Name: Methoden voor in situ kwantificering van mitochondriale morfologie in spier- en terminale Schwann-cellen van muizen (Video) | JoVE Journal
Uploaded: 2026-04-10T13:00:00.000+00:00
Duration: 8 min 13 s
Description: Leer een reproduceerbare methode voor het labelen en kwantificeren van mitochondriale morfologie in intact spierweefsel en terminale Schwann-cellen van muizen met confocale microscopie.

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</ol>

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<table><tbody><tr><td>3-in-1 Whole Animal System</td><td>Aurora Scientific, Ontario, Canada</td><td>1300A</td><td>Spierfunctiesysteem met analysesoftware</td></tr><tr><td>AIVIA&nbsp;</td><td>Leica Microsystems, Bellevue, WA, USA</td><td>15</td><td>AI-gestuurde beeldanalysesoftware</td></tr><tr><td>anti-laminin&nbsp;</td><td>MilliporeSigma</td><td>L9393-.2mL</td><td>Myofiber boarder label</td></tr><tr><td>B6; D2-Tg(S100-EGFP)1Wjt/J Muisstam</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>Cryostat</td></tr><tr><td>D2.B10-DMDD2.mdx/J Muisstam</td><td>Jackson Labs, Bar Harbor, ME</td><td>JAX:13141</td><td>D2.mdx</td></tr><tr><td>DBA/2J Muisstam</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>Mitochondrial stain reconstitution reagent</td></tr><tr><td>Hoechst 33342</td><td>Thermo Fisher Scientific</td><td>H1399</td><td>Cell-permeant nucleic acid stain</td></tr><tr><td>ImageJ/Fiji&nbsp;</td><td>National Institutes of Health, Bethesda, MD, USA</td><td>SCR_002285</td><td>2D analysesoftware</td></tr><tr><td>Invitrogen ProLong Gold antifade reagent met DAPI&nbsp;</td><td>Fisher Scientific, Hampton, NJ, USA</td><td>P36941</td><td>Mounting medium</td></tr><tr><td>KimWipe</td><td>Kimberly-Clark, Irving, TX, USA</td><td>06-666A</td><td>Laboratoriumdoekjes</td></tr><tr><td>Leica LAS_X software&nbsp;</td><td>Leica Biosystems, Wetzlar, Germany</td><td>SCR_013673</td><td>Confocal software</td></tr><tr><td>MitoView Fix 640</td><td>Biotium</td><td>70075-50ug</td><td>Verrerode fluorescerende mitochondriale vlek</td></tr><tr><td>NIS-Elements Advanced Research software&nbsp;</td><td>Nikon, Melville, NY, USA</td><td>SCR_014329</td><td>Dwarsdoorsnedeanalysesoftware</td></tr><tr><td>Permount Mounting Medium&nbsp;</td><td>Fisher Scientific, Waltham, MA 02454, USA</td><td>#SP15-100</td><td>Algemeen montagemedium</td></tr><tr><td>Prism 9 software&nbsp;</td><td>GraphPad Software, La Jolla, CA, USA</td><td>RRID: SCR_002798</td><td>Statistische analysesoftware</td></tr><tr><td>Rhodamine (TRITC)</td><td>Fisher Scientific; Hampton, NJ, USA</td><td>AP132RMI</td><td>Secundaire antilichaam</td></tr><tr><td>Stellaris 5 White Light Confocal Microscope en bijbehorende deconvolutiesoftware</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>Confocal microscope</td></tr><tr><td>SZ61</td><td>Olympus, Breinigsville, PA, USA</td><td>sz61</td><td>Stereomicroscoop</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>Compound voor optimale snijtemperatuur</td></tr><tr><td>Visikol I</td><td>Sigma-Aldrich, St. Louis, MO, USA</td><td>H1-30</td><td>Weefselhelderende oplossing 1</td></tr><tr><td>Visikol II</td><td>Sigma-Aldrich, St. Louis, MO, USA</td><td>H2-30</td><td>Weefselhelderende oplossing 2</td></tr><tr><td>&alpha;-bungarotoxine 552</td><td>Biotrend, Koln, Germany</td><td>00014</td><td>Acetylcholine receptor kleurstof</td></tr></tbody></table>
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methoden voor in situ kwantificering van mitochondriale morfologie in spier- en terminale schwann-cellen van muizen

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

Methoden voor in situ kwantificering van mitochondriale morfologie in spier- en terminale Schwann-cellen van muizen

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

methods-for-situ-quantification-mitochondrial-morphology-muscle

Research

JoVE Journal

Biology

Biologie

Visualiseren individuele cellen in dicht opeengepakte weefsels, zoals terminal Schwann-cellen (SC) op neuromusculaire verbindingen (NMS), is een uitdaging. "Sequential foto-bleken" laat de afbakening van een enkele terminal SC's, bijvoorbeeld in de triangularis sterni spier explantatie, een handige zenuw-spier voorbereid, indien sequentiële bleken kan worden gecombineerd met time-lapse imaging...

Sequential Photo-bleken om Single Schwann cellen afbakenen op de neuromusculaire junctie

Skeletspieren is essentieel voor de motoriek en is het lichaam 'belangrijkste eiwit store. Muscle gezondheid metingen binnen C. elegans beschreven. Prospectieve veranderingen in de spier structuur en functie worden beoordeeld met behulp van gelokaliseerde GFP en kationische kleurstoffen.

Methoden om subcellulaire compartimenten van Muscle beoordelen C. elegans

Dit protocol maakt gebruik van driedimensionale (3D) beeldvorming en analyse technieken om te visualiseren en te kwantificeren van zenuw-specifieke mitochondriën. De technieken zijn van toepassing op andere situaties waar een fluorescent signaal wordt gebruikt voor het isoleren van een subset van gegevens uit een ander fluorescent signaal.

Driedimensionale beeldvorming en analyse van Mitochondria binnen menselijke Intraepidermal zenuwvezels

Het visualiseren en analyseren van mitochondriën uit hersenweefsel van zoogdieren is een uitdagende taak. Hier beschrijven we hoe driedimensionale (3D) reconstructie-analyse van de seriële blok-face scanning elektronenmicroscopie (SBFSEM) kan worden gebruikt om inzicht te krijgen in de morfologische en volumetrische analyse van dit cruciale energieproducerende orgaantje.

Analyse van de Brain Mitochondriën gebruik van Serial Block-Face Scanning Electron Microscopy

We presenteren een protocol voor het bestuderen van de verdeling van mitochondriën en endoplasmisch reticulum in hele cellen na genetische modificatie met behulp van correlatief licht en volume elektronenmicroscopie met inbegrip van ascorbaat peroxidase 2 en mierikswortelperoxidase kleuring, serie snijden van cellen met en zonder het doel gen in dezelfde sectie, en seriële beeldvorming via elektronenmicroscopie.

Mitochondriën en Endoplasmisch reticulum beeldvorming door Correlatief licht en volume elektronenmicroscopie

Deze studie schetst kwantitatieve metingen van synaptische grootte en lokalisatie, spier morfologie, en mitochondriale vorm in C. elegans met behulp van vrij beschikbare beeldverwerkings hulpmiddelen. Deze aanpak maakt het mogelijk toekomstige studies in C. elegans te kwantificatief vergelijken de omvang van weefsel en organel structurele veranderingen als gevolg van genetische mutaties.

Kwantitatieve benaderingen voor de studie van cellulaire structuren en organelle morfologie in Caenorhabditis elegans

Het vermogen om neuromusculaire verbindingscomponenten nauwkeurig te detecteren is cruciaal bij het evalueren van wijzigingen in de architectuur vanwege pathologische of ontwikkelingsprocessen. Hier presenteren we een volledige beschrijving van een eenvoudige methode om hoogwaardige beelden te verkrijgen van neuromusculaire verbindingen die kunnen worden gebruikt om kwantitatieve metingen uit te voeren.

Dissectie van enkelvoudige skeletspiervezels voor immunofluorescente en morfometrische analyses van neuromusculaire knooppunten van de hele berg

Dit protocol beschrijft een methode voor morfometrische analyse van neuromusculaire juncties door gecombineerde confocale en STED-microscopie die wordt gebruikt om pathologische veranderingen in muismodellen van SMA- en ColQ-gerelateerd CMS te kwantificeren.

Karakterisering van neuromusculaire juncties bij muizen door gecombineerde confocale en superresolutiemicroscopie

Dit protocol biedt een gestandaardiseerde benadering voor het in beeld brengen van de mitochondriale morfologie in meerdere weefsels van C. elegans tijdens veroudering.

Beeldvorming en kwantificering van mitochondriale morfologie in C. elegans tijdens veroudering

Het volgende protocol schetst het proces van het pancreas dissectie voor virtuele slice beeldvorming, en de daaropvolgende kwantificering van alle GFP-gelabelde beta-cellen in de gehele alvleesklier.

In situ kwantificering van beta-celmassa in Muizen

Leer een reproduceerbare methode voor het labelen en kwantificeren van mitochondriale morfologie in intact spierweefsel en terminale Schwann-cellen van muizen met confocale microscopie.

Methoden voor in situ kwantificering van mitochondriale morfologie in spier- en terminale Schwann-cellen van muizen

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