Xingguo Liu, Liang Yang, Qi Long, David Weaver, György Hajnóczky. Choosing proper fluorescent dyes, proteins, and imaging techniques to study mitochondrial dynamics in mammalian cells. Biophysics Reports, 2017, 3(4-6): 64-72. doi: 10.1007/s41048-017-0037-8
Citation: Xingguo Liu, Liang Yang, Qi Long, David Weaver, György Hajnóczky. Choosing proper fluorescent dyes, proteins, and imaging techniques to study mitochondrial dynamics in mammalian cells. Biophysics Reports, 2017, 3(4-6): 64-72. doi: 10.1007/s41048-017-0037-8

Choosing proper fluorescent dyes, proteins, and imaging techniques to study mitochondrial dynamics in mammalian cells

doi: 10.1007/s41048-017-0037-8
Funds:  This work was financially supported by the "973" program (2013CB967403, 2012CB721105, and 2016YFA0100302), the "Frontier Science Key Research Program" of the Chinese Academy of Sciences (QYZDB-SSW-SMC001), the National Natural Science Foundation projects of China (31622037, 31271527, 81570520, 31601176, and 31601088), Guangzhou Science and Technology Program (2014Y2-00161), Guangzhou Health Care and Cooperative Innovation Major Project (201604020009), Guangdong Natural Science Foundation for Distinguished Young Scientists (S20120011368), Guangdong Province Science and Technology Innovation The Leading Talents Program (2015TX01R047), Guangdong Province Science and Technology Innovation Young Talents Program (2014TQ01R559), Guangdong Province Science and Technology Program (2015A020212031), the PhD Start-up Fund of Natural Science Foundation of Guangdong Province (2014A030310071), and by NIH Grants (AA017773 and DK051526) to G.H.
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  • Corresponding author: Xingguo Liu, György Hajnóczky
  • Received Date: 25 April 2016
  • Rev Recd Date: 05 September 2016
  • Publish Date: 30 November 2017
  • Mitochondrial dynamics refers to the processes maintaining mitochondrial homeostasis, including mitochondrial fission, fusion, transport, biogenesis, and mitophagy. Mitochondrial dynamics is essential for maintaining the metabolic function of mitochondria as well as their regulatory roles in cell signaling. In this review, we summarize the recently developed imaging techniques for studying mitochondrial dynamics including:mitochondrial-targeted fluorescent proteins and dyes, live-cell imaging using photoactivation, photoswitching and cell fusion, mitochondrial transcription and replication imaging by in situ hybridization, and imaging mitochondrial dynamics by super-resolution microscopy. Moreover, we discuss examples of how to choose and combine proper fluorescent dyes and/or proteins.
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  • Andersen E, Asuri N, Clay M, Halloran M (2010) Live imaging of cell motility and actin cytoskeleton of individual neurons and neural crest cells in zebrafish embryos. J Vis Exp 36:1726
    Ando R, Hama H, Yamamoto-Hino M, Mizuno H, Miyawaki A (2002) An optical marker based on the UV-induced green-tored photoconversion of a fluorescent protein. Proc Natl Acad Sci USA 99:12651-12656
    Appelhans T, Richter CP, Wilkens V, Hess ST, Piehler J, Busch KB (2012) Nanoscale organization of mitochondrial microcompartments revealed by combining tracking and localization microscopy. Nano Lett 12:610-616
    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
    Carvalho PH, Correa JR, Guido BC, Gatto CC, De Oliveira HC, Soares TA, Neto BA (2014) Designed benzothiadiazole fluorophores for selective mitochondrial imaging and dynamics. Chemistry 20:15360-15374
    Chang H, Zhang M, Ji W, Chen J, Zhang Y, Liu B, Lu J, Zhang J, Xu P, Xu T (2012) A unique series of reversibly switchable fluorescent proteins with beneficial properties for various applications. Proc Natl Acad Sci USA 109:4455-4460
    Chatre L, Ricchetti M (2015) mTRIP:an imaging tool to investigate mitochondrial DNA dynamics in physiology and disease at the single-cell resolution. Methods Mol Biol 1264:133-147
    Chazotte B (2011) Labeling mitochondria with MitoTracker dyes. Cold Spring Harb Protoc 2011:990-992
    Chen H, Detmer SA, Ewald AJ, Griffin EE, Fraser SE, Chan DC (2003) Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development. J Cell Biol 160:189-200
    Chudakov DM, Belousov VV, Zaraisky AG, Novoselov VV, Staroverov DB, Zorov DB, Lukyanov S, Lukyanov KA (2003) Kindling fluorescent proteins for precise in vivo photolabeling. Nat Biotechnol 21:191-194
    Cornaglia M, Mouchiroud L, Marette A, Narasimhan S, Lehnert T, Jovaisaite V, Auwerx J, Gijs MA (2015) An automated microfluidic platform for C. elegans embryo arraying, phenotyping, and long-term live imaging. Sci Rep 5:10192
    Detmer SA, Chan DC (2007) Functions and dysfunctions of mitochondrial dynamics. Nat Rev Mol Cell Biol 8:870-879
    Ekanayake SB, El Zawily AM, Paszkiewicz G, Rolland A, Logan DC (2015) Imaging and analysis of mitochondrial dynamics in living cells. Methods Mol Biol 1305:223-240
    Fernandez-Suarez M, Ting AY (2008) Fluorescent probes for super-resolution imaging in living cells. Nat Rev Mol Cell Biol 9:929-943
    Friedman JR, Nunnari J (2014) Mitochondrial form and function. Nature 505:335-343
    Fukasawa Y, Tsuji J, Fu SC, Tomii K, Horton P, Imai K (2015) MitoFates:improved prediction of mitochondrial targeting sequences and their cleavage sites. Mol Cell Proteom 14:1113-1126
    Gustafsson MG (2000) Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. J Microsc 198:82-87
    Gustafsson MG (2005) Nonlinear structured-illumination microscopy:wide-field fluorescence imaging with theoretically unlimited resolution. Proc Natl Acad Sci USA 102:13081-13086
    Hell SW (2003) Toward fluorescence nanoscopy. Nat Biotechnol 21:1347-1355
    Huang LJ, Wang L, Ma Y, Durick K, Perkins G, Deerinck TJ, Ellisman MH, Taylor SS (1999) NH2-Terminal targeting motifs direct dual specificity A-kinase-anchoring protein 1 (D-AKAP1) to either mitochondria or endoplasmic reticulum. J Cell Biol 145:951-959
    Huang S, Han R, Zhuang Q, Du L, Jia H, Liu Y, Liu Y (2015a) New photostable naphthalimide-based fluorescent probe for mitochondrial imaging and tracking. Biosens Bioelectron 71:313-321
    Huang S, Han R, Zhuang Q, Du L, Jia H, Liu Y, Liu Y (2015b) New photostable naphthalimide-based fluorescent probe for mitochondrial imaging and tracking. Biosens Bioelectron 71:313-321
    Jakobs S, Wurm CA (2014) Super-resolution microscopy of mitochondria. Curr Opin Chem Biol 20:9-15
    Jans DC, Wurm CA, Riedel D, Wenzel D, Stagge F, Deckers M, Rehling P, Jakobs S (2013) STED super-resolution microscopy reveals an array of MINOS clusters along human mitochondria. Proc Natl Acad Sci USA 110:8936-8941
    Joshi DC, Bakowska JC (2011) Determination of mitochondrial membrane potential and reactive oxygen species in live rat cortical neurons. J Vis Exp 51:e2704
    Karbowski M, Cleland MM, Roelofs BA (2014) Photoactivatable green fluorescent protein-based visualization and quantification of mitochondrial fusion and mitochondrial network complexity in living cells. Methods Enzymol 547:57-73
    Katajisto P, Dohla J, Chaffer CL, Pentinmikko N, Marjanovic N, Iqbal S, Zoncu R, Chen W, Weinberg RA, Sabatini DM (2015) Stem cells. Asymmetric apportioning of aged mitochondria between daughter cells is required for stemness. Science 348:340-343
    Klein T, Loschberger A, Proppert S, Wolter S, van de Linde S, Sauer M (2011) Live-cell dSTORM with SNAP-tag fusion proteins. Nat Methods 8:7-9
    Kukat C, Wurm CA, Spahr H, Falkenberg M, Larsson NG, Jakobs S (2011) Super-resolution microscopy reveals that mammalian mitochondrial nucleoids have a uniform size and frequently contain a single copy of mtDNA. Proc Natl Acad Sci USA 108:13534-13539
    Kuzmenko A, Tankov S, English BP, Tarassov I, Tenson T, Kamenski P, Elf J, Hauryliuk V (2011) Single molecule tracking fluorescence microscopy in mitochondria reveals highly dynamic but confined movement of Tom40. Sci Rep 1:195
    Liesa M, Shirihai OS (2013) Mitochondrial dynamics in the regulation of nutrient utilization and energy expenditure. Cell Metab 17:491-506
    Liu X, Weaver D, Shirihai O, Hajnoczky G (2009) Mitochondrial ‘kiss-and-run’:interplay between mitochondrial motility and fusion-fission dynamics. EMBO J 28:3074-3089
    Logan DC, Leaver CJ (2000) Mitochondria-targeted GFP highlights the heterogeneity of mitochondrial shape, size and movement within living plant cells. J Exp Bot 51:865-871
    Lovy A, Molina AJ, Cerqueira FM, Trudeau K, Shirihai OS (2012) A faster, high resolution, mtPA-GFP-based mitochondrial fusion assay acquiring kinetic data of multiple cells in parallel using confocal microscopy. J Vis Exp 65:e3991
    Lukinavicius G, Umezawa K, Olivier N, Honigmann A, Yang G, Plass T, Mueller V, Reymond L, Correa IR Jr, Luo ZG, Schultz C, Lemke EA, Heppenstall P, Eggeling C, Manley S, Johnsson K (2013) A near-infrared fluorophore for live-cell superresolution microscopy of cellular proteins. Nat Chem 5:132-139
    Malka F, Guillery O, Cifuentes-Diaz C, Guillou E, Belenguer P, Lombes A, Rojo M (2005) Separate fusion of outer and inner mitochondrial membranes. EMBO Rep 6:853-859
    Martelli PL, Savojardo C, Fariselli P, Tasco G, Casadio R (2015) Computer-based prediction of mitochondria-targeting peptides. Methods Mol Biol 1264:305-320
    Matsushima R, Hamamura Y, Higashiyama T, Arimura S, Sodmergen Tsutsumi N, Sakamoto W (2008) Mitochondrial dynamics in plant male gametophyte visualized by fluorescent live imaging. Plant Cell Physiol 49:1074-1083
    McBride HM, Neuspiel M, Wasiak S (2006) Mitochondria:more than just a powerhouse. Curr Biol:CB 16:R551-R560
    Mitra K, Lippincott-Schwartz J (2010) Analysis of mitochondrial dynamics and functions using imaging approaches. Curr Protoc Cell Biol Chapter 4:Unit 4 25 21-21
    Owens GC, Edelman DB (2015) Photoconvertible fluorescent protein-based live imaging of mitochondrial fusion. Methods Mol Biol 1313:237-246
    Owens GC, Walcott EC (2012) Extensive fusion of mitochondria in spinal cord motor neurons. PLoS ONE 7:e38435
    Patterson GH, Lippincott-Schwartz J (2002) A photoactivatable GFP for selective photolabeling of proteins and cells. Science 297:1873-1877
    Patterson G, Davidson M, Manley S, Lippincott-Schwartz J (2010) Superresolution imaging using single-molecule localization. Ann Rev Phys Chem 61:345-367
    Pham AH, McCaffery JM, Chan DC (2012) Mouse lines with photoactivatable mitochondria to study mitochondrial dynamics. Genesis 50:833-843
    Rocha S, De Keersmaecker H, Uji-i H, Hofkens J, Mizuno H (2014) Photoswitchable fluorescent proteins for superresolution fluorescence microscopy circumventing the diffraction limit of light. Methods Mol Biol 1076:793-812
    Rust MJ, Bates M, Zhuang X (2006) Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat Methods 3:793-795
    Scaduto RC Jr, Grotyohann LW (1999) Measurement of mitochondrial membrane potential using fluorescent rhodamine derivatives. Biophys J 76:469-477
    Shcherbakova DM, Sengupta P, Lippincott-Schwartz J, Verkhusha VV (2014) Photocontrollable fluorescent proteins for superresolution imaging. Ann Rev Biophys 43:303-329
    Shim SH, Xia C, Zhong G, Babcock HP, Vaughan JC, Huang B, Wang X, Xu C, Bi GQ, Zhuang X (2012) Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes. Proc Natl Acad Sci USA 109:13978-13983
    Subach FV, Patterson GH, Manley S, Gillette JM, LippincottSchwartz J, Verkhusha VV (2009) Photoactivatable mCherry for high-resolution two-color fluorescence microscopy. Nat Methods 6:153-159
    Subach OM, Patterson GH, Ting LM, Wang Y, Condeelis JS, Verkhusha VV (2011) A photoswitchable orange-to-far-red fluorescent protein, PSmOrange. Nat Methods 8:771-777
    Suzuki H, Maeda M, Mihara K (2002) Characterization of rat TOM70 as a receptor of the preprotein translocase of the mitochondrial outer membrane. J Cell Sci 115:1895-1905
    Tatsuta T, Langer T (2008) Quality control of mitochondria:protection against neurodegeneration and ageing. EMBO J 27:306-314
    Twig G, Elorza A, Molina AJ, Mohamed H, Wikstrom JD, Walzer G, Stiles L, Haigh SE, Katz S, Las G, Alroy J, Wu M, Py BF, Yuan J, Deeney JT, Corkey BE, Shirihai OS (2008) Fission and selective fusion govern mitochondrial segregation and elimination by autophagy. EMBO J 27:433-446
    Wiedenmann J, Ivanchenko S, Oswald F, Schmitt F, Rocker C, Salih A, Spindler KD, Nienhaus GU (2004) EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion. Proc Natl Acad Sci USA 101:15905-15910
    Wilkens V, Kohl W, Busch K (2013) Restricted diffusion of OXPHOS complexes in dynamic mitochondria delays their exchange between cristae and engenders a transitory mosaic distribution. J Cell Sci 126:103-116
    Yamakoshi H, Palonpon A, Dodo K, Ando J, Kawata S, Fujita K, Sodeoka M (2015) A sensitive and specific Raman probe based on bisarylbutadiyne for live cell imaging of mitochondria. Bioorg Med Chem Lett 25:664-667
    Yang L, Long Q, Liu J, Tang H, Li Y, Bao F, Qin D, Pei D, Liu X (2015) Mitochondrial fusion provides an ‘initial metabolic complementation’ controlled by mtDNA. Cell Mol Life Sci 72:2585
    Youle RJ, van der Bliek AM (2012) Mitochondrial fission, fusion, and stress. Science 337:1062-1065
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