2020). Optimized OPA1 Isoforms 1 and 7 Provide Therapeutic Benefit in Models of Mitochondrial Dysfunction. Frontiers in Neuroscience, 14, 571479.
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