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Building a cell from its component parts /
This new volume of Methods in Cell Biology looks at building a cell from its component parts. Chapters cover such topics as engineering motor scaffolds, artificial cytoskeletons, interconnected droplet networks and artificial cells; building cytoskeletal systems and artificial actin cortex on pillar...
| Clasificación: | Libro Electrónico |
|---|---|
| Otros Autores: | , |
| Formato: | Electrónico eBook |
| Idioma: | Inglés |
| Publicado: |
Amsterdam :
Academic Press,
2015.
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| Edición: | First edition. |
| Colección: | Methods in cell biology ;
v. 128. |
| Temas: | |
| Acceso en línea: | Texto completo Texto completo |
Tabla de Contenidos:
- Front Cover; Methods in Cell Biology; Series Editors; Methods in Cell Biology; Copyright; Contents; Contributors; Preface; 1. In vitro systems for the study of microtubule-based cell polarity in fission yeast; Introduction; 1. Rationale; 2. Materials; 2.1 Materials for Method 1: Elongated Glass Wells with TiO2 Overhang; 2.1.1 Microfabrication; 2.1.2 Surface functionalization; 2.1.3 Assay; 2.2 Materials for Method 2: Elongated Water in Oil Emulsion Droplets; 2.2.1 Microfluidic chip; 3. Methods; 3.1 Method 1: Elongated Glass Wells with TiO2 Overhang; 3.1.1 Design.
- 3.1.2 Deposition of chromium and titanium oxide by electron-beam evaporation3.1.3 Postannealing of titanium oxide; 3.1.4 UV-lithography with S1813; 3.1.5 Reactive ion etching; 3.1.6 Wet etching with KOH; 3.1.7 Surface functionalization; 3.1.8 Assays; 3.2 Method 2: Elongated Water in Oil Emulsion Droplets; 3.2.1 Microfluidic design; 3.2.2 Masks for microfluidic chip fabrication; 3.2.3 SU-8 negative mould; 3.2.4 PDMS chip (Figure 5); 3.2.5 Microfluidic setup and droplet formation; 3.2.6 Lipids composition; 3.2.7 Assays; Discussion and Perspectives; Acknowledgments; References.
- 2. Microtubules, MAPs, and motor patternsIntroduction; 1. Methods; 2. Simple Filament-gliding Assay with Kinesin-1 and Microtubules; 2.1 Flow Chambers; 2.1.1 Materials; 2.1.2 Chamber construction; 2.1.3 Notes; 2.2 Gliding Assay Reagents and Buffers; 2.3 Experimental Details; 2.4 Analysis and Notes; 2.4.1 Kymographs; 2.4.2 Filament end tracking; 2.4.3 Notes; 3. Gliding Assay with MAP65 Cross-linkers to Visualize Dynamic Cross-linking; 3.1 MAP65 Purification; 3.1.1 Preparation reagents and buffers; 3.1.2 Protein purification; 3.1.2.1 Day 1; 3.1.2.2 Day 2; 3.1.2.3 Day 3.
- 3.2 Gliding Assay Buffers3.3 Experimental Details; 3.4 Analysis and Notes; 4. Cell-like Patterns from Gliding Prebundled Microtubule Filaments; 4.1 Reagents and Buffers; 4.2 Prebundling Microtubules; 4.3 Experimental Details; Discussion and Summary; Acknowledgments; References; 3. Self-organization of motors and microtubules in lipid-monolayered droplets; Introduction; 1. Materials; 1.1 Equipment; 1.1.1 Glass surface treatment and Polydimethylsiloxane (PDMS) chamber preparation; 1.1.2 Lipid-oil mixture preparation; 1.1.3 Buffer droplet in oil production; 1.1.4 Confocal fluorescence microscopy.
- 1.2 Reagents1.2.1 For glass surface treatment and PDMS chamber preparation; 1.2.2 For lipid-oil mixture preparation and buffer droplet in oil production; 1.3 Buffers and Solutions; 1.4 Proteins Used in the Assays; 2. Methods; 2.1 Glass Surface Treatment and PDMS Chamber Preparation; 2.1.1 Cleaning and hydrophobic surface treatment; 2.1.2 PDMS chambers; 2.2 Lipid-Oil Mixture Preparation; 2.3 Buffer Droplet in Oil Production; 2.3.1 Droplet solution preparation; 2.3.2 Resuspension of droplets in oil; 2.4 Confocal Fluorescence Microscopy; Results and Discussion; Acknowledgments; References.