Correction of image instability in confocal microscopy using image realignment. Effects on the analysis of intracellular calcium.
Using confocal microscopy, we have examined the increases in [Ca(2+)](i) evoked by sodium channel toxins in cells labelled with the fluorescent dye INDO-1. We describe a new image analysis method that improves the detection of region-specific, toxin-induced patterns of change of intracellular calcium. This method is based on correction of global image motion followed by calculation of the strength of correlation between calcium changes in "seed" or reference pixels chosen to represent different regions of cells and those in other regions of the image. When the selected "seed" pixel was chosen to be in either varicosities or neurites, correlations were detected in the same regions of other cells as well as in the soma, indicating specific but spatially distinct patterns of behaviour. Control images (without changes in [Ca(2+)](i)) did not reveal significant interpixel correlations. The ability to recognize correlated patterns of calcium change in different regions of cells was greatly improved by correction for global motion.
Massensini
Andre R
AR
Department of Biostatistics and Computing, Brain Image Analysis Unit, Institute of Psychiatry, King's College, De Crespigny Park, Denmark Hill, P.O. Box 22, London SE5 8AF, UK.
Romano-Silva
Marco A
MA
Suckling
John
J
Gomez
Marcus V
MV
Brammer
Michael J
MJ
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D006825:Hybridomas / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods*; D009498:Neurotoxins / Q000494:pharmacology; D012604:Scorpion Venoms / Q000494:pharmacology; D013997:Time Factors
D009498:Neurotoxins; D012604:Scorpion Venoms; C119038:Ts-gamma neurotoxin; C003609:tityustoxin; D002118:Calcium