Mitochondria and cytoskeleton imaged on EVOS® FL Auto imaging system

BPAE cells were plated on coverslips overnight.  The next day cells were fixed and permeabilized using the Image-iT® Fixation/Permeabilization Kit (R37602) according to protocol.  Cells were then incubated with 3 µg/mL anti-ATP synthase subunit IF1 (A21355) for labeling of mitochondria for 30 min at room temperature and washed three times with dPBS and incubated with a 1:1000 dilution of Alexa Fluor® 750-labeled secondary antibody (A16079) for 30 min and washed three times in dPBS. Following staining with NucBlue® Fixed cell stain (R37606, nucleus, blue) and ReadyProbes® ActinGreen™ 488 (R37110, actin fibers, green), coverslips were mounted using ProLong® Gold antifade reagent (P36930). Images were taken using the EVOS® FL Auto Imaging System and a 10X objective (AMEP4623).

BPAE cells were plated on coverslips overnight.  The next day cells were fixed and permeabilized using the Image-iT® Fixation/Permeabilization Kit (R37602) according to protocol.  Cells were then incubated with 3 µg/mL anti-ATP synthase subunit IF1 (A21355) for labeling of mitochondria for 30 min at room temperature and washed three times with dPBS and incubated with a 1:1000 dilution of Alexa Fluor® 750-labeled secondary antibody (A16079) for 30 min and washed three times in dPBS. Following staining with NucBlue® Fixed cell stain (R37606, nucleus, blue) and ReadyProbes® ActinGreen™ 488 (R37110, actin fibers, green), coverslips were mounted using ProLong® Gold antifade reagent (P36930). Images were taken using the EVOS® FL Auto Imaging System and a 10X objective (AMEP4623).

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