Fluoview fv1000 inverted confocal microscope

Manufactured by Olympus

Sourced in Japan

The Fluoview FV1000 is an inverted confocal microscope designed for advanced fluorescence imaging. It features a high-resolution optical system and a modular design for adaptability. The microscope is capable of performing confocal laser scanning microscopy to enable detailed observation and analysis of biological samples.

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Lab products found in correlation

Uplsapo, by Olympus (1 mentions) Anti cre, by Novus Biologicals (1 mentions) Ab5392, by Abcam (1 mentions) Sc 7523, by Santa Cruz Biotechnology (1 mentions) Propidium iodide solution, by Merck Group (1 mentions) Uplfln 40 oil objective, by Olympus (1 mentions) Fluosave reagent, by Merck Group (1 mentions) Biotin labeled anti rat igg, by Jackson ImmunoResearch (1 mentions) Pe labeled streptavidin, by BioLegend (1 mentions) Pe labeled streptavidin, by Jackson ImmunoResearch (1 mentions) Cm1950, by Leica (1 mentions)

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FluoView FV1000 (1553 citations) FV1000 confocal microscope (1548 citations) FluoView FV1000 confocal microscope (570 citations) FluoView confocal microscope (117 citations) Fluoview FV1000 microscope (109 citations) FV1000 inverted confocal microscope (41 citations) Inverted confocal microscope (28 citations) FluoView FV1000 confocal (23 citations) FV1000 inverted microscope (14 citations) Confocal FV1000 (10 citations)

7 protocols using fluoview fv1000 inverted confocal microscope

Multimodal Microscopy Imaging of Synaptic Proteins

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Anti-kalirin-7 (Kal-7, rabbit polyclonal, a generous gift from R Mains, UConn Health, JH2958^{85 (link)}, 1:200), anti-Cre (rabbit polyclonal, Novus Biologicals, Centennial, CO, 1:1000) or anti-NMDAR1 (rabbit monoclonal, ab274377, Abcam, Cambridge, MA, 1:100) were used. General: All antibodies were diluted in the blocking solution and incubated with tissue sections overnight at 4 °C. After washing, sections were incubated with the appropriate fluorescent secondary antibody (Alexa fluor 488 and 594, 1:1000) for 2 h, washed in PBS 3× for 10 min, after which they were mounted on slides with ProLong Glass antifade reagent in some cases with 5% nuclear blue. Images were taken with an Olympus FluoView TM FV1000 confocal inverted microscope with objective UPLSAPO 40× or 100× NA:1:30 (Olympus, Tokyo, Japan). For the immunohistochemical analysis, the merged z-stack image (2 μm steps) was used. Image segmentation was first performed using a thresholding sub-routine in ImageJ so that the original color image was converted to a binary image. This allowed for visualization of the regions of interest (ROI) in cases where the background intensity was non-homogeneous. ROIs were then analyzed for image luminosity in the original image using Adobe Photoshop after subtracting the adjacent background levels, and the results were verified by ImageJ. 3 ROIs were analyzed per mouse.

Evrard M.R., Li M., Shen H, & Smith S.S. (2021). Preventing adolescent synaptic pruning in mouse prelimbic cortex via local knockdown of α4βδ GABAA receptors increases anxiety response in adulthood. Scientific Reports, 11, 21059.

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Immunohistochemistry of Hippocampal Proteins

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Following anesthesia with urethane (0.1 ml 40%), mice were perfused with saline (12–15 mls/min) and then with 4% paraformaldehyde followed by post-fixation of brain in 4% PFA (48 h, 4oC). The hippocampus was sectioned at 40 μm ( (Leica VT 100M vibratome), and sections washed in PBS for (10 min, (3x) followed by blocking for 2 h in 10% donkey serum ( 0.4% Triton, .05% sodium azide in phosphate buffered saline). For immunohistochemistry, either anti-Kal-7 (ab52012, Abcam, 1:200) or anti-α4 (sc7523, Santa Cruz, 1:20) and, in some cases, anti-MAP2 (microtubule-associated protein, ab5392, Abcam, 1:1000) diluted in the blocking solution overnight at 4°C were used. The fluorescent secondary antibody, either (Alexa fluor 488 or 594 (1:500) was applied for 2 h, and washed in PBS for 10 min (3x). Images were analyzed using FIJI (Image J) software after acquisition with Olympus FluoView TM FV1000 confocal inverted microscope Olympus, Tokyo, Japan.

Parato J., Shen H, & Smith S.S. (2018). α4βδ GABAA receptors trigger synaptic pruning and reduce dendritic length of female mouse CA3 hippocampal pyramidal cells at puberty. Neuroscience, 398, 23-36.

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Mitochondrial oxidative stress in neurons

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Primary neurons were plated at 1.0 × 10⁶ on 35 mm glass bottom culture PDL coated dishes (MatTek). On DIV 7, neurons were transfected with CellLight mitochondria GFP (Invitrogen) for 16-18 h. On DIV 8, neurons were incubated with 5 μM MitoPY1 (30 (link)-32 ) for 40 min in 1X HBSS medium (136.9 mM NaCl, 5 mM KCl, 20 nM HEPES, 5.5mM glucose, 0.59 mM KH₂PO₄, 0.56 mM Na₂HPO₄, 0.9 mM MgSO₄ and 1.4 mM CaCl₂) at 37 °C. Either vehicle (DMSO) or 10 nM rotenone was added at time zero; the signal was allowed to stabilize for ∼100,000 ms, after which, the data were normalized to this baseline. Regions of interest (ROI) with only CellLight positive mitochondria were analyzed. Fluorescence intensities (excitation 555nm and emission 625nm) and images were captured with a 60X lens using an Olympus Fluoview FV1000 inverted confocal microscope.

Sanders L.H., McCoy J., Hu X., Mastroberardino P.G., Dickinson B.C., Chang C.J., Chu C.T., Van Houten B, & Timothy Greenamyre J. (2014). Mitochondrial DNA damage: molecular marker of vulnerable nigral neurons in Parkinson's disease. Neurobiology of disease, 70, 214-223.

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Imaging Coculture Biofilms on Transwell

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To image coculture biofilms on transwell filters, biofilm assays were performed as described above with S. aureus USA100 expressing GFP. At the conclusion of the assay, CF AECs on transwell filters were washed once, stained with Hoechst stain (1:1,000 in MEM) for 10 min, and then fixed in cold 4% paraformaldehyde (PFA; diluted in phosphate-buffered saline [PBS]) overnight at 4°C. For live/dead staining of CF AECs during S. aureus and virus coinfections, filters were stained with propidium iodide solution (Sigma-Aldrich) diluted 1:8,000 in MEM for 15 min prior to washing and fixing. After fixing, PFA was removed and filters were washed once with cold PBS. Transwell filters were cut out from inserts, mounted on slides in ProLong Gold antifade reagent, sealed with coverslips, allowed to cure at room temperature for 24 h in the dark, and then stored at 4°C. Microscopy of fixed filters was performed on an Olympus FluoView FV1000 inverted confocal microscope. Bacterial biomass was quantified with Nikon Elements software.

Kiedrowski M.R., Gaston J.R., Kocak B.R., Coburn S.L., Lee S., Pilewski J.M., Myerburg M.M, & Bomberger J.M. (2018). Staphylococcus aureus Biofilm Growth on Cystic Fibrosis Airway Epithelial Cells Is Enhanced during Respiratory Syncytial Virus Coinfection. mSphere, 3(4), e00341-18.

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Imaging Neonatal Fibroblasts with TIVA

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CCD-1112Sk human neonatal foreskin-derived fibroblasts (ATCC, Manassas, VA) were cultured in IMDM with 10% FBS and penicillin/streptomycin in a 37 °C incubator with 5% CO₂. For microscopy experiments split cells were seeded onto P35GC-1.5-14-C 35 mm Petri dishes with a 14 mm glass microwell (MatTek Corporation, Ashland, MA) and cultured for an additional 24 h to reach 70–90% confluency. At T₀ the dishes were washed twice with DPBS and replaced with fresh serum-free media containing 0.5 μM PS-22/9/9 or 22/9/9 TIVA and then returned to the incubator. At 1.5, 6, and 24 h the dishes were removed and imaged using a Fluoview FV1000 inverted confocal microscope (Olympus) with a 1.30 NA UPLFLN 40× oil objective (Olympus), and 543 nm green HeNe laser. Images were collected for both Cy3 emission (555 nm – 625 nm bandpass filter) and Cy5 FRET emission (650 nm long-pass filter).

Yeldell S.B., Ruble B.K, & Dmochowski I.J. (2017). Oligonucleotide modifications enhance probe stability for single cell transcriptome in vivo analysis (TIVA). Organic & biomolecular chemistry, 15(47), 10001-10009.

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Single-cell Analysis of GPCR Complexes

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For single-cell analysis of 2PK-3 cells expressing Myc-tagged β₂ARs together with Flag-tagged CCR7, CXCR4, or S1PR1, cells were stained with Alexa Fluor 647–conjugated anti-Myc (9B11; Cell Signaling Technology) and biotin-labeled antibodies against CCR7 (4B12) or CXCR4 (2B11), followed by PE-labeled streptavidin (BioLegend). S1PR1 was detected with a purified rat antibody (713412; R&D Systems), followed by biotin-labeled anti–rat IgG (Jackson ImmunoResearch Laboratories) and PE-labeled streptavidin (Arnon et al., 2011 (link)). Labeled cells were seeded onto slides and fixed with 4% paraformaldehyde. Slides were mounted with FluoSave Reagent (EMD Millipore). Images were acquired at room temperature with Fluoview FV10-ASW version 3.00 using a Fluoview FV1000 inverted confocal microscope (Olympus), equipped with plan apochromat UPLSAPO objectives (100× oil immersion with 1.40 NA). Co-localization of fluorescent signals was analyzed with a plug-in ‘RG2B Colocalization’ for ImageJ software.

Nakai A., Hayano Y., Furuta F., Noda M, & Suzuki K. (2014). Control of lymphocyte egress from lymph nodes through β2-adrenergic receptors. The Journal of Experimental Medicine, 211(13), 2583-2598.

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Fluorescent Skin Permeation Imaging

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The vesicular formulations were labelled with a lipophilic fluorescent marker (Rho-PE; 0.035 mg/ml), loaded with a hydrophilic fluorescent marker (CF; 0.025 mg/ml) and applied on pig skin using Franz diffusion cells, as previously reported [31] . At regular time intervals (2, 4, 8, 24 h), skin specimens were gently washed with distilled water, the diffusion area punched out and frozen at -80 °C. Sections of the skin (7 µm thickness) were cut with a cryostat (Leica CM1950, Barcelona, Spain) orthogonally (in the z axis) to the surface, and examined under a FluoView FV1000 inverted confocal microscope (Olympus, Barcelona, Spain) equipped with an Ultraviolet/Visible light laser. Using an UplanSApo 20x objective NA 0.75, images with a field size of 1024×1024 µm were generated. CF and Rho-PE were excited at 600 nm and 559 nm, and detected at 640 nm and 578 nm, respectively.

Castangia I., Manca M.L., Caddeo C., Maxia A., Murgia S., Pons R., Demurtas D., Pando D., Falconieri D., Peris J.E., Fadda A.M., Manconi M, & Manconi M. (2015). Faceted phospholipid vesicles tailored for the delivery of Santolina insularis essential oil to the skin. Colloids and surfaces. B, Biointerfaces, 132.

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