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Company: Sigma-Aldrich
Catalog#: H9627
Other protocol()

Muscle Histology Characterization Using H&E Staining and Muscle Fiber Type Classification Using Immunofluorescence Staining
[Abstract]  Muscle function is determined by its structure and fiber type composition. Here we describe a protocol to examine muscle histology and myofiber types using hematoxylin and eosin (H&E) and immunofluorescence staining, respectively. H&E stain nucleus in blue and cytoplasm in red, therefore allowing for morphological analyses, such as myofiber diameter, the presence of degenerated and regenerated myofibers, and adipocytes and fibrotic cells. Muscle fibers in adult skeletal muscles of rodents are classified into 4 subtypes based on the expression of myosin heavy chain proteins: Myh7 (type I fiber), Myh2 (type IIA fiber), Myh1 (type IIX fiber), Myh4 (type IIB fiber). A panel of monoclonal antibodies can be used to specifically label these muscle fiber subtypes. These protocols are commonly ... [摘要]  肌肉功能由其结构和纤维类型组成决定。 这里我们描述一个协议,分别检查使用苏木精和伊红(H&E)和免疫荧光染色的肌肉组织学和肌纤维类型。 H&E在蓝色和细胞质红色染色核,因此允许形态学分析,如肌纤维直径,退化和再生肌纤维的存在,以及脂肪细胞和纤维化细胞。 基于肌球蛋白重链蛋白Myh7(I型纤维),Myh2(IIA型纤维),Myh1(IIX型纤维),Myh4(IIB型纤维)的表达,将啮齿动物成年骨骼肌肌肉纤维分为4种亚型:。 一组单克隆抗体可用于特异性标记这些肌纤维亚型。 这些方案通常用于肌肉发育,生长和再生的研究(例如:Wang等,2015; Nie等,2016; Yue et al。,2016; Wang et al。,2017)。
【背景】骨骼肌由肌细胞,脂肪细胞,成纤维细胞和其他细胞类型组成。 多核心肌细胞,骨骼肌的主要组成,也称为肌纤维(肌纤维)。 肌肉组织学研究是肌肉功能研究的常规方法。 一般来说,肌纤维的直径和脂肪细胞的程度和纤维化面积与肌肉力量有关(Yue et al。,2016)。 此外,中心有核肌纤维的存在作为肌肉再生期间新再生肌纤维的替代指标(Wang等,2017)。 基于差异代谢性状和肌球蛋白重链(MyHC)亚型的表达,肌纤维分为四种类型(I,IIa,IIx和IIb)。 ...

Quantification of Rae-1 Staining Intensity in Glomeruli
[Abstract]  NKG2D is expressed on all NK cells and on subsets of NKT, CD8, CD4 and γδ T cells. NKG2D is activated by NKG2D ligands, a stress-induced family of MHC-I-like proteins. These ligands are upregulated on stressed/infected cells and are not widely expressed on healthy adult tissue. NKG2D ligands have been widely studied as potential targets for immunotherapeutic approaches in cancer and auto-immune diseases such as Systemic Lupus Erythematosus. Here we describe a method for quantifying the expression levels of the Rae-1 NKG2D ligand in the glomeruli of healthy and diseased individuals via a novel algorithm. The MRL mouse was used as a positive control strain as it spontaneously generates a lupus-like phenotype, one of the main effects of which is severe glomerulonephritis. MRL/MpJ mice ... [摘要]  NKG2D在所有NK细胞上和在NKT,CD8,CD4和γδT细胞的子集上表达。 NKG2D被NKG2D配体激活,NKG2D配体是应激诱导的MHC-I样蛋白家族。这些配体在应激/感染的细胞上上调,并且不在健康成人组织上广泛表达。 NKG2D配体已被广泛研究作为在癌症和自身免疫疾病如系统性红斑狼疮中的免疫治疗方法的潜在靶标。在这里我们介绍一种方法通过一个新的算法,量化健康和患病个体的肾小球Rae-1 NKG2D配体的表达水平。 MRL小鼠用作阳性对照菌株,因为其自发产生狼疮样表型,其中一个主要影响是严重的肾小球肾炎。 MRL/MpJ小鼠在约12个月龄时自发产生该表型,而具有相同遗传背景但在Fas等位基因中产生自发纯合突变的MRL/MpJlpr小鼠发展类似但更严重的症状〜3 -4个月龄(Spada等人,2015)。