Mouse MI or MII oocytes were fixed in 4%(w/v) paraformaldehyde for 40 min at room temperature and washed three times (10 min each) in washing buffer (PBS containing 0.01%Triton X‐100 and 0.1% Tween‐20). The oocytes were then permeated in 1% Triton X‐100/PBS at room temperature for 1 h and washed three times (10 min each) in washing buffer. The oocytes were then blocked with blocking buffer (1% BSA /PHEM with 100 mM glycine) for 1 h at 37°C. The oocytes were incubated at 4°C overnight with anti‐α‐tubulin antibody (1:8000) diluted in blocking buffer. After washing three times with washing buffer for 10 min each, the oocytes were incubated at 37°C for 1 h with goat anti‐mouse‐FITC antibody (1:100 dilutions, CW Biotech) and washed four times in washing buffer for 10 min each. Finally, DNA was stained with 4’6‐diamidino‐2‐phenylindole (DAPI, Vector Laboratories Inc.). The oocytes were then expanded on glass slides and examined with confocal laser‐scanning microscopy (FLUOVIEW FV1000, Olympus) using the FLUOVIEW Viewer (Olympus). The excitation lasers were set at 488 nm, and emission channels of 520 nm were used for green fluorescence detection.
Fluoview viewer
Manufactured by Olympus
Sourced in Japan
The Fluoview Viewer is a software application developed by Olympus for viewing and analyzing fluorescence microscopy data. It provides a user-friendly interface for managing and visualizing fluorescence images and image sequences.
Automatically generated - may contain errors
Lab products found in correlation
Fluoview fv1000, by Olympus (3 mentions)
Fv1000, by Olympus (2 mentions)
Confocal laser scanning microscope, by Olympus (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions)
Lasersharp 2000, by Bio-Rad (1 mentions)
Fluoview fv10i, by Olympus (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Dojindo Laboratories (1 mentions)
Alexa fluor 647 labeled goat anti mouse igg, by Abcam (1 mentions)
Inverted scope, by Olympus (1 mentions)
Celltiter glo assay, by Promega (1 mentions)
Paraformaldehyde (pfa), by Thermo Fisher Scientific (1 mentions)
Click it tunel alexa fluor imaging assay, by Thermo Fisher Scientific (1 mentions)
Fv1000 confocal microscope, by Olympus (1 mentions)
Observer z1, by Zeiss (1 mentions)
Prolong diamond, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride dapi, by Thermo Fisher Scientific (1 mentions)
Fluoview fv1000 confocal microscope, by Olympus (1 mentions)
Fv1000 fluoview camera, by Olympus (1 mentions)
Saponin, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
21 protocols using fluoview viewer
1
Immunofluorescence Analysis of Mouse Oocytes
2
Confocal Fluorescence Imaging Protocol
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Confocal fluorescence images were acquired on an Olympus scanning laser microscope Fluoview FV1000 using 4X and 10X air objectives or 20X and 40X oil immersion objectives. Image analysis was performed using either ImageJ (NIH) or Fluoview Viewer version 4.2 (Olympus). Cells were counted blindly.
3
Confocal Fluorescence Microscopy Imaging
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Confocal fluorescence microscopy images were acquired on a Bio-Rad (Zeiss) Radiance 2100 Rainbow laser scanning confocal microscope with LaserSharp 2000 software (Bio-Rad, Hercules, CA), or an Olympus FV1000 laser scanning confocal microscope with Fluoview Viewer software (Olympus, Center Valley, PA), both equipped with 60× oil immersion objectives. Images were processed with Fiji software.67 (link)
4
Visualizing HP1-γ in MEF Cells
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MEF cells were plated in 12-well plates with coverglass for imaging. The MEF cells were fixed in 4% PFA and permeabilized in 5% Triton X-100–PBS. The cells were incubated with HP1-γ polyclonal antibody antibodies (1 : 250, Abcam-ab213167) overnight at 4° C. After washing twice, cells were incubated with the Alexa-conjugated secondary antibodies for 1 hour. The cells were also counterstained with Hoechst dye solution before mounting for the nuclei. The slides were observed in a laser confocal microscope at 600x magnification. Olympus Fluoview Viewer was used to image and quantify the fluorescence.
5
Confocal Fluorescence Imaging Protocol
6
Subcellular Localization of Nanoparticles
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NPC1 or normal cells were seeded on 35-mm glass-bottomed dishes at a density of 1 × 104 cells/dish and incubated overnight. After the medium was exchanged with fresh DMEM (900 μL), the treatment solutions (100 μL) were applied to the dish. After the cells were incubated with FITC-labeled HP-β-CD and HE-SS-PRX for 24 h, the cells were washed twice with PBS, fixed with 4% paraformaldehyde for 15 min at room temperature, and permeabilized with 0.1% Triton X-100 for 10 min at room temperature. Then, the cells were treated with mouse anti-human early endosome antigen 1 (EEA1) antibody (clone: 14/EEA1) (BD Biosciences) (1:100 dilution), mouse anti-human CD63 antibody (clone: H5C6) (BioLegend, San Diego, CA, USA) (1:200 dilution), or mouse anti-human lysosomal-associated membrane protein 1 (LAMP1) antibody (clone: H4A3) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) (1:50 dilution) for 1 h at room temperature. After three washes with PBS, the cells were stained with Alexa Fluor 647-labeled goat anti-mouse IgG (Abcam, Cambridge, MA, USA) (1:1000 dilution) for 30 min at room temperature. Then, the cells were stained with 4′,6-diamidino-2-phenylindole (DAPI) (Dojindo Laboratories) (1 μg/mL) for 10 min at room temperature. The CLSM observations were performed on a FluoView FV10i and the colocalization percentage was analyzed using FluoView Viewer (Olympus).
7
Quantifying Macrophage oxLDL Uptake
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Dil-labeled oxLDL was used to trace the oxLDL uptake. Mice bone marrow-derived macrophages were treated with oxLDL (50 μg/ml) for 24 h, and then washed twice with cold PBS before staining. Cold acid washing buffer (0.5 M glacial acetic acid, 150 mM sodium chloride, pH 2.5) was used to wash surface adherent ox-LDL. Cells were then fixed with 4% PFA. The cells were visualized using a confocal laser scanning microscope (Olympus, Tokyo, Japan). The mean fluorescent intensity was measured by software Olympus Fluoview Viewer. Alternatively, the cells were detached from the plate, and cell surface markers CD11b and F4/80 were stained on ice for 30 min. After two times of wash by FACS washing buffer (2% FBS in PBS), samples were analyzed by FACS. Data were further quantified by software FlowJo.
8
Multimodal Imaging Techniques
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Phase contrast, RFP and β-galactosidase staining images were captured using an Olympus inverted scope and digitally captured using Spot Advanced software NHBE (Diagnostics Instruments, Macomb, Michigan). Confocal images were captured on an Olympus Fluoview 500 scope (Olympus, Center Valley, PA) and captured using Fluoview Viewer.
9
Analyzing Mycobacterium tuberculosis-Induced Cell Death
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TUNEL staining: Transfected MDMs on coverslips were infected with mCherry M.tb at MOI 5 and 50 for 24 and 48 h. Cells were then fixed with 4% PFA (Affymetrix, Santa Clara, CA) and labeled using the Click-iT TUNEL Alexa Fluor Imaging Assay (Invitrogen) following the manufacturer’s instructions. Cells were imaged with an Olympus FV1000 confocal microscope (Olympus, Shinjuku, Japan). Using Olympus Fluoview Viewer, at least 100 MDMs were manually counted to quantify % MDMs that stained with TUNEL.
CellTiter Glo Assays: Transfected or inhibitor treated MDMs in 96 well plates were infected with M.tb at MOI 5 for 24 h and cell death was assayed in triplicate with the CellTiter Glo Assay (Promega) following the manufacturer’s instructions.
CellTiter Glo Assays: Transfected or inhibitor treated MDMs in 96 well plates were infected with M.tb at MOI 5 for 24 h and cell death was assayed in triplicate with the CellTiter Glo Assay (Promega) following the manufacturer’s instructions.
10
Immunofluorescence Staining of Cells
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