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Fluoview 3000 confocal microscope

Manufactured by Olympus
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The Fluoview 3000 confocal microscope is a high-performance imaging system designed for advanced fluorescence microscopy applications. It features a modular and flexible design that allows for customization to suit various research needs. The Fluoview 3000 is capable of capturing high-resolution, multi-dimensional images with exceptional image quality.

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Lab products found in correlation

Vectashield mounting medium, by Vector Laboratories (3 mentions) Lsm 780 confocal microscope, by Zeiss (3 mentions) Fluoview fv10 asw 4.0 viewer software, by Olympus (2 mentions) Fv10 asw 3, by Olympus (2 mentions) Prism software, by GraphPad (2 mentions) Fluoview fv10 asw 4.1 software package, by Olympus (2 mentions) Prolong gold antifade reagent with dapi, by Cell Signaling Technology (2 mentions) Ms 222, by Merck Group (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Alexa fluor 568, by Thermo Fisher Scientific (2 mentions) 20r lr002, by Biosynth (1 mentions) Filipin 3, by Merck Group (1 mentions) Rhodamine 123, by Merck Group (1 mentions) Q5 camera, by Olympus (1 mentions) Fitc pna, by Merck Group (1 mentions) Bx51 fluorescence microscope, by Olympus (1 mentions) Neutral buffered formalin, by Merck Group (1 mentions) Alexa fluor 594 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Click it edu imaging kit, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Fluoview 3000 Laser Scanning Confocal Microscope Fluoview 3000 inverted confocal microscope Fluoview 3000 microscope FluoView confocal microscope Fluoview 3000 Confocal microscope

24 protocols using fluoview 3000 confocal microscope

1

Immunostaining of LDLR in Cells

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Cells were transfected and cultured as described above then fixed with 3% paraformaldehyde (PFA) at room temperature for 20 min, washed with PBS, incubated with 30 mM glycine for 5 min and washed again with PBS. For LDLR staining cells were permeabilized with 0.05% Filipin III (Sigma #F4767) in 10% FCS for 30 min at room temperature. Primary antibody: rabbit monoclonal anti-LDLR (Fitzgerald #20R-LR002) was diluted 1:100 in 5% FCS overnight at 4 °C. Secondary antibody: goat polyclonal goat anti-rabbit IgG Alexa 568 (Invitrogen #A11011) was diluted 1:400 in 5% FCS. Fixed cells were imaged using a Zeiss LSM 780 confocal microscope using a 63×/1.4NA oil immersion objective. For plasma membrane LDLR staining, cells were not permeabilized, but directly stained for 1 hour at 4 °C with antibody recognising the extracellular part of LDLR, namely, mouse anti-LDLR-C7 antibody (Progen #61087) diluted 1:100 in PBS. As a secondary antibody, we used chicken anti-mouse IgG Alexa 488 (Invitrogen #A-21200) diluted 1:400. Cells were imaged on an Olympus FluoView 3000 confocal microscope using 60×1.3NA silicon oil immersion objective.
Zimoń M., Huang Y., Trasta A., Halavatyi A., Liu J.Z., Chen C.Y., Blattmann P., Klaus B., Whelan C.D., Sexton D., John S., Huber W., Tsai E.A., Pepperkok R, & Runz H. (2021). Pairwise effects between lipid GWAS genes modulate lipid plasma levels and cellular uptake. Nature Communications, 12, 6411.
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2

Sperm Acrosome and Mitochondrial Integrity

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Sperm acrosome integrity was assessed using a fluorescent-labeled peanut agglutinin (FITC-PNA) (Sigma-Aldrich) as described previously [15 (link)]. Briefly, smears of sperm suspension were fixed in methanol for 15 min, air-dried, and stained with FITC-PNA (60 μg/ml in PBS) in the dark for 30 min. Slides were then washed with Milli-Q water to remove excessive staining. Two hundred spermatozoa were evaluated under Olympus Fluoview 3000 confocal microscope at 600× magnification and classified as intact or damaged acrosome based on the staining since FITC-PNA binds exclusively to the outer acrosomal membrane.
Sperm mitochondrial activity was assessed using a fluorescent dye Rhodamine 123 that is rapidly taken up by functional mitochondria. In brief, 5 μl of Rhodamine 123 (Sigma, U.S.A.) solution (1 mg/ml in DMSO) was added to 1 ml of diluted sperm suspension and incubated for 10 min at 25°C in the dark. Following incubation, the supernatant was removed by centrifugation (300× g, 10 min) and the sperm pellets were resuspended in 1 ml PBS. A drop of 10 μl of the suspension was placed on microscopic slides, covered with coverslips, and examined at 400× magnification under Olympus BX51 fluorescence microscope with Olympus Q5 camera. A total of 200 spermatozoa were examined per animal and expressed as percentage of mitochondrial activity.
Navaneethabalakrishnan S., Wilcox B.K., Goodlett B.L., Murphy M.M, & Mitchell B.M. (2022). Hypertension induces gonadal macrophage imbalance, inflammation, lymphangiogenesis, and dysfunction. Clinical science (London, England : 1979), 136(11), 879-894.
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3

Intracellular Trafficking of Exendin-4 in Caco-2 Cells

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Caco-2 cells were seeded in an eight-well chamber slide and incubated with 100 nM Alexa 594–LCFA-Ex4 for 1 hour. LysoTracker Green DND-26, CellLight Early Endosomes–GFP, CellLight Late Endosomes–GFP, or ER-Tracker Green was added following manufacturer-suggested protocols. Cells were fixed with 4% paraformaldehyde and then stained with 4′,6-diamidino-2-phenylindole (DAPI). Slides were mounted, and confocal images were taken using a FluoView 3000 confocal microscope from Olympus.
Hu Z., Nizzero S., Goel S., Hinkle L.E., Wu X., Li C., Ferrari M, & Shen H. (2020). Molecular targeting of FATP4 transporter for oral delivery of therapeutic peptide. Science Advances, 6(14), eaba0145.
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4

Immunofluorescence Staining of Cultured Cells

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After indicated treatments, cells were pre-extracted with CSK buffer (10 mM PIPES, pH 6.8, 100 mM NaCl, 300 mM sucrose, 3 mM MgCl2, 1 mM EGTA, 0.5% [v/v] Triton X-100) on ice for 5 min and were fixed with 2% paraformaldehyde (PFA) at room temperature for 15 min. After washing with PBS, cells were blocked with 3% BSA in TBST for 30 min and incubated with the indicated primary antibodies at 4°C for 18 h. Cells were then washed 3× in PBS and stained with the appropriate secondary antibody at room temperature for 1 h. Cover slips were mounted onto 1.2-mm glass slides using Vectashield mounting medium containing DAPI (Vector Labs) and analyzed by FV10-ASW3.1 software on a Fluoview 3000 confocal microscope (Olympus).
Kim J.J., Lee S.Y., Gong F., Battenhouse A.M., Boutz D.R., Bashyal A., Refvik S.T., Chiang C.M., Xhemalce B., Paull T.T., Brodbelt J.S., Marcotte E.M, & Miller K.M. (2019). Systematic bromodomain protein screens identify homologous recombination and R-loop suppression pathways involved in genome integrity. Genes & Development, 33(23-24), 1751-1774.
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5

Immunofluorescence Staining of Tissue Samples

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The tissues and explants were fixed for 24 h in 10% neutral-buffered formalin (Sigma-Aldrich) at room temperature. For immunofluorescence, CD105 polyclonal (Thermo Fisher, PA5-94980), CD3 monoclonal (F7.2.38) (Thermo Fisher, MA5-12577), Sox2 polyclonal (Thermo Fisher, 48-1400), GFAP monoclonal (ASTRO6) (Thermo Fisher, MA5-12023), GFAP polyclonal (Abcam, ab7260), Iba1 polyclonal (Wako 091-19741) primary antibodies were then applied at dilutions of 1:400, 1:20, 1:100, 1:100, 1:250, 1:1000, respectively, overnight at 4°C, and visualized using Olympus Fluoview 3000 confocal microscope at a magnification of 60X. See supplementary materials for more information.
Morelli M., Lessi F., Barachini S., Liotti R., Montemurro N., Perrini P., Santonocito O.S., Gambacciani C., Snuderl M., Pieri F., Aquila F., Farnesi A., Naccarato A.G., Viacava P., Cardarelli F., Ferri G., Mulholland P., Ottaviani D., Paiar F., Liberti G., Pasqualetti F., Menicagli M., Aretini P., Signore G., Franceschi S, & Mazzanti C.M. (2022). Metabolic-imaging of human glioblastoma live tumors: A new precision-medicine approach to predict tumor treatment response early. Frontiers in Oncology, 12, 969812.
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6

Visualizing MsSPL26 and MsSPL13 Localization

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The coding region of MsSPL26 without termination codons was amplified and inserted into the NcoI/BglII sites of the pCAMBIA1302 vector, resulting in the MsSPL26-GFP fusion construct. The MsSPL13-GFP and MsSPL26-GFP constructs were transformed into A. tumefaciens strain GV3101 and transiently expressed in N. benthamiana leaves. The fluorescence signal was detected after 2 d of incubation using an Olympus FluoView 3000 confocal microscope equipped with Olympus FluoView FV10-ASW 4.0 Viewer Software. As for visualizing the nucleus, 30 μM DAPI was used to stain the nucleus of the epidermal cells in N. benthamiana leaves for 10 min at 37 °C. GFP was excited at 488 nm and the emitted signal was captured at 500 to 540 nm. DAPI was excited at 408 nm and the emitted signal was captured at 425 to 475 nm. Images were captured using 5% of the maximum light intensity value and gain of 600 to 650.
Feng C., Zhang X., Du B., Xiao Y., Wang Y., Sun Y., Zhou X., Wang C., Liu Y, & Li T.H. (2023). MicroRNA156ab regulates apple plant growth and drought tolerance by targeting transcription factor MsSPL13. Plant Physiology, 192(3), 1836-1857.
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7

Quantifying DNA Repair Foci in U2OS Cells

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U2OS cells were transfected with indicated siRNAs and incubated for 48 h. Cells were then treated with IR (5 Gy) and immunofluorescence (IF) analysis was performed after 2 h further incubation in tissue culture incubator (37°C, 5% CO2). Cells were pre-extracted with 1 ml of CSK buffer containing 10 mM PIPES, pH 6.8, 100 mM NaCl, 300 mM sucrose, 3 mM MgCl2, 1 mM EGTA and 0.5% (v/v) Triton X-100 on ice for 5 min followed by fixation with 2% paraformaldehyde (PFA) for 15 min at RT. Subsequently, cells were washed 3 times with PBS (1 ml) and blocked with 1 ml of 3% BSA in TBS-T (0.05% Tween-20) for 30 min. RAD51 antibody (GeneTex, #GTX100469) was incubated for 18 h at 4°C and then washed with 1 ml of PBS (10 min, 3 times) at RT. After washing, cells were incubated with Alexa Fluor 594 goat anti-rabbit IgG (Invitrogen, #A-11012) for 1 h at RT and mounted onto 1.2 mm glass slide using DAPI containing Vectashield mounting medium (Vector laboratories, #H-1200). Samples were analyzed using a Fluoview 3000 confocal microscope (Olympus) and the number of RAD51 foci per cell was counted.
Kim J.J., Lee S.Y., Choi J.H., Woo H.G., Xhemalce B, & Miller K.M. (2020). PCAF-mediated histone acetylation promotes replication fork degradation by MRE11 and EXO1 in BRCA-deficient cells. Molecular cell, 80(2), 327-344.e8.
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8

Laser-Induced DNA Damage Imaging

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Laser micro-irradiation and live-cell imaging methods were performed as described (Kim et al., 2019a (link)). Briefly, cells were seeded on glass-bottomed dishes (Ted Pella) and pre-sensitized with 10 μM BrdU for 24 h before laser-induced damage. Cells were visualized with 60X oil objective lens and laser-induced DNA damages were generated by a 405-nm laser beam (60% laser intensity) using a Fluoview 3000 confocal microscope (Olympus). During laser damage and live-cell imaging, cells were maintained in an environmental chamber (37°C, 5% CO2). All experiments were analyzed using FV-10-ASW3.1 software (Olympus). Fluorescence intensity at damage sites was calculated by comparing the intensity of damaged versus non-damaged sites within the same cell as a function of time. For each group, at least 10 individual cells were analyzed and quantification results were graphed using Prism software (GraphPad).
Kim J.J., Lee S.Y., Choi J.H., Woo H.G., Xhemalce B, & Miller K.M. (2020). PCAF-mediated histone acetylation promotes replication fork degradation by MRE11 and EXO1 in BRCA-deficient cells. Molecular cell, 80(2), 327-344.e8.
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9

Immunofluorescent Staining of FFPE Brain Tissues

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Formalin-fixed, paraffin-embedded (FFPE) tissue blocks, obtained from our GB samples, were cut into 2–4 μm thick sections. Antigen unmasking was achieved with Epitote Retrieval Solution (pH = 8) (Leica Microsystems, Wetzlar, Germany) in a microwave. GFAP monoclonal (ASTRO6) (MA5-12023, Thermofisher Scientific, Waltham, MA, USA) and IBA1 polyclonal (091-19741, Wako Chemicals, Neuss, Germany) primary antibodies were then applied at dilutions of 1:100 and 1:1000, respectively, overnight at 4 °C. The goat anti-rabbit Alexa Fluor 488 (Thermofisher Scientific, Waltham, MA, USA ) and goat anti-mouse Alexa Fluor 568 (Thermofisher Scientific, Waltham, MA, USA ) were diluted 1:500 and incubated for 1 h. Cells were counterstained with DAPI (Sigma Aldrich, St. Louis, MO, USA) and visualized using an Olympus Fluoview 3000 confocal microscope at a magnification of 60×.
Lessi F., Franceschi S., Morelli M., Menicagli M., Pasqualetti F., Santonocito O., Gambacciani C., Pieri F., Aquila F., Aretini P, & Mazzanti C.M. (2022). Single-Cell Molecular Characterization to Partition the Human Glioblastoma Tumor Microenvironment Genetic Background. Cells, 11(7), 1127.
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10

Chikungunya virus infection of BeWo and HUVEC cells

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Monolayers of BeWo and HUVEC cells were infected with 100 infectious units per well. After 48 h, samples were fixed with 4% paraformaldehyde and blocked with 5% lamb serum. Cells were stained with anti-CHIKV monoclonal antibody 3E7b and anti-MAP2 antibody (Novus Biologicals, Littleton, CO, USA). Slides were mounted with ProLong Gold Antifade Reagent with DAPI (Cell Signaling Technology, Danvers, MA, USA catalog #8961S) and images were obtained using an Olympus Fluoview 3000 confocal microscope. Images were processed using the Olympus Fluoview FV10-ASW 4.1 software package.
Schultz E.M., Jones T.J, & Barr K.L. (2020). Antibodies for Venezuelan Equine Encephalitis Virus Protect Embryoid Bodies from Chikungunya Virus. Viruses, 12(3), 262.
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