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Correlative cryo-fluorescence and cryo-electron microscopy (cryo-CLEM) system has been fast becoming a powerful technique with the advantage to allow the fluorescent labeling and direct visualization of the close-to-physiologic ultrastructure in cells at the same time, offering unique insights into the ultrastructure with specific cellular function. There have been various engineered ways to achieve cryo-CLEM including the commercial FEI iCorr system that integrates fluorescence microscope into the column of transmission electron microscope. In this study, we applied the approach of the cryo-CLEM-based iCorr to image the syntaphilin-immobilized neuronal mitochondria in situ to test the performance of the FEI iCorr system and determine its correlation accuracy. Our study revealed the various morphologies of syntaphilin-immobilized neuronal mitochondria that interact with microtubules and suggested that the cryo-CLEM procedure by the FEI iCorr system is suitable with a half micron-meter correlation accuracy to study the cellular organelles that have a discrete distribution and large size, e.g. mitochondrion, Golgi complex, lysosome, etc.
Abbe E, (1873) Beitrage zur theorie des mikroskops und der mikroskopischen wahrnehmung.Arch Mikr Anat 9:413-468
Al-Amoudi A, Chang JJ, Leforestier A, McDowall A, Salamin LM, Norlen LP, Richter K, Blanc NS, Studer D, Dubochet J, (2004) Cryo-electron microscopy of vitreous sections.The EMBO journal 23:3583-3588
Betzig E, Patterson GH, Sougrat R, Lindwasser OW, Olenych S, Bonifacino JS, Davidson MW, Lippincott-Schwartz J, Hess HF, (2006) Imaging intracellular fluorescent proteins at nanometer resolution.Science 313:1642-1645
Boldogh IR, Pon LA, (2007) Mitochondria on the move.Trends Cell Biol 17:502-510
Dubochet J, Adrian M, Chang JJ, Homo JC, Lepault J, McDowall AW, Schultz P, (1988) Cryo-electron microscopy of vitrified specimens.Q Rev Biophys 21:129-228
Goldstein LS, Yang Z, (2000) Microtubule-based transport systems in neurons: the roles of kinesins and dyneins.Annu Rev Neurosci 23:39-71
Gurskaya NG, Verkhusha VV, Shcheglov AS, Staroverov DB, Chepurnykh TV, Fradkov AF, Lukyanov S, Lukyanov KA, (2006) Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light.Nat Biotechnol 24:461-465
Gustafsson MG, (2000) Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy.J Microsc 198:82-87
Hanein D, Volkmann N, (2011) Correlative light-electron microscopy.Adv Protein Chem Struct Biol 82:91-99
Hell SW, Wichmann J, (1994) Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy.Opt Lett 19:780-782
Hess ST, Girirajan TP, Mason MD, (2006) Ultra-high resolution imaging by fluorescence photoactivation localization microscopy.Biophys J 91:4258-4272
Hollenbeck PJ, Saxton WM, (2005) The axonal transport of mitochondria.J Cell Sci 118:5411-5419
Jiang M, Chen G, (2006) High Ca2+-phosphate transfection efficiency in low-density neuronal cultures.Nat Protoc 1:695-700
Jun S, Ke D, Debiec K, Zhao G, Meng X, Ambrose Z, Gibson GA, Watkins SC, Zhang P, (2011) Direct visualization of HIV-1 with correlative live-cell microscopy and cryo-electron tomography.Structure 19:1573-1581
Kang JS, Tian JH, Pan PY, Zald P, Li C, Deng C, Sheng ZH, (2008) Docking of axonal mitochondria by syntaphilin controls their mobility and affects short-term facilitation.Cell 132:137-148
Klar TA, Engel E, Hell SW, (2001) Breaking Abbe’s diffraction resolution limit in fluorescence microscopy with stimulated emission depletion beams of various shapes.Phys Rev E 64:066613
Kukulski W, Schorb M, Welsch S, Picco A, Kaksonen M, Briggs JA, (2011) Correlated fluorescence and 3D electron microscopy with high sensitivity and spatial precision.J Cell Biol 192:111-119
Li D, Shao L, Chen BC, Zhang X, Zhang M, Moses B, DE Milkie , Beach JR, Hammer JA3rd, Pasham M, Kirchhausen T, Baird MA, Davidson MW, Xu P, Betzig E, (2015) Extended-resolution structured illumination imaging of endocytic and cytoskeletal dynamics.Science 349:aab3500
Lucic V, Kossel AH, Yang T, Bonhoeffer T, Baumeister W, Sartori A, (2007) Multiscale imaging of neurons grown in culture: from light microscopy to cryo-electron tomography.J Struct Biol 160:146-156
Mironov AA, Beznoussenko GV, (2009) Correlative microscopy: a potent tool for the study of rare or unique cellular and tissue events.J Microsc 235:308-321
Misgeld T, Kerschensteiner M, Bareyre FM, Burgess RW, Lichtman JW, (2007) Imaging axonal transport of mitochondria in vivo.Nat Methods 4:559-561
Nangaku M, Sato-Yoshitake R, Okada Y, Noda Y, Takemura R, Yamazaki H, Hirokawa N, (1994) KIF1B, a novel microtubule plus end-directed monomeric motor protein for transport of mitochondria.Cell 79:1209-1220
Nicholls DG, Budd SL, (2000) Mitochondria and neuronal survival.Physiol Rev 80:315-360
Nogales E, Scheres SH, (2015) Cryo-EM: a unique tool for the visualization of macromolecular complexity.Mol Cell 58:677-689
Rintoul GL, Filiano AJ, Brocard JB, Kress GJ, Reynolds IJ, (2003) Glutamate decreases mitochondrial size and movement in primary forebrain neurons.J Neurosci 23:7881-7888
Rust MJ, Bates M, Zhuang X, (2006) Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).Nat Methods 3:793-795
Sartori A, Gatz R, Beck F, Rigort A, Baumeister W, Plitzko JM, (2007) Correlative microscopy: bridging the gap between fluorescence light microscopy and cryo-electron tomography.J Struct Biol 160:135-145
Schneider CA, Rasband WS, Eliceiri KW, (2012) NIH image to imageJ: 25 years of image analysis.Nat Methods 9:671-675
Schorb M, Gaechter L, Avinoam O, Sieckmann F, Clarke M, Bebeacua C, Bykov YS, Sonnen AF, Lihl R, Briggs JA, (2016) New hardware and workflows for semi-automated correlative cryo-fluorescence and cryo-electron microscopy/tomography.J Struct Biol
Schwartz CL, Sarbash VI, Ataullakhanov FI, Mcintosh JR, Nicastro D, (2007) Cryo-fluorescence microscopy facilitates correlations between light and cryo-electron microscopy and reduces the rate of photobleaching.J Microsc-Oxford 227:98-109
Wolff G, Hagen C, Grunewald K, Kaufmann R, (2016) Towards correlative super-resolution fluorescence and electron cryo-microscopy.Biol cell
Zhang P, (2013) Correlative cryo-electron tomography and optical microscopy of cells.Curr Opin Struct Biol 23:763-770
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