Localization of anti-TFAM antibody in live cells stained with SYBG Gold.

Live HeLa cells were transfected with anti-TFAM antibody conjugated to PF555 dye and then stained for 30 min. with SYBR Gold (final dilution 1:10000) and Mitotracker Deep Red™ (final concentration 250 nM). Images (z-stacks) of live cells were acquired on Zeiss LSM780 microscope with 63x/1.4 Oil objective; channels were acquired sequentially; detection ranges were adjusted to minimize spectral bleed-through: (500–550 nm for SYBR Gold, 565–598 nm for PF555 and 645–700 nm for Mitotracker Deep Red. Deconvolution of datasets was performed. Single optical slice from a representative field of view is shown; scale bar 10 μm. Cyan squares on the left panels mark the region of interest which is shown at a higher magnification in the right panels. (PDF)