Detection of mitochondrial fission with orientation-dependent optical Fourier filters.
We utilize a recently developed optical imaging method based on Fourier processing with Gabor-like filters to detect changes in light scattering resulting from alterations in mitochondrial structure in endothelial cells undergoing apoptosis. Imaging based on Gabor filters shows a significant decrease in the orientation of subcellular organelles at 60 to 100 minutes following apoptosis induction and concomitant with mitochondrial fragmentation observed by fluorescence. The optical scatter changes can be detected at low resolution at the whole cell level. At high resolution, we combine fluorescence imaging of the mitochondria with optical Fourier-based imaging to demonstrate that the dynamic decrease in organelle orientation measured by optical Gabor filtering is spatially associated with fluorescent mitochondria and remains largely absent from nonfluorescent subcellular regions. These results provide strong evidence that the optical Gabor responses track mitochondrial fission during apoptosis and can be used to provide label-free, rapid monitoring of this morphological process within single cells.
Department of Biomedical Engineering, Rutgers University, Piscataway, NJ 08854, USA.
Robert M
RM
Pasternack
Jing-Yi
JY
Zheng
Nada N
NN
Boustany
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002417:Cattle; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000648:ultrastructure; D005374:Filtration / Q000295:instrumentation* / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D005583:Fourier Analysis; D007090:Image Interpretation, Computer-Assisted / Q000295:instrumentation / Q000379:methods; D008928:Mitochondria / Q000502:physiology* / Q000648:ultrastructure; D055095:Optics and Photonics / Q000295:instrumentation / Q000379:methods*; D015388:Organelles / Q000502:physiology / Q000648:ultrastructure; D009949:Orientation / Q000502:physiology; D012542:Scattering, Radiation; D013194:Staining and Labeling / Q000379:methods; D013347:Subcellular Fractions / Q000502:physiology
D005456:Fluorescent Dyes