For osteogenic and adipogenic protein expression, MSCs were cultured on coverslips with ODM and ADM for 14 days. At day 7, 10, and 14, cells were fixed with 4% paraformaldehyde in PBS, followed by washing with PBS twice. Cells were permeabilized with 0.3% Triton X-100 (Merck KGaA) in PBS, non-specifically blocked with 3% BSA (Thermo Fisher Scientific, Inc.) in PBS, and incubated with mouse anti-RUNX2 antibodies and mouse anti-PPARγ antibodies (1:50; Santa Cruz Biotechnology, CA, United States) at 4°C overnight. After three washes with PBST, the cells were incubated with goat anti-mouse IgG (1:500; H+L; Alexa Fluor 488; Abcam) for 1 h at room temperature and then counterstained with antifade containing DAPI (Invitrogen; Thermo Fisher Scientific, Inc.). The fluorescent images were obtained with a confocal laser scanning microscope and analyzed with FluoView FV1000 Software version 3.01 (Olympus Corp., Tokyo, Japan).
Fluoview fv1000 software version 3
Manufactured by Olympus
Sourced in Japan
The FluoView FV1000 Software version 3.01 is a software package developed by Olympus for use with their FluoView FV1000 confocal microscope system. The software provides the core functionality for operating and configuring the microscope hardware, as well as image acquisition and data analysis capabilities.
Automatically generated - may contain errors
2 protocols using fluoview fv1000 software version 3
1
Osteogenic and Adipogenic Protein Expression in MSCs
2
Immunofluorescence Microscopy Imaging
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) × 100 Absorbance of control-Absorbance of blank Samples were then washed twice with PBS to remove the excess primary antibodies, followed by incubation with Alexa flour 488 conjugated secondary antibody at room temperature for 1 hour. Cells were washed with PBS and mounted with Antifade Mounting with DAPI solution (Invitrogen; Thermo Fisher Scientific, Inc.,). The observation was performed using a confocal laser scanning microscope, the fluorescent micrographs were captured and analyzed with FluoView FV1000 Software version 3.01 (Olympus Corp., Tokyo, Japan).
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