Cells were fixed with 4% paraformaldehyde in PBS for 5 min at room temperature and permeabilized with 1% Triton X-100 in PBS for 5 min at room temperature. Fluorescent visualization of KDM4A was performed using pAb anti-KDM4A antibody (1:100, NB110-40585; Novus Biologicals) coupled with an Alexa Fluor 488 Goat Anti-Rabbit IgG (H+L) secondary antibody (catalogue # 4412, Cell Signaling, 1:1,000). Polymerized actin was visualized using CF™568 phalloidin (catalogue # 00044-300U; Biotium). DNA was stained with 4′,6′-diamidino-2-phenylindole (DAPI). Immunofluorescent images were captured using a × 60 lens (1.42 NA, splan APO), mounted on an FV1000 confocal laser scanning microscope driven by FluoView software (Olympus). Image acquisition was performed at room temperature, the brightness/contrast tool in FluoView software (Olympus) was used to process images and montages were made using Photoshop CS6 (Adobe).
Fluoview software
Manufactured by Olympus
Sourced in Japan, United States, Germany, Canada, Panama
Fluoview software is a digital imaging platform used for the acquisition, processing, and analysis of fluorescence microscopy data. It provides a user-friendly interface for controlling Olympus confocal microscope systems and managing image acquisition and analysis workflows.
Automatically generated - may contain errors
Lab products found in correlation
Fv1000, by Olympus (37 mentions)
Fv1000 confocal microscope, by Olympus (27 mentions)
Fv1000 confocal laser scanning microscope, by Olympus (26 mentions)
Fluoview fv1000, by Olympus (12 mentions)
Fv1000d, by Olympus (10 mentions)
Fluoview fv1000 confocal microscope, by Olympus (10 mentions)
Fluoview 1000, by Olympus (9 mentions)
Vectashield reagent, by Vector Laboratories (7 mentions)
Blocking one reagent, by Nacalai Tesque (7 mentions)
Fluoview 500 laser scanning confocal microscope, by Olympus (7 mentions)
Fv1200, by Olympus (6 mentions)
Fv1000 ix81, by Olympus (6 mentions)
Fluo 4 am, by Thermo Fisher Scientific (6 mentions)
Ix81 inverted microscope, by Olympus (5 mentions)
Bz x viewer, by Keyence (5 mentions)
Stage top incubator, by Tokai Hit (5 mentions)
Fv3000 confocal microscope, by Olympus (5 mentions)
Fv1000 confocal laser microscope, by Olympus (5 mentions)
Fv1200 ix83 laser scanning confocal microscope, by Olympus (5 mentions)
Clampfit 10, by Molecular Devices (5 mentions)
415 protocols using fluoview software
1
Immunofluorescent Visualization of KDM4A
2
Live Confocal Imaging of Zebrafish
3
Immunofluorescence Analysis of EMT Markers
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Immunofluorescence was performed with the PathScan EMT Duplex IF Kit Primary Antibody Cocktail (Cell Signaling Technology), according to the manufacturer's instructions with minor modifications 28 (link), 32 (link). Immunofluorescence analysis was performed using confocal microscopy and analyzed with the FluoView Software (Olympus Optical, Tokyo, Japan).
4
Actin Visualization Protocol
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IF was performed with a F-Actin Visualization Biochem Kit (Cytoskeleton) according to the manufacturer’s instructions and our protocol previously reported22 (link)40 41 (link). IF was observed using confocal microscopy and analyzed with the FluoView Software (Olympus Optical)22 (link)40 41 (link).
5
Confocal Imaging of Neuromuscular Synapses
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DLM neuromuscular synapse preparations were examined by immunocytochemistry essentially as described previously (Kawasaki et al., 2004 (link)). Imaging was performed using an Olympus FV1000 confocal microscope (Olympus Optical, Tokyo, Japan) with a PlanApo 60×1.4 numerical aperture oil objective (Olympus Optical) and a z-step size of 0.2 µm. The same methods were utilized to examine coxal muscle neuromuscular synapses in the leg. Images were obtained and processed with Fluoview software (Olympus Optical). Images shown are representative of those obtained from at least three different preparations.
6
Immunofluorescence Staining of Transfectants
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The transfectants were plated on chamber slides (Becton-Dickinson Falcon, Franklin Lakes, NJ, USA) at 50% confluency, washed with ice-cold PBS, fixed with 4% paraformaldehyde-PBS for 20 min, and then permeabilized in PBS containing 0.2% Triton X-100 as described previously 24 . Fixed cells were incubated with a blocking solution containing 0.5% bovine serum albumin for 1 hour at room temperature and incubated with primary antibodies overnight at 4°C. We used the following primary antibodies: rabbit anti-ARNT2 polyclonal antibody (Santa Cruz Biotechnology) and mouse anti-GLUT-1 monoclonal antibody (Arigo Biolaboratories). After washing with PBS, the cells were incubated with secondary antibodies for 1 hour at room temperature. The secondary antibodies were fluorescein isothiocyanate-conjugated anti-rabbit IgG antibody (Vector Laboratories, Burlingame, CA, USA) or Texas Red-conjugated anti-mouse IgG antibody (Vector Laboratories) incubated for 1 hour at room temperature in the dark. Finally, the sections were washed three times with PBS and mounted using Mounting Medium with DAPI (Vector Laboratories). The immunofluorescence was performed by confocal microscopy and analyzed using FluoView Software (Olympus Optical, Tokyo, Japan).
7
Visualization of exosome uptake in cells
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We visualized exosomes as reported previously17 (link). In brief, SAS, HaCaT cells and NB1RGB cells were seeded in 8–well chamber slides at a density of 2 × 104 (link) cells/well. After 24 h, the slides were washed twice in D-PBS ( −), and Endothelial Cell Media 2 containing exosomes (5 ng/µL, 10 ng/µL and 20 ng/µL) derived from NB1RGB cells (NB1RGB exo) or from oeEBI3 NB1RGB cells (EBI3 exo) stained by SYTO RNA Select (Thermo Fisher Scientific, Waltham, MA, USA) was added into each well. The exosome-treated cells were cultured for 1 h, 3 h and 6 h at 37 °C under a 5% CO2 humidified atmosphere. Then, they were treated with 4% paraformaldehyde solution at room temperature for 20 min. After staining of the nuclei using the ProLong Gold Antifade Reagent with 4′,6–diamidino-2-phenylindole (DAPI; Thermo Fisher Scientific), coverslips were added and the cells visualized under a confocal laser scanning microscope (LSM710; Carl Zeiss, Oberkochen, Germany) and analyzed by FluoView Software (Olympus Optical, Tokyo, Japan).
8
Quantifying Carotenoid and Chlorophyll in Apple Fruit
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Sections of 120, 135, and 150 D apple fruit tissue [150–250 nm cut using a VT1000S vibratome (Leica Microscopy Systems Ltd., Heerbrugg, Switzerland)] were taken from the equatorial region of each apple, mounted in 0.1 M phosphate buffer and imaged using an FV3000 laser scanning confocal (Olympus Optical Co. Ltd., Tokyo, Japan) on an IX83 inverted microscope platform (Olympus Optical Co. Ltd., Tokyo, Japan). To quantify differences in carotenoid and chlorophyll intensity between different fruit, lambda scans were performed using a 488 nm laser for excitation, collecting stepwise auto fluorescence emissions between 500 and 750 nm with a bandwidth of 10 nm and a step size 5 nm. To visualize carotenoid and chlorophyll accumulation in fruit sections, a 30-μm z-stack at 1-μm steps was collected for spectral bands between 500 and 550 nm, and 650–700 nm, respectively. Images were processed using Olympus Fluoview software to give a maximum intensity projection (Olympus Optical Co. Ltd., Tokyo, Japan).
9
Laser Scanning Confocal Microscopy Imaging
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The slides were placed on the stage of an Olympus FV1000 laser scanning confocal microscope controlled using the FluoView software. GFP and Alexa-647 were imaged using 473-nm and 647-nm lasers, respectively. The background fluorescence was measured and subtracted for each image. The fluorescence intensity was compared to the negative control slices, which did not receive any primary antibodies. Immunostained tissue was analyzed using a semiautomatic laser scanning confocal microscope (Olympus FV1000) controlled by the FluoView software. Multiple brain sections were imaged using identical microscope settings. Eighty-micrometer z-stacks were obtained from the PL region in the mPFC, and region of interest (ROI) analysis was used for quantification. The background fluorescence was measured for each image and then subtracted. The intensity quantification was performed using the FluoView Olympus software and NIH Image J.
10
Super-Resolution Confocal Microscopy with Deconvolution
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Confocal-based super-resolution microscopy was achieved with a reduced pinhole size and deconvolution (Lam et al., 2017 (link)). Super-resolution imaging was performed on an Olympus FV3000 confocal laser scanning microscope with a ×60 (1.40 NA) oil-immersion objective and high-sensitivity spectral detector using FluoView Olympus software. The Airy disk pinhole size was set to 0.4 (81 µm), and ×15 optical zoom was used. The pixel size was 26.7 nm, with an x-y optical resolution of 148.6 nm. Z-stacks were acquired with 0.2 µm steps. All images were deconvolved with Olympus cellSens software using default values. Imaris v.7.6.5 was used to generate 3D models. To account for the lower z-plane resolution, a step size of 0.1 µm was used to create the 3D models.
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