Fv1000 confocal microscope system

Manufactured by Olympus

Sourced in Japan

The FV1000 confocal microscope system is a high-performance imaging platform designed for advanced fluorescence microscopy applications. It features a modular architecture, allowing for customization to meet specific research requirements. The system enables high-resolution, optical sectioning of samples, providing detailed visualization of cellular and subcellular structures.

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FV1000 (3240 citations) FV1000 confocal microscope (1548 citations) Confocal microscope (790 citations) FV1000 microscope (233 citations) FV1000 confocal system (65 citations) FV1000 system (38 citations) Confocal FV1000 (10 citations) Olympus System Microscope (4 citations)

20 protocols using fv1000 confocal microscope system

Axl Protein Detection in Tumor Cells

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Whole cell protein lysate was obtained by enhanced RIPA buffer (Beyotime). Samples were sonicated and then centrifuged at 15,000 g for 10 min at 4°C. Protein concentrations were determined by Bradford assay (Beyotime). Equal amounts of protein were fractionated by SDS-PAGE, transferred to a PVDF membrane (Millipore), and analyzed by incubation with primary rabbit anti-mouse/human Axl antibody. Proteins were detected via incubation with HRP-conjugated secondary antibodies and enhanced chemiluminescence (ECL) detection system (Cell Signaling Technology).
For immunofluorescence staining, tumor cells were seeded on collagen-coated glass slides and grown to 50–60% confluence. Cells were then fixed in fresh 3% formaldehyde in PBS followed by permeabilization and blocking in Tris-buffered saline + 0.05% Tween 20 (TBST) containing 5% bovine serum albumin and 0.3% Triton X-100 at room temperature. After that, cells were stained with primary rabbit anti-mouse/human Axl antibody followed by secondary FITC-labeled goat anti-rabbit antibody. After final washes in TBST, cells were mounted in ProLong Gold anti-fade reagent (Life Technologies) and imaged on the Olympus FV1000 confocal microscope system. Acquired images were processed by ImageJ software.

Guo Z., Li Y., Zhang D, & Ma J. (2017). Axl inhibition induces the antitumor immune response which can be further potentiated by PD-1 blockade in the mouse cancer models. Oncotarget, 8(52), 89761-89774.

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Confocal Microscopy of Spinal Cord

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Samples were coverslipped with Vectashield (Vector Laboratories), and all images were acquired at room temperature. Automated confocal scanning of spinal cord tissue was performed with the FV10-ASW microscopy software on an upright Olympus FV1000 confocal microscope system. Images were acquired using standard filter sets, and acquisition settings were kept constant between control and treatment groups for each experiment.

Bradley P.M., Denecke C.K., Aljovic A., Schmalz A., Kerschensteiner M, & Bareyre F.M. (2019). Corticospinal circuit remodeling after central nervous system injury is dependent on neuronal activity. The Journal of Experimental Medicine, 216(11), 2503-2514.

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Salmonella Invasion Assay in J774 Cells

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Control and ELMO1 shRNA (J774) cells were infected with RFP-labeled Salmonella (SL1344-RFP) for 5 minutes at a moi of 10. Samples were washed in 1X PBS, pH 7.4 and fixed in 2% formaldehyde, washed with PBS, and permeabilized with 0.1% triton in PBS for 5 minutes. Cells were blocked with 5% goat serum -1.5% bovine serum albumin in PBS (blocking solution) and subsequently incubated with Alexa 488 Phalloidin (Life Technologies) to stain F- actin (green).
The cells were washed and incubated for 5 minutes in Hoechst 33342 (Life Technologies) diluted 1:2000 in 1X PBS to stain the nuclei. The cells were washed with 1X PBS and subsequently mounted on glass slides with prolong gold. Confocal images were obtained with a 100× objective using an Olympus FV1000 Confocal microscope system (Olympus America, Center Valley, PA).

Das S., Sarkar A., Choudhury S.S., Owen K.A., Derr-Castillo V.L., Fox S., Eckmann L., Elliott M.R., Casanova J.E, & Ernst P.B. (2015). Engulfment and Cell Motility Protein 1 (ELMO1) Has an Essential Role in the Internalization of Salmonella Typhimurium Into Enteric Macrophages That Impact Disease Outcome. Cellular and Molecular Gastroenterology and Hepatology, 1(3), 311-324.

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Apoptosis, Necrosis, and Cell Viability Assay

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Cells were stained with an apoptotic/necrotic/health cell detection kit (PromoCell GmbH, Heidelberg, Germany) containing FITC-Annexin V, Ethidium Homodimer III (EthD-III), and Hoechst 33342. In brief, cultured cells on the glass slide were washed using a binding buffer and stained with the staining solution for 15 min. Subsequently, the cells were washed twice with the binding buffer and observed under confocal microscopy (40× objective) using the Olympus FV1000 confocal microscope system (Olympus, Tokyo, Japan). The observed cells were in an unfixed state.

Miura K, & Fukushi Y. (2021). Scanning acoustic microscopy imaging of cellular structural and mechanical alterations from external stimuli. Heliyon, 7(8), e07847.

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Quantifying Acetylated p65 in BMMs

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BMMs were plated onto a glass coverslip in RPMI 1640 supplemented with 10% FBS and incubated overnight at 37°C. The following day, the cells were treated with TNF-α (10 ng/ml) for 1 h. At the end of the treatments, the cells were washed with PBS, fixed in PBS containing 4% paraformaldehyde, and permeabilized with 0.1% Triton X-100 in PBS for 10 min at room temperature. Then, the cells were blocked with 2% bovine serum albumin (BSA) in PBS for 1 h and incubated with primary monoclonal antibodies against acetylated p65 overnight at 4°C. After washing in PBS, the cells were further incubated with TRITC-conjugated secondary anti-mouse IgG for 1 h. The nuclei were stained with DAPI (0.5 µg/mL; Invitrogen-Molecular Probes). The cover slips were mounted and stored in the dark at 4°C. Fluorescent images were obtained from analyses using the FV-1000 confocal microscope system (Olympus, Tokyo, Japan).

Hah Y.S., Cheon Y.H., Lim H.S., Cho H.Y., Park B.H., Ka S.O., Lee Y.R., Jeong D.W., Kim H.O., Han M.K, & Lee S.I. (2014). Myeloid Deletion of SIRT1 Aggravates Serum Transfer Arthritis in Mice via Nuclear Factor-κB Activation. PLoS ONE, 9(2), e87733.

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Fluorescence Microscopy of Meiotic Chromosomes

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All images of meiotic chromosomes after silver staining and immunodetection of 5-MeC and sequential FISH experiments were acquired using a Zeiss Axio Imager.Z.2 wide-field fluorescence microscope equipped with an AxioCam HRm monochromatic camera. Meiocytes embedded in polyacrylamide and embryo sections were optically sectioned using an Olympus FV1000 confocal microscope system equipped with a ×60/1.35 PlanApo objective. Image stacks were acquired by traversing from the top to the bottom of a nucleus in 0.2 µm steps. Image processing, including the rendering of the Z-stacks from a series of optical sections of the meiocytes, was performed using ImageJ (https://imagej.nih.gov/ij/; last accessed 10/12/2018).

Borowska-Zuchowska N., Robaszkiewicz E., Wolny E., Betekhtin A, & Hasterok R. (2018). Ribosomal DNA loci derived from Brachypodium stacei are switched off for major parts of the life cycle of Brachypodium hybridum. Journal of Experimental Botany, 70(3), 805-815.

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HMGB1 Effect on VE-Cadherin Localization

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Cells were grown on petri dishes and treated with HMGB1. Cells were then fixed in cold paraformaldehyde followed by extraction in 0.2% Triton X-100. Distribution of VE-cadherin was detected using rabbit anti-human VE-cadherin antibody and FITC-labeled goat anti-rabbit antibody as the secondary antibody. After subsequent washes, cells were incubated with DAPI at room temperature for 5 min. Cells were mounted in anti-photobleaching medium. Images were analyzed with a FV1000 confocal microscope system (Olympus, Tokyo, Japan). All pictures are representative of three independent experiments.

Zou M., Dong H., Meng X., Cai C., Li C., Cai S, & Xue Y. (2015). Store-Operated Ca2+ Entry Plays a Role in HMGB1-Induced Vascular Endothelial Cell Hyperpermeability. PLoS ONE, 10(4), e0123432.

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Fluorescence Microscopy for Metaphase Chromosome and Meiocyte Imaging

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Images of the mitotic metaphase chromosomes after FISH were acquired using a Zeiss Axio Imager.Z.2 wide-field fluorescence microscope equipped with an AxioCam HRm monochromatic camera. The meiocytes that had been embedded in the polyacrylamide gel were optically sectioned using an Olympus FV1000 confocal microscope system equipped with a 60×/1.35 PlanApo objective. All of the image stacks were acquired by scanning from the top to the bottom of a meiocyte in 0.25 μm steps and then processed using MBF ImageJ (Wayne Rasband, National Institutes of Health, Bethesda, MD, United States).

Borowska-Zuchowska N., Robaszkiewicz E., Mykhailyk S., Wartini J., Pinski A., Kovarik A, & Hasterok R. (2021). To Be or Not to Be Expressed: The First Evidence of a Nucleolar Dominance Tissue-Specificity in Brachypodium hybridum. Frontiers in Plant Science, 12, 768347.

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Immunostaining of Cultured Neurons

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Cultured neurons were briefly washed in cold PBS and fixed immediately with 4% paraformaldehyde in phosphate buffer saline (PBS) for 10 minutes. The formaldehyde fixed cells were incubated with permeablization buffer (0.5% Triton X-100, 100 mM glycine, 1% BSA, 0.7 mM EDTA) for 10 min on ice. The samples were then incubated with blocking buffer (10% bovine serum, 0.01% sodium azide in 1X PBS) at 37^oC for 1 hr or 4^oC overnight. Diluted primary antibodies (TUJ1, Sigma-Aldrich, 1:500; GLP-1R, Santa Cruz, 1:500) were incubated with samples for 1 hr at 37^oC. The samples were then rinsed with wash buffer (0.05% Tween-20 in 1xPBS) three times for 5 minutes at room temperature. A fluorophore conjugated secondary antibody (Alexa series, Invitrogen; 1:1,000 dilution), was added and the samples were incubated for 1 hr at 37^oC. Samples were rinsed briefly with wash buffer (3 x 5 minute) and mounted with Prolong Gold containing DAPI (nuclear marker, Invitrogen). Images of immunostained neurons were captured using a Olympus FV1000 confocal microscope system.

Yang J.L., Chen W.Y., Chen Y.P., Kuo C.Y, & Chen S.D. (2016). Activation of GLP-1 Receptor Enhances Neuronal Base Excision Repair via PI3K-AKT-Induced Expression of Apurinic/Apyrimidinic Endonuclease 1. Theranostics, 6(12), 2015-2027.

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Quantifying Cortical Neuronal Distribution

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Immunohistochemistry staining images were acquired using Olympus VS120 Automated Slide Scanner. Images for quantitative analyses were acquired using Olympus FV1000 confocal microscope system. Cell counting was performed on Z-stack confocal images. Briefly, the confocal images were imported into Photoshop CC (Adobe Systems Incorporated). We divided the human dorsal and dorsal-lateral neocortex into different number of bins with equal size (250 μm length and 125 μm width in 60 μm thick sections) spanning the cortical VZ, ISVZ (inner subventricular zone), IFL (inner fiber layer) [41 (link)], OSVZ [42 (link)], IZ (intermediate zone), SP (subplate), CP (cortical plate) and MZ (marginal zone). Cortical layers were delineated by DAPI staining. Labeled cells within each bin were manually counted by using a cell-counter and switching between the red, green, and blue channels in Photoshop. Three representative images of the dorsal or dorsal-lateral neocortex from randomly sampled sections were counted at each age (GW18 and GW23).

Yang L., Li Z., Liu G., Li X, & Yang Z. (2021). Developmental Origins of Human Cortical Oligodendrocytes and Astrocytes. Neuroscience Bulletin, 38(1), 47-68.

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