Fluoview 300

Manufactured by Olympus

Sourced in Japan, United States, Germany, United Kingdom

The Fluoview 300 is a laser scanning confocal microscope system designed for high-resolution fluorescence imaging. It features a compact design and supports a range of objective lenses, enabling detailed visualization of biological samples.

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Lab products found in correlation

R3896, by Hamamatsu Photonics (2 mentions) Uplansapo, by Olympus (2 mentions) Ix70 inverted laser scanning confocal microscope, by Olympus (2 mentions) Victor3, by PerkinElmer (2 mentions) Bx51 microscope, by Olympus (2 mentions) H2dcfda, by Thermo Fisher Scientific (2 mentions) Thunder imager, by Leica (2 mentions) Stellaris 8, by Leica (2 mentions) Fluoview software, by Olympus (2 mentions) Bes h2o2, by Fujifilm (2 mentions) Bes so, by Fujifilm (2 mentions) Confocal microscope, by Olympus (2 mentions) Phrodo staphylococcus aureus bioparticles conjugate, by Thermo Fisher Scientific (2 mentions) Fv300 laser scanning confocal microscope, by Olympus (2 mentions) Di02 r405, by IDEX Corporation (1 mentions) Lumplfln 60xw, by Olympus (1 mentions) Alexa fluor 488 or 594 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions) Fluorescein isothiocyanate (fitc), by Merck Group (1 mentions) Image pro plus 6, by Media Cybernetics (1 mentions)

Close spelling variants of this product

FluoView™ 300 confocal microscope (36 citations) Fluoview 300 fluorescence microscope (5 citations) Fluoview 300 confocal laser scanning microscope (5 citations) FluoView 300 laser scanning confocal head (3 citations) Fluoview FV 300 confocal system (3 citations) FluoView 300, Version 4.3 (2 citations) Fluoview 300 confocal laser scanning microscopy (2 citations) Fluoview 300 confocal unit (2 citations) Fluoview 300 microscope (2 citations) Fluoview 300 laser scanning microscope (2 citations)

79 protocols using fluoview 300

Simultaneous SHG and TPF Imaging

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SHG and TPF were excited using the 810 nm output of a Ti:Sapphire laser (Mira 900 Coherent) with 100fs pulses and a 76 MHz repetition rate.
For the tensile loading experiments, we imaged with a modified, non-inverted confocal laser scanning microscope (FluoView 300 and BX51 Olympus UK) with a 1NA 60x long working distance dipping lens (LUMPLFLN 60XW Olympus UK). For the compressive loading experiments, we imaged with a modified inverted confocal laser scanning microscope (FluoView 300 and IX71 Olympus UK) with a 1.2NA 60x objective (UPlanSApo Olympus UK).
For both experiments, SHG and TPF were collected simultaneously in the epi-direction. The signal was separated from the laser fundamental by a long pass dichroic filter (670dcxr Chroma technologies) and a colour glass filter (CG-BG-39 CVI laser) and the SHG and TPF were directed onto two separate photomultiplier tubes (PMTs) (R3896 Hamamatsu Japan) by a long pass dichroic filter (Semrock Di02-R405). Additional filters were placed in front of the PMTS (Semrock FF01-405/10 and Semrock FF01-520/70 for SHG and TPF respectively).
To avoid photodamage the average laser power at the sample was maintained below 20 mW.

Mansfield J.C., Bell J.S, & Winlove C.P. (2015). The micromechanics of the superficial zone of articular cartilage. Osteoarthritis and cartilage, 23(10).

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Two-Photon Imaging of Neurons and MCs

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Two-photon imaging of neurons and MCs in the IPL was performed using either a custom-modified two-photon microscope (Fluoview 300, Olympus America, Melville, NY) or a custom-built two-photon microscope. Time series images were acquired using Olympus 60×, 1 NA, LUMPlanFLN objectives, and two-photon excitation of GCAMP3 was evoked with an ultrafast pulsed laser (Chameleon Ultra, Coherent, Santa Clara, CA) tuned to 920 nm on both microscopes. For imaging Cal520 (Figure 6) the laser was tuned to 820 nm. The microscope was controlled by Fluoview Viewer software or ScanImage software (version 3.8, www.scanimage.org). Images (256 × 256 pixels) were acquired at 0.74 or 1.7 Hz at 2 or 4 ms/line. Scan parameters were [pixels/line × lines/frame (frame rate in Hz)]: [256 × 256 (0.74 − 1.7)], at 2–4 ms/line. Line scans were obtained at 300 Hz and down-sampled to 30 Hz for presentation in Figure 1—figure supplement 1.

Rosa J.M., Bos R., Sack G.S., Fortuny C., Agarwal A., Bergles D.E., Flannery J.G, & Feller M.B. (2015). Neuron-glia signaling in developing retina mediated by neurotransmitter spillover. eLife, 4, e09590.

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Trigeminal Ganglia Ca2+ Imaging

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Trigeminal ganglia excised from mice expressing the Ca²⁺ indicator GCaMP3 in neurons or glia were evaluated. Ca²⁺ transients were recorded from trigeminal ganglia using a confocal laser scanning microscope, Olympus Fluoview 300 equipped with 40× water-immersion lens (0.80 NA) and camera, filter set (emission/ excitation), software. Images were acquired at a rate of 40 frames/min. Excitation light was provided by an argon (488 nm) laser (Melles Girot, Carlsbad, CA). Ca²⁺ transients were quantified and expressed as the percentage of active cells within the field of view and as the frequency of Ca²⁺ spikes per min using ImageJ on selected regions placed on satellite glia and neurons of trigeminal ganglia derived from saline- and CFA-injected mice. Similar analyses were performed on ganglia exposed to ATP (10 and 30 μM).

Hanstein R., Hanani M., Scemes E, & Spray D.C. (2016). Glial pannexin1 contributes to tactile hypersensitivity in a mouse model of orofacial pain. Scientific Reports, 6, 38266.

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Immunofluorescence Staining of Osteogenic Markers

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The cells were washed with PBS, fixed for 5 min using 4% paraformaldehyde, and permeated for 20 min using 0.1% Triton™ X-100 at room temperature. The cells were then washed three times and blocked for 1 h using 4% BSA in PBS at room temperature. The cells were treated with primary antibodies (1:100; rabbit anti-OCN, rabbit anti-OSX, rabbit anti-Runx2) and incubated overnight at 4 °C. Subsequently, the cells were treated with Alexa Fluor® 488 or 594 goat anti-rabbit IgG (1:500; Invitrogen Life Technologies, USA) for 2 h at room temperature. Fluorescence images were obtained under a fluorescence microscope (Fluoview 300; Olympus, Japan).

Lee J.S., Kim E., Han S., Kang K.L, & Heo J.S. (2017). Evaluating the oxysterol combination of 22(S)-hydroxycholesterol and 20(S)-hydroxycholesterol in periodontal regeneration using periodontal ligament stem cells and alveolar bone healing models. Stem Cell Research & Therapy, 8, 276.

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Fluorescent Nanoparticle Interactions with Streptococcus pyogenes

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S. pyogenes at ~10⁹ Colony Forming Unit (CFU)/mL, was incubated without or with 10% v/v of OFNDs, OLNDs, Ery-OFNDs, Ery-OLNDs—previously labeled with 7% of fluorescein isothiocyanate (FITC, Sigma-Aldrich)—for different incubation times and in agitation at 37 °C [24 (link),32 (link)]. After 3 or 24 h of incubation, bacteria were harvested by centrifugation at 3000× g at RT for 10 min, washed 2–3× with phosphate-buffered saline (PBS 1x) to remove unbound NDs. After the final wash, each sample was resuspended in 1 mL of PBS 1x and 50 μL of all the NDs were transferred onto separate glass slides, heat-fixed, and then stained for 15 min with 5 μg/mL of a propidium iodide (Invitrogen-Thermo Fisher Scientific Inc., Waltham, MA, USA) solution in a humid chamber at 37 °C. S. pyogenes was fixed with mounting solution and covered by cover slips. All ND formulations were observed using an Olympus IX70 inverted laser scanning confocal microscope, and images were recorded by using a FluoView 300 software (Olympus Biosystems, Melville, NY, USA) [24 (link),32 (link)].

Mandras N., Luganini A., Argenziano M., Roana J., Giribaldi G., Tullio V., Cavallo L., Prato M., Cavalli R., Cuffini A.M., Allizond V, & Banche G. (2023). Design, Characterization, and Biological Activities of Erythromycin-Loaded Nanodroplets to Counteract Infected Chronic Wounds Due to Streptococcus pyogenes. International Journal of Molecular Sciences, 24(3), 1865.

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Immunofluorescence Staining of β-Catenin

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hBMSCs that undergone modulation of strategy one were fixed in 4% paraformaldehyde in PBS for 15 min, rinsed in 0.25% Triton X-100 in PBS, and subsequently blocked in 1% BSA in poly butylene succinate-co-butylene terephthalate (PBST, 0.05% Tween-20 in PBS) for 30 min. After three washes of PBS with 5 min for each time, cells were incubated for 1 h with primary anti-β-catenin antibodies at 1:100 dilution at room temperature. After the first-round incubation and three washes of PBS again, cells were incubated for 30 min with fluorescein isothiocyanate (FITC)-linked goat anti-rabbit IgG conjugated at 1:100 dilution. Finally, cell samples were washed for 40-6-Diamidino-2-phenylindole (DAPI, 1:1000 dilution, Thermo Fisher Scientific, Waltham, MA, USA) staining to highlight nuclei of the cells. Then, the cellular samples were observed by using a confocal microscope (Fluoview 300, Olympus, Tokyo, Japan), and captured images were acquired using the Image-Pro Plus 6.0 software (Media Cybernetics, Rockville, MD, USA).

Hong G., He X., Shen Y., Chen X., Yang F., Yang P., Pang F., Han X., He W, & Wei Q. (2019). Chrysosplenetin promotes osteoblastogenesis of bone marrow stromal cells via Wnt/β-catenin pathway and enhances osteogenesis in estrogen deficiency-induced bone loss. Stem Cell Research & Therapy, 10, 277.

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Broccoli Freezing Microscopy Protocol

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Broccoli samples were taken from the stalk. Three separate groups of vegetables were prepared: (1) control group, (2) fast freezing- samples frozen in liquid nitrogen, and 3) slow freezing – samples kept at −20 °C for a few days. The following steps were typical for procedures used in fluorescence microscopy. First, samples were sliced in 100–200 µm thin slices, and the preparations were stained with acridine orange and rinsed in deionized water. Then the tissue was placed on a microscope slide.
In this research, a laser scanning confocal microscope FluoView300 (Olympus Corporation, Tokyo, Japan) equipped with a UPlanSApo 10×/0.40 and UPlanSApo 20×/0.75 lens was used. Recorded images were transferred to a computer with a resolution of 2048 × 2048 or 1024 × 1024. Obtained microscopic images were processed and analysed in the Matlab R2010a (MathWorks, Natick, MA, USA.) program. The statistical calculations were performed in the Statistica calculation package (StatSoft, Inc., Tulsa, OK, USA).

Wieczorek M.N., Pieczywek P.M., Cybulska J., Zdunek A, & Jeleń H.H. (2022). Chemical Changes in the Broccoli Volatilome Depending on the Tissue Treatment. Molecules, 27(2), 500.

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Rat BMSC Scaffold Characterization

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Rat BMSCs (5 × 10⁴ cells/well) were seeded onto experimental scaffolds in 12-well plates and incubated at 37°C in a humidified atmosphere with 5% CO₂. After 1 day of culture, the samples were fixed in 2.5% glutaraldehyde and serially dehydrated with an increasing ethanol gradient, air-dried in a hood, and sputtered with gold before observation under SEM (S-3000N, Hitachi, Japan). Cytoskeletal organization was observed under a confocal laser scanning microscope (CLSM; FluoView-300, Olympus, Tokyo, Japan). Nuclei were stained with 4′,6-diamidino-2′-phenylindole (DAPI; Vector Laboratories, Burlingame, CA, USA) and actin filaments were stained with rhodamine phalloidin (Molecular Probes, Eugene, OR, USA) after culturing for 24 h. The cell spreading areas were measured using Image J software (National Institutes of Health, Bethesda, MD, USA) employing a random sampling method. Cell proliferation was assayed using a CCK-8 kit (Dojindo, Japan) at 1 day, 3 days, and 7 days of culture, with the absorbance being read at a wavelength of 450 nm, using an enzyme linked immunosorbent assay reader (Bio-Rad, Hercules, CA, USA).

Zhang X., Meng S., Huang Y., Xu M., He Y., Lin H., Han J., Chai Y., Wei Y, & Deng X. (2015). Electrospun Gelatin/β-TCP Composite Nanofibers Enhance Osteogenic Differentiation of BMSCs and In Vivo Bone Formation by Activating Ca2+-Sensing Receptor Signaling. Stem Cells International, 2015, 507154.

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Quantifying Intracellular ROS Levels

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The cells were plated on 12-well dish and confocal dishes. The cells were then incubated in the dark with 10 μM CM-H₂DCFDA for 1 h at 37°C. The cells were visualized by using laser confocal microscopy (FluoView™ 300, Olympus, Tokyo, Japan) with an excitation wavelength of 448 nm and an emission wavelength of 515 nm. To quantify the ROS production, cells were incubated with 10 μM of CM-H₂DCFDA, and washed twice with PBS. The 1 × 10⁵ cells were loaded into a 96-well black plate and assessed by using a luminometer (Victor3, Perkin-Elmer, Waltham, MA, USA).

Lee K.H., Lee S.J., Lee H.J., Choi G.E., Jung Y.H., Kim D.I., Gabr A.A., Ryu J.M, & Han H.J. (2017). Amyloid β1-42 (Aβ1-42) Induces the CDK2-Mediated Phosphorylation of Tau through the Activation of the mTORC1 Signaling Pathway While Promoting Neuronal Cell Death. Frontiers in Molecular Neuroscience, 10, 229.

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Intracellular Ca2+ Measurement using Fluo 3-AM

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The changes in [Ca²⁺]_i were observed using Fluo 3-AM dissolved in DMSO. The cells were washed once with a PBS, incubated in PBS containing 3 μM of Fluo 3-AM with 5% CO₂ at 37°C for 40 min, and washed once with the PBS and scanned every second using confocal microscopy (FluoView™ 300, Olympus, Tokyo, Japan). The fluorescence was excited at 488 nm and the emitted light was read at 515 nm. In order to verify the assay, ionomycin was applied to the cells as a positive control. All of the analysis of [Ca²⁺]_i were processed at a single cell level are expressed as the relative fluorescence intensity (RFI).

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