Photoconversion of CFP to study neuronal tissue with electron microscopy.
Being able to use versatile light microscopy on live or fixed samples followed by electron microscopy imaging for high resolution analyses is a challenging goal. The advantage is of course that tracing and localizing fluorescently labeled molecules yields great information about dynamic cellular processes, while electron microscopy of the same sample provides exquisite information about subcellular structures. Here, I describe the straightforward combination of both methods by photoconversion of diaminobenzidine (DAB) through cyan fluorescent protein (CFP) tagged proteins localized to the Golgi apparatus in primary hippocampal neurons.
Center of Anatomy, Department Anatomy and Embryology, University Medical Center Göttingen, Kreuzbergring 36, 37075, Göttingen, Germany, Nina.wittenmayer@med.uni-goettingen.de.
Nina
N
Wittenmayer
D015100:3,3'-Diaminobenzidine / Q000737:chemistry; D000818:Animals; D002478:Cells, Cultured; D006056:Golgi Apparatus / Q000378:metabolism / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000737:chemistry*; D006624:Hippocampus / Q000166:cytology; D051379:Mice; D046529:Microscopy, Electron, Transmission; D009474:Neurons / Q000378:metabolism / Q000648:ultrastructure*; D055668:Photochemical Processes; D061251:Primary Cell Culture; D021381:Protein Transport; D017208:Rats, Wistar; D013194:Staining and Labeling
C485182:Cyan Fluorescent Protein; D049452:Green Fluorescent Proteins; D015100:3,3'-Diaminobenzidine