N storm a1

Manufactured by Nikon

Sourced in Japan, United States

The N-STORM & A1 is a high-performance microscopy system designed for super-resolution imaging. It utilizes advanced techniques like Stochastic Optical Reconstruction Microscopy (STORM) to achieve resolutions beyond the diffraction limit of light. The system combines a high-sensitivity camera, precise optical components, and specialized software to enable detailed visualization and analysis of cellular structures and dynamics.

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Lab products found in correlation

Antifade mounting medium with dapi, by Beyotime (3 mentions) Fv1000, by Olympus (3 mentions) Alexa fluor 594conjugated goat anti mouse igg h l, by Thermo Fisher Scientific (2 mentions) Primary antibodies for tnf α, by Proteintech (2 mentions) Alexa fluor 488 conjugated goat anti rabbit igg h l, by Thermo Fisher Scientific (2 mentions) Phalloidin, by Yeasen (2 mentions) Phalloidin, by Nikon (2 mentions) Pkh26, by Merck Group (2 mentions) Laser scanning confocal microscope, by Nikon (2 mentions) Bafilomycin a1, by GLPBIO (2 mentions) Cytochalasin d, by GLPBIO (2 mentions) Wortmannin, by GLPBIO (2 mentions) Pkh26, by GLPBIO (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Fluo 4 am, by Thermo Fisher Scientific (2 mentions) Image pro plus, by Nikon (1 mentions) Evos xl core, by Thermo Fisher Scientific (1 mentions) A1r mp, by Nikon (1 mentions) Duolink in situ detection reagent, by Merck Group (1 mentions) Goat anti mouse igg fitc, by Jackson ImmunoResearch (1 mentions)

16 protocols using n storm a1

Immunofluorescence Staining of Embryos

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The embryo was xed with 4% PFA for 15 min at room temperature and 0.3% Triton X-100 in PBS was added for 20 min to permeabilize the cell membrane. Then, the embryo was incubated with primary antibodies for TNF-α (1:100, Proteintech Inc., Rosemont, IL, USA) and CDX2 (1:100, Abcam, Inc., Cambridge, MA, USA) at 4 °C overnight. Subsequently, the embryo was incubated with Alexa Fluor 594conjugated goat anti-mouse IgG (H + L), (1:1000, Invitrogen, Inc., Carlsbad, CA, USA) and AlexaFluor 488conjugated goat anti-rabbit IgG (H + L) (1:1000, Invitrogen, Inc.) for 60 min at room temperature in the dark, followed by mounting with antifade mounting medium with DAPI (Beyotime, Inc., Shanghai, China).
The analysis was conducted using a confocal laser scanning microscope (Olympus FV1000, Olympus Corporation, Waltham, MA, USA) (Nikon N-STORM & A1, Nikon Corporation, Tokyo, Japan). The six embryos used for staining originated from the same volunteer.

Lv J., Shan X., Yang H., Wen Y., Zhang X., Chen H., Li H., Tian D., Wang C.C., Zhang R., Li T.C., Zhang X., Zhao X., Lu Y., Qin L., Zhu M, & Xu W. (2022). Single Cell Proteomics Profiling Reveals That Embryo-Secreted TNF-α Plays a Critical Role During Embryo Implantation to the Endometrium. Reproductive sciences (Thousand Oaks, Calif.), 29(5).

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Embryonic Protein Localization via IF

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The embryo was xed with 4% PFA for 15 min at room temperature and add 0.3% Triton X-100 in PBS for 20 min to change cell membrane permeability. Then the embryo was incubated with TNF-α primary antibody (1:100, Proteintech, Inc.) and CDX2 primary antibody (1:100, Abcam, Inc), at 4℃ overnight. Subsequently, the embryo was incubated with alexa uor 594-conjugated goat anti-mouse IgG (H+L),
(1:1000, Invitrogen, Inc.) and 488-conjugated goat anti-rabbit IgG (H+L), (1:1000, Invitrogen, Inc.), for 60 min at room temperature in dark place, followed by mounting with antifade mounting medium with DAPI (Beyotime, Inc., Shanghai, China). Then, analysis was conducted using a confocal laser scanning microscope (Olympus FV1000, Olympus Corporation) (Nikon N-STORM & A1, Nikon Corporation). The six embryos used for staining came from the same volunteer.

Lv J., Shan X., Yang H., Wen Y., Xue-guang Z., Chen H., Li H., Dongmei T., Wang C.C., Zhang R., Chiu L.T., Xiao-hu Z., Xiaomiao Z., Lu Y., Qin L., Zhu M., & Wenming X. (2020). Single Cell Proteomics Profiling Revealing that Embryo Secreted TNF-α play Critical Role during Embryo Implantation to Endometrium.

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Immunofluorescence Staining of Embryos

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Lv J., Shan X., Yang H., Wen Y., Zhang X., Chen H., Li H., Tian D., Wang C.C., Zhang R., Li T.C., Zhang X., Zhao X., Lü Y., Qin L., Zhu M., & Xu W. (2020). Single cell proteomics profiling reveals that embryo-secreted TNF-α plays a critical role during embryo implantation to the endometrium.

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Immunofluorescence Analysis of Renal Proteins

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Cells and frozen renal tissue sections were fixed with 4% paraformaldehyde in PBS for 10 min at room temperature, washed with PBS, and permeabilized with 0.3% Triton X-100 for 10 min. After blocking in 1% BSA for 30 min, the cells were immunolabeled with primary antibodies (anti-Kim-1 at 1:150 dilution; anti-TNF-α at 1:100 dilution; anti-dsDNA at 1:400 dilution; anti-TOM20 at 1:100 dilution; or anti-TFAM at 1:50 dilution) overnight at 4 °C followed by incubation with FITC- or TRITC-conjugated secondary antibody (1:200) for 1 h at 37 °C. Nuclei were visualized by staining with DAPI for 5 min at room temperature. Digital images of the sections were captured using a fluorescence microscope (Nikon, N-STORM & A1, Tokyo, Japan), and the Pearson's correlation coefficient between the TFAM and dsDNA results was analyzed using Image-Pro Plus.

Zhao M., Wang Y., Li L., Liu S., Wang C., Yuan Y., Yang G., Chen Y., Cheng J., Lu Y, & Liu J. (2021). Mitochondrial ROS promote mitochondrial dysfunction and inflammation in ischemic acute kidney injury by disrupting TFAM-mediated mtDNA maintenance. Theranostics, 11(4), 1845-1863.

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Immunofluorescence Analysis of Kidney Injury

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Human renal tissues that had sustained renal injury were obtained from renal biopsies of AKI patients. Normal kidney tissues obtained from surgical nephrectomy of renal carcinoma patients were used as controls. The process of human sample collection was performed in accordance with the guidelines established by the hospital ethics committee, and informed consent was obtained from the patients according to the Declaration of Helsinki. For IF staining, frozen renal sections were stained with primary antibodies (anti-TOM20, 1:100; anti-NGAL, 1:100; anti-dsDNA, 1:400; anti-TFAM, 1:50) overnight at 4 °C followed by incubation with an FITC- or TRITC-conjugated secondary antibody (1:200) for 1 h at 37 °C. Nuclei were visualized by staining with DAPI for 5 min at room temperature. Digital images were captured using a fluorescence microscope (Nikon, N-STORM & A1, Tokyo, Japan).

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Micropatterned Culture of HepG2 Cellular Aggregates

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HepG2
cells (2 × 10⁵) were seeded on DLM-patterned dishes
containing 3 mL of MEM media to have an equal cell density on the
micropatterned dishes. The culture media was sucked out after 6 h,
and the dishes were washed thrice with PBS to remove the unattached
cells. The cellular aggregates’ morphology at 6 h and on days
1, 2, 3, 4, 5, and 6 after seeding was observed and imaged using EVOS
XL Core (Invitrogen), and their diameters were analyzed using the
ImageJ software (National Institutes of Health, Bethesda, MD). Their
viability was assessed through FluoroQuench fluorescent staining followed
by imaging using a confocal microscope (N-STORM & A1, Nikon, Japan).
Cellular aggregates grown on micropatterned dishes were fixed with
4% formaldehyde for 20 min at room temperature, stained with rhodamine
phalloidin solution (100 nM) (Cytoskeleton) for 45 min at room temperature,
and counterstained with DAPI. Fluorescent images were acquired with
a two-photon confocal microscope (A1RMP+, Nikon, Japan).

Zhu X., Wu Q., He Y., Gao M., Li Y., Peng W., Li S., Liu Y., Zhang R, & Bao J. (2022). Fabrication of Size-Controllable and Arrangement-Orderly HepG2 Spheroids for Drug Screening via Decellularized Liver Matrix-Derived Micropattern Array Chips. ACS Omega, 7(2), 2364-2376.

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In Situ CD44-FGFR2 Protein Interaction

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PLA was used to examine CD44-FGFR2 protein interactions in situ using the Duolink^® In Situ Detection Reagents (Sigma, DUO92014-100RXN). HUVECs were plated on fibronectin-coated glass slides in a 48-well plate. In brief, then, slides were fixed and permeabilized, and incubated with blocking solution for 30 min at 37 °C, primary antibody CD44 Rabbit antibody (Proteintech, 15675-1-AP, 1:200), FGFR2 Mouse antibody (Abcam, ab58201, 1:50), LaminA/C Mouse antibody (CST, 4777, 1:100) and CD9 Mouse antibody (Proteintech, 60232-1-Ig, 1:200) overnight at 4 °C, PLA probe solutions for 60 min at 37 °C, Ligation-Ligase solution for 30 min at 37 °C, and Amplification-Polymerase solution for 100 min at 37 °C, sequentially. All incubations were performed without coverslips in a preheated humidity chamber. Images were obtained using confocal microscope (Nikon, N-STORM & A1) with a FITC filter for the detection of PLA signals, and analyzed by ImageJ software.

Zhang Q., Chen L., Huang L., Cheng H., Wang L., Xu L., Hu D., He C., Fu C, & Wei Q. (2022). CD44 promotes angiogenesis in myocardial infarction through regulating plasma exosome uptake and further enhancing FGFR2 signaling transduction. Molecular Medicine, 28, 145.

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Immunofluorescence Analysis of Kidney Sections

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Frozen sections of kidneys from patients or mice were fixed with 10% formalin and incubated with rabbit anti‐C5aR (Abcam), rabbit anti‐C5a (Abcam), rabbit anti‐p‐signal transducer and activator of transcription (STAT) 3 (Abclonal), mouse anti‐p‐STAT3 (Santa Cruz) and mouse anti‐CD31 (Santa Cruz) antibodies at 4°C overnight. After washing with PBS, the sections were incubated with diluted fluorescence‐conjugated secondary antibodies, including goat anti‐rabbit IgG/TRITC (Jackson ImmunoResearch) and goat antimouse IgG/FITC (Jackson ImmunoResearch), at 37°C in the dark for 1 hour and then stained with DAPI (Calbiochem). Micrographs of the stained sections were acquired on a confocal laser scanning microscope (N‐STORM & A1, Nikon Tokyo). Morphologic analysis of the images was performed with ImageJ software (NIH).

Li L., Wei T., Liu S., Wang C., Zhao M., Feng Y., Ma L., Lu Y., Fu P, & Liu J. (2020). Complement C5 activation promotes type 2 diabetic kidney disease via activating STAT3 pathway and disrupting the gut‐kidney axis. Journal of Cellular and Molecular Medicine, 25(2), 960-974.

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Immunofluorescence Analysis of BMMSCs

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BMMSCs were washed with PBS three times and fixed in 4% paraformaldehyde for 30 min at room temperature. Then cells were blocked in 10% goat serum for 30 min at 37°C and incubated primary antibodies overnight at 4°C. Primary antibodies were diluted as previous description. The next day, cells were placed for 30 min at room temperature and washed with PBS three times. Next, cells were incubated with corresponding secondary fluorescent antibodies rhodamine (TRITC)‐conjugated AffiniPure goat anti‐rabbit IgG (H+L) and/or CoraLite488‐conjugated AffiniPure goat anti‐mouse IgG (H+L) for 1 h at room temperature. Nuclei were counterstained with DAPI. Then the samples were mounted with clear liquid mounting medium. Fluorescence signals were observed using a confocal laser scanning microscope N‐STORM & A1 (Nikon, Tokyo, Japan) and analyzed with NIS analysis software.

Huang L., Sun X., Wang L., Pei G., Wang Y., Zhang Q., Liang Z., Wang D., Fu C., He C, & Wei Q. (2022). Enhanced effect of combining bone marrow mesenchymal stem cells (BMMSCs) and pulsed electromagnetic fields (PEMF) to promote recovery after spinal cord injury in mice. MedComm, 3(3), e160.

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Mitochondrial Morphology Analysis of MSCs

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MSCs were stained with 50 nM MitoTracker Green and Hoechst 33258 (Invitrogen, USA) at 37 °C for 30 min. After washing with PBS, representative images were observed via confocal microscopy (Nikon, N-STORM & A1, Tokyo, Japan). The aspect ratio (AR, major axis/minor axis) and form factor (FF, 4π × (area/perimeter²)) were calculated using ImageJ software (NIH) as previously reported 12 (link).

Zhao M., Liu S., Wang Y., Lv K., Lou P., Zhou P., Zhu J., Li L., Cheng J., Lu Y, & Liu J. (2024). The mitochondria‒paraspeckle axis regulates the survival of transplanted stem cells under oxidative stress conditions. Theranostics, 14(4), 1517-1533.

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