Bz x710 series

Manufactured by Keyence

The BZ-X710 series is a line of fluorescence microscopes designed for biological and life science research. It features high-speed image acquisition, advanced imaging capabilities, and a user-friendly interface. The BZ-X710 series enables researchers to capture and analyze fluorescent samples with precision and efficiency.

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Lab products found in correlation

Streptavidin hrp, by Merck Group (1 mentions) Anti phospho ripk3, by Cell Signaling Technology (1 mentions) Anti phospho mlkl, by Cell Signaling Technology (1 mentions) Immunocal, by StatLab (1 mentions) Biotinylated anti gl7 antibodies, by Thermo Fisher Scientific (1 mentions) Texasred conjugated anti mouse igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 568 conjugated streptavidin, by Thermo Fisher Scientific (1 mentions) Bz x viewer, by Keyence (1 mentions) Plan fluor, by Keyence (1 mentions)

3 protocols using bz x710 series

Visualizing Protein Phase Separation

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Purified fluorescence-tagged protein in 450 mM salt buffer was diluted to 125 mM KCl in 10 mM HEPES, pH 7.4, in 96-well plates. The plates were incubated at 37 °C and images were captured at indicated time points after incubation. All phase separation of purified protein was performed in phase-separation buffer (10 mM HEPES, pH 7.4, 125 mM KCl) without adding any crowding reagents. For RIPK3_295–518–mCherry and ZBP1–BFP recruitment to OASL–GFP droplets, variant GFP-tagged OASL proteins, mCherry-tagged RIPK3_295–518 and BFP-tagged ZBP1 were mixed and incubated in 96-well plates at 37 °C for 1 h at the indicated concentrations in the phase-separation buffer. Image acquisition of phase separation was performed using a fluorescence microscope (Keyence, BZ-X710 series) under ×20, ×40 and ×60 1.49-NA oil-immersion objectives. At least four independent imaging areas were analysed for each condition of each replicate. Data shown are representative of at least three independent experiments across five protein preparations.

Lee S.A., Chang L.C., Jung W., Bowman J.W., Kim D., Chen W., Foo S.S., Choi Y.J., Choi U.Y., Bowling A., Yoo J.S, & Jung J.U. (2023). OASL phase condensation induces amyloid-like fibrillation of RIPK3 to promote virus-induced necroptosis. Nature Cell Biology, 25(1), 92-107.

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Quantification of RIPK3 and MLKL Phosphorylation in Mouse Footpads

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Mouse footpads were collected, fixed in 4% paraformaldehyde for 24 h and transferred to 70% ethanol for storage. Tissue sectioning and immunohistochemistry were performed by the University of Southern California Immunohistochemistry Core facility. In brief, footpads were decalcified in formic acid (Immunocal, StatLab), embedded in paraffin and sectioned at 5 μm thickness. Slides were deparaffinized and either stained with haematoxylin and eosin or antigen-retrieved using retrieval buffer for immunohistochemistry staining. Anti-phospho RIPK3 (Cell Signaling Technology) and anti-phospho MLKL (Cell Signaling Technology) were used for staining. Staining was visualized using streptavidin–HRP (Millipore) and DAB substrate (DAKO and Vector Lab). All immunohistochemistry sections were counterstained with haematoxylin. Images were captured using a bright-field microscope (Keyence, BZ-X710 series). Quantification of phosphorylation signals, epidermatitis or inflamed area were calculated into percentage values (positive signal area versus total area of field of view) on individual footpad cross-sections.

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Kidney and Spleen Immunofluorescence Assay

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Half kidneys were frozen in OCT and 5μm sections were prepared and stained using Texas Red-conjugated anti-mouse IgG (Invitrogen, Waltham, MA, USA) and fluorescein isothyocyanate (FITC)–conjugated anti-mouse C3 specific antibodies (Immunology Consultants Laboratory, Portland, OR, USA). Images were obtained using a 10x/0.3 Nikon Plan Fluor objective lens on a Keyence BZ-X710 Series fluorescence microscope. IgG deposition was quantified using Keyence BZ-X Viewer and Keyence BZ-X Analyzer software (Version 1.3.1.1, Keyence Corp., Osaka, Japan) and reported as arbitrary units representing average integrated brightness per glomerulus. Mean brightness of glomeruli per mouse were quantified by averaging integrated brightness of each glomerulus within a 10X field of view. Detection of GC B cells within spleens was done using FITC-conjugated anti-B220 antibodies, biotinylated anti-GL7 antibodies (EBiosciences), and Alexa Fluor 568-conjugated streptavidin (Invitrogen) on 5μm frozen sections. Mean area of GCs per mouse were quantified by averaging area of each GC within 10X field of view using microscope software mentioned above. All images to be compared were obtained using identical microscope settings (gain and exposure time).

Campbell N.O., Davison L.M., Banerjee S., Nguyen J.K., Krafcik S., Silverman R.H, & Jorgensen T.N. (2022). Ablation of SigH+ pDCs in B6.Nba2 mice prevents lupus-like disease development only if started before disease is fully established. Lupus, 31(13), 1619-1629.

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