Name: Métodos para Quantificação In Situ da Morfologia Mitocondrial em Células Musculares e Terminais de Schwann de Camundongos (Vídeo) | JoVE Journal
Uploaded: 2026-04-10T13:00:00.000+00:00
Duration: 8 min 13 s
Description: Aprenda a quantificar a morfologia mitocondrial in situ em músculo esquelético e células de Schwann terminais de camundongos usando microscopia confocal tridimensional. Este método permite analisar redes mitocondriais em tecidos intactos sem necessidade de repórteres transgênicos.

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</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>AI driven image analysis software</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 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>Cryostat</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>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 analysis software</td></tr><tr><td>Invitrogen ProLong Gold antifade reagent with 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>Laboratory wipes</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>Far-red fluorescent mitochondrial stain</td></tr><tr><td>NIS-Elements Advanced Research software&nbsp;</td><td>Nikon, Melville, NY, USA</td><td>SCR_014329</td><td>Cross sectional area analysis software</td></tr><tr><td>Permount Mounting Medium&nbsp;</td><td>Fisher Scientific, Waltham, MA 02454, USA</td><td>#SP15-100</td><td>General mounting media</td></tr><tr><td>Prism 9 software&nbsp;</td><td>GraphPad Software, La Jolla, CA, USA</td><td>RRID: SCR_002798</td><td>Statistical analysis software</td></tr><tr><td>Rhodamine (TRITC)</td><td>Fisher Scientific; Hampton, NJ, USA</td><td>AP132RMI</td><td>Secondary antibody</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>Confocal microscope</td></tr><tr><td>SZ61</td><td>Olympus, Breinigsville, PA, USA</td><td>sz61</td><td>Stereo microscope</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>Optimal cutting temperature compound</td></tr><tr><td>Visikol I</td><td>Sigma-Aldrich, St. Louis, MO, USA</td><td>H1-30</td><td>Tissue clearing solution 1</td></tr><tr><td>Visikol II</td><td>Sigma-Aldrich, St. Louis, MO, USA</td><td>H2-30</td><td>Tissue clearing solution 2</td></tr><tr><td>&alpha;-bungarotoxin 552</td><td>Biotrend, Koln, Germany</td><td>00014</td><td>Acetylcholine receptor dye</td></tr></tbody></table>

métodos para quantificação in situ da morfologia mitocondrial em células musculares e terminais de schwann de camundongos

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

Métodos para Quantificação In Situ da Morfologia Mitocondrial em Células Musculares e Terminais de Schwann de Camundongos

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

Biologia

Visualizando as células individuais em tecidos densamente compactados, tais como as células de Schwann terminais (SCs) em junções neuromusculares (NMJs), é um desafio. "Sequential foto-branqueamento" permite delinear SCs terminais individuais, por exemplo, no explante triangularis músculo Sterni, uma preparação conveniente nervo-músculo, onde sequencial de branqueamento podem ser combinados com o lapso de tempo de imagem...

Sequencial de foto-branqueamento de delinear células de Schwann individuais na junção neuromuscular

O músculo esquelético é essencial para a locomoção e é a principal loja de proteína dos corpos. Medidas de saúde musculares dentro de C. elegans são descritos. Mudanças potenciais na estrutura e função muscular são avaliados usando GFP localizada e corantes catiônicos.

Métodos para avaliar Subcelulares Compartimentos de músculo em C. elegans

Este protocolo usa tridimensional (3D) de imagem e análise técnicas para visualizar e quantificar mitocôndrias nervo específico. As técnicas são aplicáveis a outras situações onde um sinal fluorescente é usado para isolar um subconjunto de dados de outro sinal fluorescente.

Imagem tridimensional e análise de mitocôndrias dentro as fibras nervosas intraepidérmicas humana

A visualização e análise mitocondrial do tecido cerebral de mamíferos é uma tarefa desafiadora. Aqui, descrevemos como a análise de reconstrução tridimensional (3D) da microscopia eletrônica de varredura serial (SBFSEM) pode ser usada para obter insights sobre a análise morfológica e volumétrica dessa organela geradora de energia crítica.

Análise do Cérebro mitocôndrias Usando Serial Block-cara Scanning Electron Microscopy

Nós apresentamos um protocolo para estudar a distribuição das mitocôndria e do retículo endoplasmático em pilhas inteiras após a modificação genética usando a luz correlativa e a microscopia de elétron do volume que incluem a peroxidase 2 do ascorbato e a mancha do peroxidase do rábano, seccionamento serial de células com e sem o gene alvo na mesma seção, e imagens seriadas via microscopia eletrônica.

Mitocôndria e Reticulum endoplasmático por microscopia eletrônica de luz e volume correlativo

Este estudo esboça medidas quantitativas do tamanho e da localização sináptica, da morfologia do músculo, e da forma mitochondrial em C. elegans usando ferramentas livremente disponíveis do processamento de imagem. Esta aproximação permite que os estudos futuros em C. elegans comparem quantitativamente a extensão de mudanças estruturais do tecido e do organela em conseqüência das mutações genéticas.

Abordagens quantitativas para o estudo de estruturas celulares e morfologia organelle em Caenorhabditis elegans

A capacidade de detectar com precisão componentes de junção neuromuscular é crucial na avaliação de modificações em sua arquitetura devido a processos patológicos ou de desenvolvimento. Aqui apresentamos uma descrição completa de um método simples para obter imagens de alta qualidade de junções neuromusculares de montagem inteira que podem ser usadas para realizar medições quantitativas.

Dissecção de fibras musculares esqueléticas únicas para análises imunofluorescentes e morfométricas de junções neuromusculares de monte inteiro

Este protocolo descreve um método para análise morfométrica de junções neuromusculares por microscopia confocal combinada e STED que é usado para quantificar alterações patológicas em modelos de camundongos de SMA e CMS relacionada ao ColQ.

Caracterização de Junções Neuromusculares em Camundongos por Microscopia Confocal Combinada e Super-Resolução

Este protocolo fornece uma abordagem padronizada para imagens da morfologia mitocondrial em múltiplos tecidos de C. elegans durante o envelhecimento.

Imagem e Quantificação da Morfologia Mitocondrial em C. elegans Durante o Envelhecimento

O protocolo a seguir descreve o processo de dissecção pancreática para imagens fatia virtual, e posterior quantificação de todos os GFP-tagged células beta no pâncreas inteiro.

Quantificação em situ da Missa das células beta pancreáticas em camundongos

Aprenda a quantificar a morfologia mitocondrial in situ em músculo esquelético e células de Schwann terminais de camundongos usando microscopia confocal tridimensional. Este método permite analisar redes mitocondriais em tecidos intactos sem necessidade de repórteres transgênicos.

Métodos para Quantificação In Situ da Morfologia Mitocondrial em Células Musculares e Terminais de Schwann de Camundongos

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