Cilia : Methods and Protocols / edited by Vito Mennella

Combination of CRISPR-Cas9-RNP and Single-Cell RNAseq to Identify Cell State-Specific FOXJ1 Functions in the Human Airway Epithelium -- SARS-CoV-2 Infection of Human Primary Nasal Multiciliated Epithelial Cells Grown on Air-Liquid Interface Cultures -- A Chemically Inducible Organelle Rerouting Assay to Probe Primary Cilium Assembly, Maintenance, and Disassembly in Cultured Cells -- Expansion Microscopy of Ciliary Proteins -- Immunolabel-First-Expand-Later Expansion Microscopy Approach using Stable STED Dyes -- Structural Analysis of Sperm Centrioles Using N-STORM -- A Novel Sandwich Method for Serial Block Face SEM Imaging of Airway Multiciliated Epithelium -- Airway Cells 3D Reconstruction via Manual and Machine-Learning Aided Segmentation of Volume EM Datasets -- Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging -- Live-Imaging Centriole Amplification in Mouse Brain Multiciliated Cells -- Proximity Mapping of Ciliary Proteins by BioID -- Affinity Purification of Intraflagellar Transport (IFT) Proteins in Mice using Endogenous Streptavidin/FLAG Tags -- Primary Human Nasal Epithelial Cell Culture -- BMI1 Transduction of Human Airway Epithelial Cells for Expansion of Proliferation and Differentiation -- High Speed Video Microscopy of Ependymal Cilia in Brain Organotypic and Cell Culture Models -- Measuring Biophysical Properties of Cilia Motility from Mammalian Tissues via Quantitative Video Analysis Methods -- Mucociliary Transport Device Construction and Application to Study Mucociliary Clearance..

This volume covers the latest advancements in the study of ciliary complexity. Protocols cover genomic, proteomic, imaging, and functional analysis of different ciliated tissues and their wide applicability in cilia biology. Chapters in this book primarily focus on methods to study multiciliated cells, and discuss topics such as SARS-CoV-2 infections of human primary nasal multiciliated epithelial cells; expansion microscopy of ciliary proteins; live-imaging centriole amplification in mouse brain multiciliated cells; biophysical properties of cilia motility; and mucociliary transport device construction. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Cilia: Methods and Protocols is a valuable resource for researchers who are interested in learning more about this developing field. ..

Medienart:

E-Book

Erscheinungsjahr:

2024.

Erschienen:

New York, NY: Springer US ; 2024.

New York, NY: Imprint: Humana ; 2024.

Ausgabe:

1st ed. 2024.

Reihe:

Methods in Molecular Biology - 2725

Sprache:

Englisch

Beteiligte Personen:

Mennella, Vito [HerausgeberIn]

Links:

doi.org [lizenzpflichtig]

ISBN:

978-1-0716-3507-0

Themen:

Cell organelles.
Cytology.

Umfang:

1 Online-Ressource(XII, 278 p. 56 illus., 50 illus. in color.)

doi:

10.1007/978-1-0716-3507-0

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

1867446006