HeLa cells transfected with pEGFP-N1 or pEGFP-N1-LRRC25 adhered to 24-well slides. After 48 h, the cells were fixed with 2% PFA, permeabilized with 0.5% Triton X-100, and stained with Hoechst 33342 (Sigma, USA). Stained cells were visualized using an Olympus FV10i fluorescence confocal microscope (Olympus, USA). NB4 cells treated with 1 μmol/L ATRA for 4 days were cytospun on slide glasses, fixed, and incubated with mouse anti-LRRC25ec monoclonal antibody (No. 70) or mouse IgG1 isotype control antibody (Sigma, Cat. No. M7894) for 1 h. Then, the cells were washed and incubated with FITC-conjugated goat anti-mouse IgG for 30 min. The cells were washed with PBS, permeabilized with 0.5% Triton X-100, and stained with Hoechst 33342. Photograph of stained cells were taken by confocal microscopy as described above.
Fv10i fluorescence confocal microscope
Manufactured by Olympus
Sourced in United States, Japan
The FV10i fluorescence confocal microscope is a compact, integrated imaging system designed for high-resolution, high-contrast imaging of fluorescent samples. It features a laser-scanning confocal architecture that provides optical sectioning capability for three-dimensional imaging. The FV10i is capable of capturing images with a resolution of up to 2048 x 2048 pixels.
Automatically generated - may contain errors
Lab products found in correlation
Image it fx signal enhancer, by Thermo Fisher Scientific (2 mentions)
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions)
Fluoview software, by Olympus (2 mentions)
Dapi counterstain, by Thermo Fisher Scientific (2 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Vectashield mounting medium h 1000, by Vector Laboratories (1 mentions)
Mitotracker deep red, by Thermo Fisher Scientific (1 mentions)
4 protocols using fv10i fluorescence confocal microscope
1
Visualizing LRRC25 Localization in Transfected Cells
2
Immunofluorescence Analysis of Lymph Node Tissue
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Lymph nodes were fixed in 10% formalin and dehydrated in 30% sucrose solution for 24 h at 4 °C. The tissue samples were then embedded in O.C.T. compound. Sections (8 μm) were washed with PBS-T (PBS containing 0.5% Tween-20) and permeabilized with permeabilization buffer (PBS containing 0.5% Triton X-100) for 30 min at room temperature. Then, the sections were blocked with blocking solution (PBS-T containing 5% bovine serum albumin) for 30 min at room temperature. After blocking, the sections were labeled with antibodies against rat anti-Foxp3 (17-5773-82, 1:100, eBioscience) for 1 h at room temperature. Counterstaining was performed with DAPI (cat. D9542, Sigma-Aldrich) for 10 min at 37 °C. The lymph nodes were coverslipped with Vectashield mounting medium (H-1000; Vector). Immunofluorescence images were acquired using an FV10i fluorescence confocal microscope (Olympus, Tokyo, Japan).
3
Immunofluorescence Staining of Adherent Cells
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The cells were adhered to 10-well slides, fixed, and permeabilized as previously described (21 (link)). The cells were blocked with Image-iT FX Signal Enhancer (Life Technologies, Grand Island, NY, USA) for 15 min at room temperature, and then either singly or doubly stained with primary antibodies. Subsequent labeling with Alexa Fluor-conjugated secondary antibodies and DAPI counterstain (Life Technologies) were performed to visualize primary antibodies and nuclei, respectively. Stained cells were viewed using the Olympus FV10i fluorescence confocal microscope. Images were analyzed using FluoView software (Olympus, Melville, NY, USA).
4
MitoTracker Deep Red Staining Procedure
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Live cells were incubated with 100 nM of MitoTracker Deep Red (Life Technologies, Grand Island, NY, USA) for 20 min under regular culture condition or left unstained as a negative control. Stained cells were washed with PBS, adhered to 10-well slides, fixed, and permeabilized as previously described [40 (link)]. Cells were blocked with Image-iT FX signal enhancer (Life Technologies) for 15 min at room temperature, and then either singly or doubly stained with primary antibodies. Subsequent labeling with Alexa Fluor-conjugated secondary antibodies and DAPI counterstain (Life Technologies) were performed to visualize primary antibodies and nuclei, respectively. Stained cells were viewed using the Olympus FV10i fluorescence confocal microscope. Images were analyzed using the Fluoview software (Olympus, Melville, NY, USA).
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