Morphological filtering was applied as a final image processing step in order to eliminate background elements around the primary coronary arteries.
Filtering was applied to exclude spots for which the reference, Cy5, signal was less than two times the background value of that particular spot.
No further filtering was applied.
No image filtering was applied.
Filtering was applied to exclude spots with flags; for estimating the correction factor in normalization, only spots were included with Cy3 values larger than two times background.
Images were first filtered with a 1.0 pixel radius Gaussian filter to eliminate speckle, then a 100 pixel radius rolling-ball background subtract filter was applied to minimize intensity variations.
Single channels were extracted from the confocal images either for nuclei (DAPI) or for TUNEL (TMR-Red), and after application of a threshold that eliminates background dust, a watershed filter was applied on the binary images.
Single channels were extracted from the confocal images either for nuclei (DAPI) or TUNEL (TMR-Red) and after application of a threshold that eliminates background dust, a watershed filter was applied on the binary images.
Background was subtracted and in some cases a median filter was applied only for presentation.
In the 3D-constructor module of Image-Pro Plus, a 3×3×3 lo-pass 3D-filter was applied to each cropped nuclei in all channels to reduce background noise.
