Name: 마우스의 근육 및 말단 슈반 세포에서 미토콘드리아 형태의 현장 정량화 방법 (동영상) | JoVE Journal
Uploaded: 2026-04-10T13:00:00.000+00:00
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Description: 배우세요 마우스 골격근과 말단 슈반 세포에서 미토콘드리아 형태를 현장 정량화하는 방법을. 막전위 민감성 염료와 공초점 현미경을 활용하여 신경근육 조직 내 미토콘드리아 네트워크를 3차원으로 분석할 수 있습니다.

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<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>
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<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>
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<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>

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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>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 구동 이미지 분석 소프트웨어</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>미토콘드리아 염색 재구성 시약</td></tr><tr><td>Hoechst 33342</td><td>Thermo Fisher Scientific</td><td>H1399</td><td>세포 투과성 핵산 염색</td></tr><tr><td>ImageJ/Fiji&nbsp;</td><td>National Institutes of Health, Bethesda, MD, USA</td><td>SCR_002285</td><td>2D 분석 소프트웨어</td></tr><tr><td>Invitrogen ProLong Gold antifade reagent with DAPI&nbsp;</td><td>Fisher Scientific, Hampton, NJ, USA</td><td>P36941</td><td>실장 매체</td></tr><tr><td>KimWipe</td><td>Kimberly-Clark, Irving, TX, USA</td><td>06-666A</td><td>실험실 물티슈</td></tr><tr><td>Leica LAS_X software&nbsp;</td><td>Leica Biosystems, Wetzlar, Germany</td><td>SCR_013673</td><td>공초점 소프트웨어</td></tr><tr><td>MitoView Fix 640</td><td>Biotium</td><td>70075-50ug</td><td>Far-red 형광 미토콘드리아 염색</td></tr><tr><td>NIS-Elements Advanced Research software&nbsp;</td><td>Nikon, Melville, NY, USA</td><td>SCR_014329</td><td>단면적 분석 소프트웨어</td></tr><tr><td>Permount Mounting Medium&nbsp;</td><td>Fisher Scientific, Waltham, MA 02454, USA</td><td>#SP15-100</td><td>일반 실장 매체</td></tr><tr><td>Prism 9 software&nbsp;</td><td>GraphPad Software, La Jolla, CA, USA</td><td>RRID: SCR_002798</td><td>통계 분석 소프트웨어</td></tr><tr><td>Rhodamine (TRITC)</td><td>Fisher Scientific; Hampton, NJ, USA</td><td>AP132RMI</td><td>2차 항체</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>공초점 현미경</td></tr><tr><td>SZ61</td><td>Olympus, Breinigsville, PA, USA</td><td>sz61</td><td>입체 현미경</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>최적 절단 온도 화합물</td></tr><tr><td>Visikol I</td><td>Sigma-Aldrich, St. Louis, MO, USA</td><td>H1-30</td><td>조직 클리어링 용액 1</td></tr><tr><td>Visikol II</td><td>Sigma-Aldrich, St. Louis, MO, USA</td><td>H2-30</td><td>조직 클리어링 용액 2</td></tr><tr><td>&alpha;-bungarotoxin 552</td><td>Biotrend, Koln, Germany</td><td>00014</td><td>아세틸콜린 수용체 염료</td></tr></tbody></table>
```

마우스의 근육 및 말단 슈반 세포에서 미토콘드리아 형태의 현장 정량화 방법

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

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

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Research

연구

JoVE Journal

Biology

생물학

이러한 신경 근육 접합 (NMJs)에서 터미널 Schwann 세포 (SCS)과 같은 밀도가 높은 포장 조직에 개별 셀을 표시하는 것은 도전입니다. "선형 사진 표백"표백 순서는 시간 경과 영상과 결합 할 수있는 triangularis sterni의 근육 explant, 편리하게 신경 근육 준비에 예를 들어, 단일 터미널 SCS를 delineating 수...

연속 신경 근육 분기점에서 싱글 Schwann 세포를 윤곽을 그리다하는 사진 표백

골격 근육은 운동을 위해 필수적이며, 몸의 주요 단백질 저장소입니다. C. elegans의 내 근육 건강 측정이 설명되어 있습니다. 근육의 구조와 기능에 대한 전향 적 변화는 지역화 된 GFP 및 양이온 성 염료를 사용하여 평가됩니다.

방법은 근육의 세포 내 구획을 평가하기 위해 C. 엘레

이 프로토콜을 시각화 하 고 신경 특정 미토 콘 드리 아를 계량 3 차원 (3D) 영상 및 분석 기법을 사용 합니다. 기술 한 형광 신호를 다른 형광 신호에서 데이터의 하위 집합을 분리 하는 데 사용은 다른 상황에 적용 됩니다.

3 차원 영상 및 인간 Intraepidermal 신경 섬유 내 미토 콘 드리 아의 분석

포유류 뇌 조직에서 미토콘드리아 시각화 및 분석은 어려운 작업입니다. 여기에서는 연속 블록면 주사 전자 현미경(SBFSEM)의 3차원(3D) 재구성 분석을 사용하여 이 중요한 에너지 생성 소기관의 형태학적 및 체적 분석에 대한 통찰력을 얻는 방법을 설명합니다.

직렬 블록 얼​​굴 스캐닝 전자 현미경을 사용하여 뇌 미토콘드리아 분석

우리는 아스코르바테 과산화수역 2 및 고추냉이 과산화증 염색을 포함하는 상관경 및 부피 전자 현미경을 사용하여 유전자 변형 후 전체 세포에서 미토콘드리아 및 호반 망상염식의 분포를 연구하는 프로토콜을 제시하고, 동일한 섹션에 있는 표적 유전자의 유무에 관계없이 세포의 직렬 단면도, 및 전자 현미경 검사법을 통해 직렬 화상 진찰.

미토콘드리아 및 코피포지티컬 라이트 및 부피 전자 현미경에 의한 산후성 망상 이미징

이 연구는 자유롭게 사용할 수있는 이미지 처리 도구를 사용하여 C. elegans에서 시냅스 크기와 현지화, 근육 형태 및 미토콘드리아 모양의 정량적 측정을 설명합니다. 이 접근은 C. elegans에 있는 미래 연구 결과가 유전 돌연변이 결과로 조직과 세포기관 구조 변경의 넓이를 정량적으로 비교하는 것을 허용합니다.

예쁜꼬마선충의 세포구조와 세포세포형태를 연구하기 위한 정량적 접근법

신경 근육 접합 구성 요소를 정확하게 검출하는 능력은 병리학적 또는 발달 과정으로 인해 아키텍처의 수정을 평가하는 데 중요합니다. 여기서 우리는 정량 적 측정을 수행하는 데 사용할 수있는 전산 신경 근육 접합의 고품질 이미지를 얻기 위해 간단한 방법에 대한 완전한 설명을 제시합니다.

면역형광및 전체산 신경근육 접합의 형태 분석에 대한 단일 골격 근육 섬유의 해부

이 프로토콜은 SMA 및 ColQ 관련 CMS의 마우스 모델에서 병리학 적 변화를 정량화하는 데 사용되는 컨 포칼 및 STED 현미경을 결합하여 신경근 접합부의 형태 측정 분석 방법을 설명합니다.

결합된 공초점 및 초분해능 현미경에 의한 마우스의 신경근 접합부 특성 분석

이 프로토콜은 노화 중 예쁜꼬마선충 의 여러 조직에서 미토콘드리아 형태를 이미징하기 위한 표준화된 접근 방식을 제공합니다.

Imaging and Quantifying Mitochondrial Morphology in C. elegans during aging (노화 중 예쁜꼬마선충의 미토콘드리아 형태학의 이미징 및 정량화)

다음 프로토콜은 가상 슬라이스 이미지에 대한 췌장 해부 과정을 설명하고, 전체 췌장에있는 모든 GFP - 태그 베타 세포의 후속 부량.

마우스의 췌장 베타 세포 질량의 원위치 부량에서

배우세요 마우스 골격근과 말단 슈반 세포에서 미토콘드리아 형태를 현장 정량화하는 방법을. 막전위 민감성 염료와 공초점 현미경을 활용하여 신경근육 조직 내 미토콘드리아 네트워크를 3차원으로 분석할 수 있습니다.

마우스의 근육 및 말단 슈반 세포에서 미토콘드리아 형태의 현장 정량화 방법

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