Name: Farelerin Kas ve Terminal Schwann Hücrelerinde Mitokondriyal Morfolojinin Yerinde Niceletilmesi Yöntemleri (Video) | JoVE Journal
Uploaded: 2026-04-10T13:00:00.000+00:00
Duration: 8 min 13 s
Description: Öğrenin farelerin iskelet kası ve terminal Schwann hücrelerinde mitokondriyal morfolojiyi yerinde niceletme yöntemini. Membran potansiyeline duyarlı boya ve konfokal mikroskopi kullanarak üç boyutlu ağ yapısını analiz edin.

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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>Analiz yazılımı ile kas fonksiyon sistemi</td></tr><tr><td>AIVIA&nbsp;</td><td>Leica Microsystems, Bellevue, WA, USA</td><td>15</td><td>AI destekli görüntü analiz yazılımı</td></tr><tr><td>anti-laminin&nbsp;</td><td>MilliporeSigma</td><td>L9393-.2mL</td><td>Miyofibril sınır etiketi</td></tr><tr><td>B6; D2-Tg(S100-EGFP)1Wjt/J Fare Suşu</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 Fare Suşu</td><td>Jackson Labs, Bar Harbor, ME</td><td>JAX:13141</td><td>D2.mdx</td></tr><tr><td>DBA/2J Fare Suşu</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>Mitokondri boyama yeniden yapılandırma reaktifi</td></tr><tr><td>Hoechst 33342</td><td>Thermo Fisher Scientific</td><td>H1399</td><td>Hücre geçirgen nükleik asit boyası</td></tr><tr><td>ImageJ/Fiji&nbsp;</td><td>National Institutes of Health, Bethesda, MD, USA</td><td>SCR_002285</td><td>2D analiz yazılımı</td></tr><tr><td>Invitrogen ProLong Gold antifade reaktifi ile DAPI&nbsp;</td><td>Fisher Scientific, Hampton, NJ, USA</td><td>P36941</td><td>Montaj ortamı</td></tr><tr><td>KimWipe</td><td>Kimberly-Clark, Irving, TX, USA</td><td>06-666A</td><td>Laboratuvar mendil</td></tr><tr><td>Leica LAS_X yazılımı&nbsp;</td><td>Leica Biosystems, Wetzlar, Germany</td><td>SCR_013673</td><td>Konfokal yazılım</td></tr><tr><td>MitoView Fix 640</td><td>Biotium</td><td>70075-50ug</td><td>Uzak kırmızı floresan mitokondriyal boya</td></tr><tr><td>NIS-Elements Advanced Research yazılımı&nbsp;</td><td>Nikon, Melville, NY, USA</td><td>SCR_014329</td><td>Kesitlerin kesit alanı analiz yazılımı</td></tr><tr><td>Permount Montaj Ortamı&nbsp;</td><td>Fisher Scientific, Waltham, MA 02454, USA</td><td>#SP15-100</td><td>Genel montaj ortamı</td></tr><tr><td>Prism 9 yazılımı&nbsp;</td><td>GraphPad Software, La Jolla, CA, USA</td><td>RRID: SCR_002798</td><td>İstatistiksel analiz yazılımı</td></tr><tr><td>Rhodamine (TRITC)</td><td>Fisher Scientific; Hampton, NJ, USA</td><td>AP132RMI</td><td>İkincil antikor</td></tr><tr><td>Stellaris 5 Beyaz Işık Konfokal Mikroskobu ve ilgili dekonvolüsyon yazılımı</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>Konfokal mikroskop</td></tr><tr><td>SZ61</td><td>Olympus, Breinigsville, PA, USA</td><td>sz61</td><td>Stereo mikroskop</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 kesme sıcaklığı bileşiği</td></tr><tr><td>Visikol I</td><td>Sigma-Aldrich, St. Louis, MO, USA</td><td>H1-30</td><td>Dokunun berraklaştırma çözeltisi 1</td></tr><tr><td>Visikol II</td><td>Sigma-Aldrich, St. Louis, MO, USA</td><td>H2-30</td><td>Dokunun berraklaştırma çözeltisi 2</td></tr><tr><td>&alpha;-bungarotoxin 552</td><td>Biotrend, Koln, Germany</td><td>00014</td><td>Asetilkolin reseptör boyası</td></tr></tbody></table>

farelerin kas ve terminal schwann hücrelerinde mitokondriyal morfolojinin yerinde niceletilmesi yöntemleri

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

Farelerin Kas ve Terminal Schwann Hücrelerinde Mitokondriyal Morfolojinin Yerinde Niceletilmesi Yöntemleri

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

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Gibi nöromüsküler kavşaklar (NMJs) terminal Schwann hücreleri (SC'ler) gibi sık donatılı dokularda, tek tek hücrelerin Görselleştirme zordur. "Ardışık fotoğraf beyazlatma" bleaching sıralı time-lapse görüntüleme ile kombine edilebilir triangularis sternumun kas eksplant, elverişli bir sinir-kas preparatı,, örneğin tek bir terminal SC'ler, tasvir verir...

Sıralı Nöromusküler Kavşağı&#39;nda Tek Schwann Hücreleri belirginleştiren Photo-beyazlatma

İskelet kası hareket etmek için gerekli olan ve bedenimizin ana protein deposudur. C. elegans içinde kas sağlık ölçüleri tanımlanmıştır. Kas yapısı ve fonksiyonu için Aday değişiklikler lokalize GFP ve katyonik boyalar kullanılarak değerlendirilir.

Yöntemler içinde Muscle hücrealtı Kompartmanları DeğerlendirecekÜSKÜP ° C. elegans

Bu iletişim kuralı görselleştirmek ve sinir özgü mitokondri ölçmek için üç boyutlu (3D) görüntüleme ve çözümleme teknikleri kullanır. Teknikleri bir floresan sinyal başka bir floresan sinyalinden gelen verilerin bir alt kümesini yalıtmak için kullanıldığı diğer durumlar için geçerlidir.

Üç boyutlu görüntüleme ve mitokondri içinde insan Intraepidermal sinir lifleri analizi

Memeli beyin dokusundan mitokondriyal görselleştirme ve analiz zorlu bir iştir. Burada, bu kritik enerji üreten organelin morfolojik ve hacimsel analizi hakkında bilgi edinmek için seri blok yüz taramalı elektron mikroskobundan (SBFSEM) elde edilen üç boyutlu (3D) rekonstrüksiyon analizinin nasıl kullanılabileceğini açıklıyoruz.

Seri Blok-Yüz Taramalı Elektron Mikroskop Kullanarak Beyin Mitokondri Analizi

Genetik modifikasyondan sonra tüm hücrelerde mitokondri ve endoplazmik retiplinin dağılımını incelemek için bir protokol sunuyoruz, askorbat peroksidaz 2 ve horserpu peroksidaz boyama da dahil olmak üzere bağıntılı ışık ve hacim Elektron Mikroskopisi kullanılarak aynı bölümde hedef geni olan ve olmayan hücrelerin seri bölümlemesi ve elektron mikroskobu ile seri görüntüleme.

Göreli ışık ve hacim elektron mikroskobu ile mitokondri ve endoplazmik reticulum görüntüleme

Bu çalışmada, sinaptik boyut ve lokalizasyon, kas Morfoloji ve marochondrial şekil nicel ölçümler serbestçe kullanılabilir görüntü işleme araçları kullanarak C. elegans özetliyor. Bu yaklaşım, C. elegans 'da gelecekteki çalışmaların, genetik mutasyonların bir sonucu olarak doku ve organel yapısal değişikliklerin kapsamını nicel olarak karşılaştırmasına olanak sağlar.

Caenorhabditis elegans hücresel yapıları ve organelle morfoloji eğitim için nicel yaklaşımlar

Nöromüsküler kavşak bileşenlerini doğru bir şekilde tespit edebilme yeteneği, patolojik veya gelişimsel süreçler nedeniyle mimarisindeki değişikliklerin değerlendirilmesinde çok önemlidir. Burada, nicel ölçümler yapmak için kullanılabilecek tam montajlı nöromüsküler kavşakların yüksek kaliteli görüntülerini elde etmek için basit bir yöntemin tam bir açıklamasını sunuyoruz.

Tüm Mount Nöromüsküler Kavşakların İmmünofluoresan ve Morfometrik Analizleri için Tek İskelet Kas Liflerinin Diseksiyonu

Bu protokol, SMA ve ColQ ile ilişkili CMS'nin fare modellerindeki patolojik değişiklikleri ölçmek için kullanılan kombine konfokal ve STED mikroskopisi ile nöromüsküler kavşakların morfometrik analizi için bir yöntemi açıklamaktadır.

Farelerde Nöromüsküler Kavşakların Kombine Konfokal ve Süper Redüksiyonlu Mikroskopi ile Karakterizasyonu

Bu protokol, yaşlanma sırasında C. elegans'ın çoklu dokularında mitokondriyal morfolojiyi görüntülemek için standart bir yaklaşım sağlar.

Yaşlanma Sırasında C. elegans'ta Mitokondriyal Morfolojinin Görüntülenmesi ve Ölçülmesi

Sanal kesit görüntüleme için aşağıdaki protokol pankreas diseksiyonu sürecini özetliyor ve sonraki tüm pankreas tüm GFP etiketli beta-hücrelerinin kantifikasyon.

Farelerde, pankreatik beta-hücre Kütle yerinde Niceleme

Öğrenin farelerin iskelet kası ve terminal Schwann hücrelerinde mitokondriyal morfolojiyi yerinde niceletme yöntemini. Membran potansiyeline duyarlı boya ve konfokal mikroskopi kullanarak üç boyutlu ağ yapısını analiz edin.

Farelerin Kas ve Terminal Schwann Hücrelerinde Mitokondriyal Morfolojinin Yerinde Niceletilmesi Yöntemleri

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