Name: Metodi per la quantificazione in situ della morfologia mitocondriale nelle cellule muscolari e terminali di Schwann di topi (Video) | JoVE Journal
Uploaded: 2026-04-10T13:00:00.000+00:00
Duration: 8 min 13 s
Description: Scopri un metodo riproducibile per quantificare la morfologia mitocondriale tridimensionale in muscoli scheletrici e cellule di Schwann terminali di topi. Questo protocollo utilizza marcatura in vivo e microscopia confocale per analizzare l'architettura mitocondriale in tessuti neuromuscolari intatti.

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<ol><li>Memme, J. M., Slavin, M., Moradi, N., Hood, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+bioenergetics+and+turnover+during+chronic+muscle+disuse%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">Mitochondrial bioenergetics and turnover during chronic muscle disuse.</a> Int J Mol Sci. 22 (10), 5179(2021).</li>
<li>Iqbal, S., Hood, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+role+of+mitochondrial+fusion+and+fission+in+skeletal+muscle+function+and+dysfunction%5BTitle%5D)+AND+%22Front+Biosci%22%5BJournal%5D)">The role of mitochondrial fusion and fission in skeletal muscle function and dysfunction.</a> Front Biosci. 20 (1), 157-172 (2015).</li>
<li>Hyatt, H. W., Powers, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+dysfunction+is+a+common+denominator+linking+skeletal+muscle+wasting+due+to+disease%2C+aging%2C+and+prolonged+inactivity%5BTitle%5D)+AND+%22Antioxidants%22%5BJournal%5D)">Mitochondrial dysfunction is a common denominator linking skeletal muscle wasting due to disease, aging, and prolonged inactivity.</a> Antioxidants. 10 (4), 588(2021).</li>
<li>Katti, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+network+configuration+influences+sarcomere+and+myosin+filament+structure+in+striated+muscles%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Mitochondrial network configuration influences sarcomere and myosin filament structure in striated muscles.</a> Nat Commun. 13, 6058(2022).</li>
<li>Glancy, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Visualizing+mitochondrial+form+and+function+within+the+cell%5BTitle%5D)+AND+%22Trends+Mol+Med%22%5BJournal%5D)">Visualizing mitochondrial form and function within the cell.</a> Trends Mol Med. 26 (1), 58-70 (2020).</li>
<li>Yan, Z., Okutsu, M., Akhtar, Y. N., Lira, V. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Regulation+of+exercise-induced+fiber+type+transformation%2C+mitochondrial+biogenesis%2C+and+angiogenesis+in+skeletal+muscle%5BTitle%5D)+AND+%22J+Appl+Physiol%22%5BJournal%5D)">Regulation of exercise-induced fiber type transformation, mitochondrial biogenesis, and angiogenesis in skeletal muscle.</a> J Appl Physiol. 110 (1), 264-274 (2011).</li>
<li>Powers, S. K., Nelson, W. B., Hudson, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Exercise-induced+oxidative+stress+in+humans%3A+Cause+and+consequences%5BTitle%5D)+AND+%22Free+Radic+Biol+Med%22%5BJournal%5D)">Exercise-induced oxidative stress in humans: Cause and consequences.</a> Free Radic Biol Med. 51 (5), 942-950 (2011).</li>
<li>Hyatt, H., Deminice, R., Yoshihara, T., Powers, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+dysfunction+induces+muscle+atrophy+during+prolonged+inactivity%3A+A+review+of+the+causes+and+effects%5BTitle%5D)+AND+%22Arch+Biochem+Biophys%22%5BJournal%5D)">Mitochondrial dysfunction induces muscle atrophy during prolonged inactivity: A review of the causes and effects.</a> Arch Biochem Biophys. 662, 49-60 (2019).</li>
<li>Drake, J. C., Yan, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitophagy+in+maintaining+skeletal+muscle+mitochondrial+proteostasis+and+metabolic+health+with+ageing%5BTitle%5D)+AND+%22J+Physiol%22%5BJournal%5D)">Mitophagy in maintaining skeletal muscle mitochondrial proteostasis and metabolic health with ageing.</a> J Physiol. 595 (20), 6391-6399 (2017).</li>
<li>Chen, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+dysfunction%3A+Roles+in+skeletal+muscle+atrophy%5BTitle%5D)+AND+%22J+Transl+Med%22%5BJournal%5D)">Mitochondrial dysfunction: Roles in skeletal muscle atrophy.</a> J Transl Med. 21 (1), 503(2023).</li>
<li>Zhang, Z., Sliter, D. A., Bleck, C. K. E., Ding, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fis1+deficiencies+differentially+affect+mitochondrial+quality+in+skeletal+muscle%5BTitle%5D)+AND+%22Mitochondrion%22%5BJournal%5D)">Fis1 deficiencies differentially affect mitochondrial quality in skeletal muscle.</a> Mitochondrion. 49, 217-226 (2019).</li>
<li>Silva, K. A. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Angiotensin+II+suppresses+autophagy+and+disrupts+ultrastructural+morphology+and+function+of+mitochondria+in+mouse+skeletal+muscle%5BTitle%5D)+AND+%22J+Appl+Physiol%22%5BJournal%5D)">Angiotensin II suppresses autophagy and disrupts ultrastructural morphology and function of mitochondria in mouse skeletal muscle.</a> J Appl Physiol. 126 (6), 1550-1562 (2019).</li>
<li>Cattin, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Macrophage-induced+blood+vessels+guide+Schwann+cell-mediated+regeneration+of+peripheral+nerves%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">Macrophage-induced blood vessels guide Schwann cell-mediated regeneration of peripheral nerves.</a> Cell. 162 (5), 1127-1139 (2015).</li>
<li>Caillaud, M., Richard, L., Vallat, J. M., Desmoulière, A., Billet, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Peripheral+nerve+regeneration+and+intraneural+revascularization%5BTitle%5D)+AND+%22Neural+Regen+Res%22%5BJournal%5D)">Peripheral nerve regeneration and intraneural revascularization.</a> Neural Regen Res. 14 (1), 24-33 (2019).</li>
<li>Girouard, M. P., Bueno, M., Julian, V., Drake, S., Byrne, A. B., Fournier, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+molecular+interplay+between+axon+degeneration+and+regeneration%5BTitle%5D)+AND+%22Dev+Neurobiol%22%5BJournal%5D)">The molecular interplay between axon degeneration and regeneration.</a> Dev Neurobiol. 78 (10), 978-990 (2018).</li>
<li>Belanger, K., Dinis, T. M., Taourirt, S., Vidal, G., Kaplan, D. L., Egles, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Recent+strategies+in+tissue+engineering+for+guided+peripheral+nerve+regeneration%5BTitle%5D)+AND+%22Macromol+Biosci%22%5BJournal%5D)">Recent strategies in tissue engineering for guided peripheral nerve regeneration.</a> Macromol Biosci. 16 (4), 472-481 (2016).</li>
<li>Waller, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Experiments+on+the+section+of+the+glossopharyngeal+and+hypoglossal+nerves+of+the+frog%2C+and+observations+of+the+alterations+produced+thereby+in+the+structure+of+their+primitive+fibers%5BTitle%5D)+AND+%22Philos+Trans+R+Soc+Lond%22%5BJournal%5D)">Experiments on the section of the glossopharyngeal and hypoglossal nerves of the frog, and observations of the alterations produced thereby in the structure of their primitive fibers.</a> Philos Trans R Soc Lond. 140, 423-429 (1850).</li>
<li>Napoli, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+central+role+for+the+ERK-signaling+pathway+in+controlling+Schwann+cell+plasticity+and+peripheral+nerve+regeneration+in+vivo%5BTitle%5D)+AND+%22Neuron%22%5BJournal%5D)">A central role for the ERK-signaling pathway in controlling Schwann cell plasticity and peripheral nerve regeneration in vivo.</a> Neuron. 73 (4), 729-742 (2012).</li>
<li>Jessen, K. R., Mirsky, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+repair+Schwann+cell+and+its+function+in+regenerating+nerves%5BTitle%5D)+AND+%22J+Physiol%22%5BJournal%5D)">The repair Schwann cell and its function in regenerating nerves.</a> J Physiol. 594 (13), 3521-3531 (2016).</li>
<li>Hastings, R. L., Mikesh, M., Lee, Y. I., Thompson, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Morphological+remodeling+during+recovery+of+the+neuromuscular+junction+from+terminal+Schwann+cell+ablation+in+adult+mice%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Morphological remodeling during recovery of the neuromuscular junction from terminal Schwann cell ablation in adult mice.</a> Sci Rep. 10 (1), 11132(2020).</li>
<li>Glancy, B., Kim, Y., Katti, P., Willingham, T. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+functional+impact+of+mitochondrial+structure+across+subcellular+scales%5BTitle%5D)+AND+%22Front+Physiol%22%5BJournal%5D)">The functional impact of mitochondrial structure across subcellular scales.</a> Front Physiol. 11, 541040(2020).</li>
<li>Stahon, K. E., Bastian, C., Griffith, S., Kidd, G. J., Brunet, S., Baltan, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Age-related+changes+in+axonal+and+mitochondrial+ultrastructure+and+function+in+white+matter%5BTitle%5D)+AND+%22J+Neurosci%22%5BJournal%5D)">Age-related changes in axonal and mitochondrial ultrastructure and function in white matter.</a> J Neurosci. 36 (39), 9990-10001 (2016).</li>
<li>Suh, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+fragmentation+and+donut+formation+enhance+mitochondrial+secretion+to+promote+osteogenesis%5BTitle%5D)+AND+%22Cell+Metab%22%5BJournal%5D)">Mitochondrial fragmentation and donut formation enhance mitochondrial secretion to promote osteogenesis.</a> Cell Metab. 35 (2), 345-360.e7 (2023).</li>
<li>Laker, R. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+novel+MitoTimer+reporter+gene+for+mitochondrial+content%2C+structure%2C+stress%2C+and+damage+in+vivo%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">A novel MitoTimer reporter gene for mitochondrial content, structure, stress, and damage in vivo.</a> J Biol Chem. 289 (17), 12005-12015 (2014).</li>
<li>Fernando, C. A., Pangan, A. M., Cornelison, D., Segal, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Recovery+of+blood+flow+regulation+in+microvascular+resistance+networks+during+regeneration+of+mouse+gluteus+maximus+muscle%5BTitle%5D)+AND+%22J+Physiol%22%5BJournal%5D)">Recovery of blood flow regulation in microvascular resistance networks during regeneration of mouse gluteus maximus muscle.</a> J Physiol. 597 (5), 1401-1417 (2019).</li>
<li>Hammers, D. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+D2.mdx+mouse+as+a+preclinical+model+of+the+skeletal+muscle+pathology+associated+with+Duchenne+muscular+dystrophy%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">The D2.mdx mouse as a preclinical model of the skeletal muscle pathology associated with Duchenne muscular dystrophy.</a> Sci Rep. 10 (1), 14070(2020).</li>
<li>Willingham, T. B., Ajayi, P. T., Glancy, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Subcellular+specialization+of+mitochondrial+form+and+function+in+skeletal+muscle+cells%5BTitle%5D)+AND+%22Front+Cell+Dev+Biol%22%5BJournal%5D)">Subcellular specialization of mitochondrial form and function in skeletal muscle cells.</a> Front Cell Dev Biol. 9, 757305(2021).</li>
<li>Ramos, S. V., Hughes, M. C., Delfinis, L. J., Bellissimo, C. A., Perry, C. G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+bioenergetic+dysfunction+in+the+D2.mdx+model+of+Duchenne+muscular+dystrophy+is+associated+with+microtubule+disorganization+in+skeletal+muscle%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Mitochondrial bioenergetic dysfunction in the D2.mdx model of Duchenne muscular dystrophy is associated with microtubule disorganization in skeletal muscle.</a> PLoS One. 15 (10), e0237138(2020).</li>
<li>Rosen, H. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Inhibition+of+mitochondrial+fission+protein+Drp1+ameliorates+myopathy+in+the+D2-mdx+model+of+Duchenne+muscular+dystrophy%5BTitle%5D)+AND+%22bioRxiv%22%5BJournal%5D)">Inhibition of mitochondrial fission protein Drp1 ameliorates myopathy in the D2-mdx model of Duchenne muscular dystrophy.</a> bioRxiv. , (2024).</li>
<li>National Research Council. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=%22Guide+for+the+Care+and+Use+of+Laboratory+Animals%22">Guide for the Care and Use of Laboratory Animals</a>. , 8th ed, National Academies Press. Washington, DC. (2011).</li>
<li>Ino, D., Iino, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Schwann+cell+mitochondria+as+key+regulators+in+the+development+and+maintenance+of+peripheral+nerve+axons%5BTitle%5D)+AND+%22Cell+Mol+Life+Sci%22%5BJournal%5D)">Schwann cell mitochondria as key regulators in the development and maintenance of peripheral nerve axons.</a> Cell Mol Life Sci. 74 (5), 827-835 (2017).</li>
<li>Pareyson, D., Piscosquito, G., Moroni, I., Salsano, E., Zeviani, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Peripheral+neuropathy+in+mitochondrial+disorders%5BTitle%5D)+AND+%22Lancet+Neurol%22%5BJournal%5D)">Peripheral neuropathy in mitochondrial disorders.</a> Lancet Neurol. 12 (10), 1011-1024 (2013).</li>
<li>Schröder, J. M., Sommer, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+abnormalities+in+human+sural+nerves%3A+Fine+structural+evaluation+of+cases+with+mitochondrial+myopathy%2C+hereditary+and+non-hereditary+neuropathies%2C+and+review+of+the+literature%5BTitle%5D)+AND+%22Acta+Neuropathol%22%5BJournal%5D)">Mitochondrial abnormalities in human sural nerves: Fine structural evaluation of cases with mitochondrial myopathy, hereditary and non-hereditary neuropathies, and review of the literature.</a> Acta Neuropathol. 82 (6), 471-482 (1991).</li>
<li>Schröder, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neuropathy+associated+with+mitochondrial+disorders%5BTitle%5D)+AND+%22Brain+Pathol%22%5BJournal%5D)">Neuropathy associated with mitochondrial disorders.</a> Brain Pathol. 3 (2), 177-190 (1993).</li>
<li>Santosa, K. B., Keane, A. M., Jablonka-Shariff, A., Vannucci, B., Snyder-Warwick, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Clinical+relevance+of+terminal+Schwann+cells%3A+An+overlooked+component+of+the+neuromuscular+junction%5BTitle%5D)+AND+%22J+Neurosci+Res%22%5BJournal%5D)">Clinical relevance of terminal Schwann cells: An overlooked component of the neuromuscular junction.</a> J Neurosci Res. 96 (7), 1125-1135 (2018).</li>
<li>Morton, A. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Inducible+deletion+of+endothelial+cell+Efnb2+delays+capillary+regeneration+and+attenuates+myofibre+reinnervation+following+myotoxin+injury+in+mice%5BTitle%5D)+AND+%22J+Physiol%22%5BJournal%5D)">Inducible deletion of endothelial cell Efnb2 delays capillary regeneration and attenuates myofibre reinnervation following myotoxin injury in mice.</a> J Physiol. 602 (19), 4907-4927 (2024).</li>
<li>Morton, A. B., Norton, C. E., Jacobsen, N. L., Fernando, C. A., Cornelison, D. D. W., Segal, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Barium+chloride+injures+myofibers+through+calcium-induced+proteolysis+with+fragmentation+of+motor+nerves+and+microvessels%5BTitle%5D)+AND+%22Skelet+Muscle%22%5BJournal%5D)">Barium chloride injures myofibers through calcium-induced proteolysis with fragmentation of motor nerves and microvessels.</a> Skelet Muscle. 9 (1), 27(2019).</li>
<li>Sukhorukov, V. M., Dikov, D., Reichert, A. S., Meyer-Hermann, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Emergence+of+the+mitochondrial+reticulum+from+fission+and+fusion+dynamics%5BTitle%5D)+AND+%22PLoS+Comput+Biol%22%5BJournal%5D)">Emergence of the mitochondrial reticulum from fission and fusion dynamics.</a> PLoS Comput Biol. 8 (10), e1002745(2012).</li>
<li>Ferramosca, A., Zara, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+carriers+and+substrate+transport+network%3A+A+lesson+from+Saccharomyces+cerevisiae%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">Mitochondrial carriers and substrate transport network: A lesson from Saccharomyces cerevisiae.</a> Int J Mol Sci. 22 (16), (2021).</li>
</ol>

<table><tbody><tr><td>3-in-1 Whole Animal System</td><td>Aurora Scientific, Ontario, Canada</td><td>1300A</td><td>Muscle funciton system with analysis software</td></tr><tr><td>AIVIA&nbsp;</td><td>Leica Microsystems, Bellevue, WA, USA</td><td>15</td><td>Software di analisi delle immagini basato su AI</td></tr><tr><td>anti-laminin&nbsp;</td><td>MilliporeSigma</td><td>L9393-.2mL</td><td>Etichetta del bordo del miofibrill</td></tr><tr><td>B6; D2-Tg(S100-EGFP)1Wjt/J Mouse Strain</td><td>Jackson Labs, Bar Harbor, ME</td><td>JAX:005621</td><td>S100&beta;</td></tr><tr><td>Cryostar NX50 Cryostat</td><td>Epredia, Kalamazoo, MI, USA</td><td>https://www.epredia.com/products/histology-instruments/cryotomy/cryostar-nx50</td><td>Criostato</td></tr><tr><td>D2.B10-DMDD2.mdx/J Mouse Strain</td><td>Jackson Labs, Bar Harbor, ME</td><td>JAX:13141</td><td>D2.mdx</td></tr><tr><td>DBA/2J Mouse Strain</td><td>Jackson Labs, Bar Harbor, ME</td><td>JAX:00671</td><td>WT</td></tr><tr><td>Dimethyl Sulfoxide (DMSO)</td><td>Fisher Scientific; Hampton, NJ, USA</td><td>BP231-100</td><td>Reagente di ricostituzione per colorazione mitocondriale</td></tr><tr><td>Hoechst 33342</td><td>Thermo Fisher Scientific</td><td>H1399</td><td>Colorante per acido nucleico permeabile alle cellule</td></tr><tr><td>ImageJ/Fiji&nbsp;</td><td>National Institutes of Health, Bethesda, MD, USA</td><td>SCR_002285</td><td>Software di analisi 2D</td></tr><tr><td>Invitrogen ProLong Gold antifade reagent with DAPI&nbsp;</td><td>Fisher Scientific, Hampton, NJ, USA</td><td>P36941</td><td>Medium di montaggio</td></tr><tr><td>KimWipe</td><td>Kimberly-Clark, Irving, TX, USA</td><td>06-666A</td><td>Salviettine da laboratorio</td></tr><tr><td>Leica LAS_X software&nbsp;</td><td>Leica Biosystems, Wetzlar, Germany</td><td>SCR_013673</td><td>Software confocale</td></tr><tr><td>MitoView Fix 640</td><td>Biotium</td><td>70075-50ug</td><td>Colorante mitocondriale fluorescente a infrarossi</td></tr><tr><td>NIS-Elements Advanced Research software&nbsp;</td><td>Nikon, Melville, NY, USA</td><td>SCR_014329</td><td>Software di analisi dell'area della sezione trasversale</td></tr><tr><td>Permount Mounting Medium&nbsp;</td><td>Fisher Scientific, Waltham, MA 02454, USA</td><td>#SP15-100</td><td>Medium di montaggio generico</td></tr><tr><td>Prism 9 software&nbsp;</td><td>GraphPad Software, La Jolla, CA, USA</td><td>RRID: SCR_002798</td><td>Software di analisi statistica</td></tr><tr><td>Rhodamine (TRITC)</td><td>Fisher Scientific; Hampton, NJ, USA</td><td>AP132RMI</td><td>Anticorpo secondario</td></tr><tr><td>Stellaris 5 White Light Confocal Microscope and corresponding deconvolution software</td><td>Leica Microsystems, Wetzlar, Germany</td><td>158301313:158201310:158004752: 158301312:158301314:158401100: 158209011:158301130:158204510: 158301120:158401140:158204511: 15500332:15506428:15506517:155 13859:15521522:15525226:155252 32:15525314:15555009:15555017: 15555046:27100022:155933660: 155933666:155933668:158000640 :158002407:158003150:158004141 :158004201:158004421:158004423 :158200681:158202140:158203114: 158203200:158203220:158204201: 158204510:158204709:158301200: 9I_LL_STELLARIS_C:9T-CLSM-APP _CLASSC:9WE_LAS_405:9WE_ LAS_WLL:9WE_LL_STELLARIS_C</td><td>Microscopio confocale</td></tr><tr><td>SZ61</td><td>Olympus, Breinigsville, PA, USA</td><td>sz61</td><td>Microscopio stereo</td></tr><tr><td>Tissue-Plus O.C.T. Compound&nbsp;</td><td>Scigen, Fischer Scientific, Hampton, NJ, USA</td><td>23-730-571</td><td>Composto per temperatura di taglio ottimale</td></tr><tr><td>Visikol I</td><td>Sigma-Aldrich, St. Louis, MO, USA</td><td>H1-30</td><td>Soluzione per la decolorazione dei tessuti 1</td></tr><tr><td>Visikol II</td><td>Sigma-Aldrich, St. Louis, MO, USA</td><td>H2-30</td><td>Soluzione per la decolorazione dei tessuti 2</td></tr><tr><td>&alpha;-bungarotoxin 552</td><td>Biotrend, Koln, Germany</td><td>00014</td><td>Colorante per il recettore dell'acetilcolina</td></tr></tbody></table>

metodi per la quantificazione in situ della morfologia mitocondriale nelle cellule muscolari e terminali di schwann di topi

Methods for In situ Quantification of Mitochondrial Morphology in Muscle and Terminal Schwann Cells of Mice

Metodi per la quantificazione in situ della morfologia mitocondriale nelle cellule muscolari e terminali di Schwann di topi

Mitochondrial network morphology is widely used as an indicator of cellular health; however, quantifying mitochondrial architecture within intact tissues ...

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Biologia

Visualizzare le singole celle di densamente tessuti confezionati, come le cellule di Schwann terminali (SC) a giunzioni neuromuscolari (NMJs), è una sfida. "Sequenziale foto-bleaching" permette di delineare SC singolo terminale, per esempio nel triangularis espianto muscolo Sterni, un comodo nervo-muscolo preparazione, dove sequenziale sbiancamento può essere combinato con time-lapse imaging...

Sequential Photo-sbiancante per Delineare cellule di Schwann singoli sulla giunzione neuromuscolare

Il muscolo scheletrico è essenziale per la locomozione ed è il principale negozio di proteine ​​dei corpi. Muscle misure sanitarie all'interno C. elegans sono descritti. I potenziali modifiche alla struttura e la funzione muscolare, valutati in base GFP localizzate e coloranti cationici.

Metodi di valutazione subcellulari Compartimenti di muscoli in C. elegans

Questo protocollo utilizza tecniche di imaging e l'analisi tridimensionali (3D) per visualizzare e quantificare i mitocondri del nervo specifico. Le tecniche sono applicabili ad altre situazioni in cui un segnale fluorescente viene utilizzato per isolare un sottoinsieme di dati da un altro segnale fluorescente.

Imaging tridimensionale e l'analisi dei mitocondri all'interno di fibre nervose umane Intraepidermal

La visualizzazione e l'analisi mitocondriale dal tessuto cerebrale dei mammiferi è un compito impegnativo. Qui, descriviamo come l'analisi di ricostruzione tridimensionale (3D) dalla microscopia elettronica a scansione seriale a blocchi (SBFSEM) possa essere utilizzata per ottenere informazioni sull'analisi morfologica e volumetrica di questo organello critico generatore di energia.

Analisi di Brain mitocondri Utilizzando Serial Block-Face Scanning Electron Microscopy

Presentiamo un protocollo per studiare la distribuzione di mitocondri e reticolo endoplasmico in intere cellule dopo la modificazione genetica utilizzando la microscopia elettronica di luce e volume correlata, tra cui l'ascorbate perossidasi 2 e la colorazione della perossidia di rafano, sezionamento seriale di cellule con e senza il gene bersaglio nella stessa sezione e imaging seriale tramite microscopia elettronica.

Mitocondri e reticulum imaging endoplasmico di microscopia elettronica di luce correlata e volume

Questo studio descrive le misurazioni quantitative delle dimensioni sinaptiche e della localizzazione, della morfologia muscolare e della forma mitocondriale in C. elegans utilizzando strumenti di elaborazione delle immagini liberamente disponibili. Questo approccio consente a studi futuri in C. elegans di confrontare quantitativamente l'estensione dei cambiamenti strutturali dei tessuti e degli organelli a seguito di mutazioni genetiche.

Approcci quantitativi per studiare le strutture cellulari e la morfologia delle orgine in Caenorhabditis elegans

La capacità di rilevare con precisione i componenti della giunzione neuromuscolare è fondamentale per valutare le modifiche nella sua architettura a causa di processi patologici o di sviluppo. Qui presentiamo una descrizione completa di un metodo semplice per ottenere immagini di alta qualità di giunzioni neuromuscolari a montaggio intero che possono essere utilizzate per eseguire misurazioni quantitative.

Dissezione di fibre muscolari scheletriche singole per analisi immunofluorescenti e morfometriche di giunzioni neuromuscolari a montaggio intero

Questo protocollo descrive un metodo per l'analisi morfometrica delle giunzioni neuromuscolari mediante microscopia combinata confocale e STED che viene utilizzata per quantificare i cambiamenti patologici in modelli murini di SMA e CMS correlati a ColQ.

Caratterizzazione delle giunzioni neuromuscolari nei topi mediante microscopia combinata confocale e super-risoluzione

Questo protocollo fornisce un approccio standardizzato per l'imaging della morfologia mitocondriale in più tessuti di C. elegans durante l'invecchiamento .

Imaging e quantificazione della morfologia mitocondriale in C. elegans durante l'invecchiamento

Il protocollo seguente delinea il processo di dissezione del pancreas per l'imaging fetta virtuale, e la successiva quantificazione di tutte le GFP-tagged beta-cellule del pancreas intero.

Quantificazione in situ di cellule beta pancreatiche Messa in topi

Scopri un metodo riproducibile per quantificare la morfologia mitocondriale tridimensionale in muscoli scheletrici e cellule di Schwann terminali di topi. Questo protocollo utilizza marcatura in vivo e microscopia confocale per analizzare l'architettura mitocondriale in tessuti neuromuscolari intatti.

Metodi per la quantificazione in situ della morfologia mitocondriale nelle cellule muscolari e terminali di Schwann di topi

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