Approaches toward super-resolution fluorescence imaging of mitochondrial proteins using PALM.
Mitochondria are difficult targets for microscopy because of their small size and highly compartmentalized, membranous interior. Super-resolution fluorescence microscopy methods have recently been developed that exceed the historical limits of optical imaging. Here we outline considerations and techniques in preparing to image the relative location of mitochondrial proteins using photoactivated localization microscopy (PALM). PALM and similar methods have the capacity to dramatically increase our ability to image proteins within mitochondria, and to expand our knowledge of the location of macromolecules beyond the current limits of immunoEM.
Brown
Timothy A
TA
Howard Hughes Medical Institute, Janelia Farm Research Campus, 19700 Helix Drive, Ashburn, VA 20147-2408, USA. brownt@janelia.hhmi.org
Fetter
Richard D
RD
Tkachuk
Ariana N
AN
Clayton
David A
DA
D000818:Animals; D016616:Cryoultramicrotomy; D017110:Freeze Substitution; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D041681:NIH 3T3 Cells; D016613:Plastic Embedding; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D008164:Luminescent Proteins; D024101:Mitochondrial Proteins; D011993:Recombinant Fusion Proteins