To address this problem the standard variations of the dIF assay for ALDH1 were used to isolate the r/o cells. The ALDH1 selective inhibitor (680109958 clone, Aldagen, USA; 10 m or 100 m final dilution containing 1 × 106 cells) was added to ACF-15 cells (10 m). At least 500 cells and 200 control cells were counted. The cells were fixed in 0.08% formaldehyde-H2O2 for 1 h at room temperature, washed 3 × 5 min with PBS, and then incubated for 30 min in PBS containing 1% BSA and a PE-conjugated ALDH1A1 antibody (11,840 clone, R&D systems. The cells were washed 3 × 5 min with PBS containing 1% BSA and then incubated for 30 min in PBS containing 1% BSA and a FITC-conjugated secondary antibody (11,840 clone, R&D systems; 1,000-fold diluted). After washing again with PBS containing 1% BSA, the cover slips were mounted onto slides. Confocal images were acquired with a Laser scanning confocal microscope (FV300, Olympus, Tokyo, Japan) including the subunits of Olympus Fluoview software. The images were displayed and the counts were performed with the FV10-ASW viewer software for Windows XP (Olympus).
FASTcap is a program that helps with Digital Image Acquisition (DIA) using Olympus Stream, cellSens, Olympus analySIS, Olympus CX3, Olympus XC-A10, Olympus C700, Olympus CX5/C6/C7, Olympus C-3000, Olympus C-4000, Olympus C-5000 and other Olympus Industrial Microscopes (OIM) connected to a PC with Windows XP, Vista, Windows 7 (SP1), Windows 8 or Windows 10.
Dispose of any old OLYMPUS Stream. This unit is obsolete with the introduction of the OLYMPUS Stream 5-M. We strongly recommend that you replace this OLYMPUS Stream with a new model OLYMPUS Stream, one that is upgraded to the Bluetooth capability. 7211a4ac4a