Fv1200 laser scanning confocal microscope

Manufactured by Olympus

Sourced in Japan, United States

The FV1200 is a laser scanning confocal microscope designed for high-resolution imaging of fluorescently-labeled samples. It features a range of laser excitation wavelengths, adjustable pinhole sizes, and sensitive photomultiplier tube (PMT) detectors to capture detailed optical sections of specimens.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (14 mentions) 35 mm glass dish, by Iwaki (6 mentions) Bx53 light microscope, by Olympus (4 mentions) Bovine serum albumin (bsa), by Merck Group (3 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (3 mentions) Fluoview software, by Olympus (3 mentions) Cm1850 cryostat, by Leica (3 mentions) μ slide 8 well, by Ibidi (3 mentions) Hoechst 33342 dye, by Thermo Fisher Scientific (3 mentions) Cellsens dimension 1, by Olympus (3 mentions) Texas red dextran, by Thermo Fisher Scientific (3 mentions) Hoechst 33342, by Thermo Fisher Scientific (3 mentions) Fluo 4 am, by Thermo Fisher Scientific (2 mentions) Fv10 asw software, by Olympus (2 mentions) Acetone, by Thermo Fisher Scientific (2 mentions) R9 kim 1 antibody, by Merck Group (2 mentions) Cyp2e1 antibody, by Merck Group (2 mentions) Rhs 1 microwave, by Milestone (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Cd8β af647 yts156, by BioLegend (2 mentions)

Close spelling variants of this product

FV1200 (803 citations) Confocal microscope (790 citations) Confocal laser scanning microscope (768 citations) FV1200 confocal microscope (264 citations) Laser confocal microscope (195 citations) FV1200 microscope (52 citations) Laser scanning microscope (29 citations) FLUOVIEW FV1200 confocal laser scanning microscope (29 citations) FV1200 Laser Scanning Microscope (28 citations) FV1200 biological confocal laser-scanning microscope (21 citations)

156 protocols using fv1200 laser scanning confocal microscope

Visualizing Cellular Dynamics with EV Treatment

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CHO-K1 cells (2 × 10⁵ cells, 2 mL) were plated onto a 35-mm glass dish (Iwaki, Tokyo, Japan) and incubated in Ham’s F-12 containing 10% FBS for 24 h at 37 °C under 5% CO₂. After complete adhesion, the cells were washed with cell culture medium containing 10% FBS and treated with each EV sample (100 μl/well). The cells were stained with Hoechst 33342 dye (Invitrogen; 5 μg/ml) for 15 min at 37 °C before cell washing. The cells were then washed with fresh cell culture medium and analysed using a FV1200 confocal laser scanning microscope (Olympus, Tokyo, Japan) equipped with a 40x objective without cell fixation.
To visualize the lamellipodia formation of cellular actin, the cells were fixed with 4% paraformaldehyde at room temperature for 30 min and washed with PBS after the cells were treated with each EV sample (100 μl/well). The cells were then treated with 0.1% Triton X-100 (100 μl/well in PBS) at room temperature for 5 min and again washed with PBS. Cellular F-actin was stained with rhodamine-phalloidin (2.5 μl (300 units) in PBS (97.5 μl)/well) (Molecular Probes) for 20 min at 4 °C, and the cells were washed with PBS before analysis using a FV1200 confocal laser scanning microscope (Olympus) equipped with a 40× objective.

Nakase I., Noguchi K., Aoki A., Takatani-Nakase T., Fujii I, & Futaki S. (2017). Arginine-rich cell-penetrating peptide-modified extracellular vesicles for active macropinocytosis induction and efficient intracellular delivery. Scientific Reports, 7, 1991.

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Immunostaining of Extracellular Vesicles and Tissue Samples

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Cells were seeded onto glass coverslips at the density of 5 × 10⁴ cells/ml in a two‐well confocal chamber, incubated overnight, and treated with sEVs or IL2‐sEVs for an additional 48 h. For detection, an antibody was added to the culture medium and incubated for 1 h before fixing with 4% PFA for 10 min at room temperature. The fixed cells were washed with PBS and the coverslips were stained and mounted using a DAPI‐containing mounting solution. Immunostaining of 5 μm thick specimens of O.C.T. compound‐embedded fresh frozen cancer tissues was performed using PD‐L1, Rab27a, and CD63 antibodies. After fixation with 4% PFA, nonspecific binding was blocked using 5% normal goat serum in PBS for 1 h. The slides were incubated with primary antibodies diluted in 5% normal goat serum (100 μl/slide) overnight at 4°C. After washing, secondary antibodies were added and incubated for 1 h. The slides were washed again, and the coverslip was stained and mounted using a DAPI‐containing mounting solution. Fluorescence images were obtained using an Olympus/FV1200 laser scanning confocal microscope.

Jung D., Shin S., Kang S., Jung I., Ryu S., Noh S., Choi S., Jeong J., Lee B.Y., Kim K., Kim C.S., Yoon J.H., Lee C., Bucher F., Kim Y., Im S., Song B., Yea K, & Baek M. (2022). Reprogramming of T cell‐derived small extracellular vesicles using IL2 surface engineering induces potent anti‐cancer effects through miRNA delivery. Journal of Extracellular Vesicles, 11(12), 12287.

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Immunofluorescent Localization of β-Catenin and NFATc1

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The cells were fixed in 4% formaldehyde and perm with 10% TritonX-100 for 15 min, washed three times with 0.01 M PBS, and incubated with 5% bovine serum albumin (Sigma-Aldrich) for 1 h for blocking. The sections were then incubated overnight at 4°C with anti-β-catenin (15B8, eBioscience) or anti-NFATc1 (7A6, Santa Cruz Biotechnology) primary antibodies. Next, the cells were washed three times with 0.01 M PBS, incubated with secondary antibody Alexa Fluor 488-conjugated Goat anti-Mouse IgG (#A11001, Invitrogen) for 4 h and washed three times with 0.01 M PBS. For nuclear counterstaining, the cells were incubated with 4′,6-diamidino-2-phenylindone (Cell Signaling Technology, #4083) for 10 min, washed in 1× PBS, and the slides were set with FluorSave™ reagent (EMD Millipore). Expression and localization of beta-catenin were observed with an Olympus FV1200 laser scanning confocal microscope (Olympus Corporation) with fixed exposure time for all samples. Colocalization was quantified as a ratio of overlapping pixels to the total number of pixels by Threshold Mander’s coefficient and expressed as nuclear percentage (% nuclear). The preparations were visualized using confocal microscopy (Olympus FV1200), and the analyses of images were carried out using FIJI/ImageJ.

Volpini X., Ambrosio L.F., Fozzatti L., Insfran C., Stempin C.C., Cervi L, & Motran C.C. (2018). Trypanosoma cruzi Exploits Wnt Signaling Pathway to Promote Its Intracellular Replication in Macrophages. Frontiers in Immunology, 9, 859.

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Quantifying Cofilin Nuclear Translocation

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Following stimulation with cyclic stretch, cells in BioFlex culture plates were fixed with 4% formaldehyde for 30 min at room temperature. The elastic bottom of each plate was cut into 8–10 pieces, which were then blocked in 2% goat serum (Beyotime Institute of Biotechnology, Haimen, China) in phosphate-buffered saline at 37°C for 1 h. Following blocking, the cells were incubated with monoclonal rabbit anti-active cofilin primary antibody (1:100; cat. no. 5175; Cell Signaling Technology, Inc.) for 1 h. Cells were then incubated with fluorescein isothiocyanate-conjugated goat anti-rabbit IgG secondary antibody (1:500; cat. no. 65–6111; Thermo Fisher Scientific, Inc.) for 1 h and counterstained with 4′,6-diamidino-2-phe-nylindole for 5 min at room temperature. Cells were then visualized with an FV1200 laser scanning confocal microscope (Olympus Corporation, Tokyo, Japan). Six fields of view were randomly selected and the number of cells exhibiting nuclear translocation of cofilin were counted and normalized to the total number of cells in each field.

GAO J., FU S., ZENG Z., LI F., NIU Q., JING D, & FENG X. (2016). Cyclic stretch promotes osteogenesis-related gene expression in osteoblast-like cells through a cofilin-associated mechanism. Molecular Medicine Reports, 14(1), 218-224.

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Quantifying Smad4 Nuclear Localization

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FRAP experiments were done on an Olympus FV1200 laser scanning confocal microscope using a 100x NA 1.49 oil objective. To deal with cell movement recovery, curves were obtained by reading out a polygon defined by the interpolation between the initial bleach window and a manually defined polygonal nuclear mask in the final frame. Supplemental Information describes the mathematical model for nuclear localization of Smad4 which is shown schematically in Figure 2g,h, and the inference of model parameters shown in Figure 2i,j from the data.

Heemskerk I., Burt K., Miller M., Chhabra S., Guerra M.C., Liu L, & Warmflash A. (2019). Rapid changes in morphogen concentration control self-organized patterning in human embryonic stem cells. eLife, 8, e40526.

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Cellular Calcium Imaging with Fluo-4-AM

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To measure cellular Ca²⁺ levels, Fluo-4-AM (Molecular Probes, Eugene, OR, USA) was used for cell staining. Cells (2.0 × 10⁵ cells/mL) were seeded on chamber slides and sequentially treated with esculetin and/or H₂O₂. The stained cells were imaged using the FV1200 laser scanning confocal microscope (Olympus, Tokyo, Japan).

Zhen A.X., Piao M.J., Kang K.A., Fernando P.D., Kang H.K., Koh Y.S, & Hyun J.W. (2019). Esculetin Prevents the Induction of Matrix Metalloproteinase-1 by Hydrogen Peroxide in Skin Keratinocytes. Journal of Cancer Prevention, 24(2), 123-128.

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Immunolabeling of Corneal Basement Membrane

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Immunolabeling was performed on five of the corneal rims fixed in 4% paraformaldehyde. The tissue was bisected and relief cuts were made in the tissue to allow it to flatten out. The tissue was permeabilized with PBS-Tx overnight at 4C, then brought to room temperature and incubated in sodium borohydride, on a rocker, for ten minutes and rinsed in phosphate-buffered saline (PBS) (2 min x4). While still on the rocker, tissue was incubated 1 hr at RT in blocking buffer (containing 0.3% Triton X-100 and 10% heat-inactivated donkey serum ), then washed with PBS and incubated for four hours with primary mouse monoclonal anti-human type VII collagen antibody 5D2 (from SundarRaj, University of Pittsburgh, Pittsburgh, PA) and washed with blocking buffer (10 min x5) prior to 1 hr incubation with Alex-Fluor 488-conjugated donkey anti-mouse IgG secondary antibody, and washed with PBS (10 min x4). Finally, tissue was incubated in Hoechst and then washed in PBS (15min x4). The corneal rim halves were placed in a glass dish with a spacer and sealed with a glass coverslip. Stitched stack volumes were acquired and reconstructed with an Olympus FV1200 laser scanning confocal microscope with mosaic scanning and a 20x UPlan S-APO lens (numerical aperture 0.85) Fluoview software and median stack projections were built in FIJI to show the pattern of the basement membrane with FIJI software.

Sigal I.A., Steele J., Drexler S, & Lathrop K.L. (2016). Identifying the Palisades of Vogt in Human Ex-vivo Tissue. The ocular surface, 14(4), 435-439.

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Olympus Confocal Microscopy Protocol

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All imaging was done using an Olympus FV1200 Laser Scanning Confocal Microscope (60 × oil objective with 1.4 NA or 40 × silicone objective with 1.25 NA) and Flow View software.

Boucher J.M., Clark R.P., Chong D.C., Citrin K.M., Wylie L.A, & Bautch V.L. (2017). Dynamic alterations in decoy VEGF receptor-1 stability regulate angiogenesis. Nature Communications, 8, 15699.

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Intracellular ROS Detection in HaCaT Cells

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For the detection of intracellular ROS in HaCaT cells, the cells were seeded in 6-well culture plates at a density of 1.0 × 10⁵ cells/well, cultured for 16 h, and treated with 20 μg/mL FFO and 50 μg/mL PM_2.5. After 30 min, 50 μM 2’,7’-dichlorodihydrofluorescein diacetate (DCF-DA, Molecular Proves, Eugene, OR, USA) solution was added to the cells. After 15 min incubation at 37 °C, DCF fluorescence was measured using a BD LSRFortessa flow cytometer (PerkinElmer, Waltham, MA, USA). For imaging, cells were seeded in a 4-well glass chamber slide at a density of 1.0 × 10⁵ cells/well, cultured for 16 h, and treated with 20 μg/mL FFO and 50 μg/mL PM_2.5. After 30 min, 50 μM DCF-DA (Molecular Proves, Eugen, OR, USA) solution was added to the cells. After 15 min incubation at 37 °C, the cells were washed 3 times with PBS. DCF fluorescence images were obtained using a FV1200 laser scanning confocal microscope (Olympus, Tokyo, Japan).

Hyun Y.J., Piao M.J., Kang K.A., Zhen A.X., Madushan Fernando P.D., Kang H.K., Ahn Y.S, & Hyun J.W. (2019). Effect of Fermented Fish Oil on Fine Particulate Matter-Induced Skin Aging. Marine Drugs, 17(1), 61.

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Quantifying Proliferating Cells in Mice

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1 mg of EdU (dissolved in water) was injected intraperitoneally into mice 1 h before euthanasia. Mice were perfused with 5% NBF and the meninges were removed as described above. EdU was detected using a Click-iT Plus EdU Alexa Fluor 647 kit (ThermoFisher) and was imaged with an Olympus FV1200 laser scanning confocal microscope. Quantification of EdU⁺ cells was performed by histo-cytometry as described below.

Russo M.V., Latour L.L, & McGavern D.B. (2018). Distinct myeloid cell subsets promote meningeal remodeling and vascular repair after mild traumatic brain injury. Nature immunology, 19(5), 442-452.

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