Three-Dimensional Patterning of the ECM Microenvironment Using Magnetic Nanoparticle Self Assembly [0.03%]
基于磁性纳米粒子自组装的三维微环境构象技术
Jiyun Kim,Kandice Tanner
Jiyun Kim
This protocol describes a way to introduce topography to three-dimensional (3D) biomaterials. The self-assembling behavior of magnetic particles can be exploited to form nanoscale to microscale fibers, such that one can dissect the contribu...
Analysis of the Stromal Cellular Components of the Solid Tumor Microenvironment Using Flow Cytometry [0.03%]
流式细胞术分析实性肿瘤微环境的间质细胞成分
Michael Timaner,Ofrat Beyar-Katz,Yuval Shaked
Michael Timaner
The tumor microenvironment consists of a variety of cell types. The contribution of each cell type to the tumor is an emerging subject in the field of cancer research. Here, we describe protocols for dissociating tumor tissues and Matrigel ...
Preparation of High-Density Fibrillar Collagen Matrices That Mimic Desmoplastic Tumor Stroma [0.03%]
仿生肿瘤周细胞的高密度纤维素基质的制备
Vira V Artym
Vira V Artym
The stroma of invasive tumors becomes enriched in dense fibrillar collagen as a result of the desmoplastic reaction. This desmoplastic collagen exerts profound effects on tumor and normal cells. In view of these findings, it is important to...
Immunofluorescence Staining [0.03%]
免疫荧光染色技术
Julie G Donaldson
Julie G Donaldson
This unit provides a protocol for indirect immunofluorescence, which is a method that provides information about the locations of specific molecules and the structure of the cell. Antibody molecules for a specific target molecule are expose...
John M Graham
John M Graham
This unit provides both a theoretical and a practical background to all the techniques associated with the application of differential and density gradient centrifugation for the analysis of subcellular membranes. The density gradient infor...
A Novel Assay to Identify the Trafficking Proteins that Bind to Specific Vesicle Populations [0.03%]
一种识别与特定囊泡群结合的蛋白质的新测定法
Marvin Bentley,Gary Banker
Marvin Bentley
Here we describe a method capable of identifying interactions between candidate trafficking proteins and a defined vesicle population in intact cells. The assay involves the expression of an FKBP12-rapamycin binding domain (FRB)-tagged cand...
A Convenient Technique to Fix Suspension Cells on a Coverslip for Microscopy [0.03%]
一种用于显微镜检查将悬浮细胞固定在盖玻片上的便捷技术
Keiko Mihara,Tomofumi Nakayama,Hisato Saitoh
Keiko Mihara
Human myeloid HL-60 cells are usually cultured in suspension in medium containing 5% to 10% fetal bovine serum (FBS) and thus are often difficult to adhere to a coverslip. In this unit, we describe how removal of FBS from the culture medium...
Isolation of Endoplasmic Reticulum, Mitochondria, and Mitochondria-Associated Membrane and Detergent Resistant Membrane Fractions from Transfected Cells and from Human Cytomegalovirus-Infected Primary Fibroblasts [0.03%]
从转染细胞和人巨细胞病毒原代成纤维细胞中分离内质网、线粒体和线粒体相关膜及去垢剂抗性膜组分
Chad D Williamson,Daniel S Wong,Petros Bozidis et al.
Chad D Williamson et al.
Increasingly mechanistic virology studies require dependable and sensitive methods for isolating purified organelles containing functional cellular sub-domains. The mitochondrial network is, in part, closely apposed to the endoplasmic retic...
Travis J Crites,Michael Maddox,Kartika Padhan et al.
Travis J Crites et al.
Glass-supported lipid bilayers presenting freely diffusing proteins have served as a powerful tool for studying cell-cell interfaces, in particular, T cell-antigen presenting cell (APC) interactions, using optical microscopy. Here we expand...
Polarized Fluorescence Microscopy to Study Cytoskeleton Assembly and Organization in Live Cells [0.03%]
用于研究活细胞中细胞骨架组装和组织的偏振荧光显微镜技术
Molly McQuilken,Shalin B Mehta,Amitabh Verma et al.
Molly McQuilken et al.
The measurement of not only the location but also the organization of molecules in live cells is crucial to understanding diverse biological processes. Polarized light microscopy provides a nondestructive means to evaluate order within subc...