Related Experiment Video
Updated: Aug 16, 2025
A Detailed Protocol for Characterizing the Murine C1498 Cell Line and its Associated Leukemia Mouse Model
Published on: October 14, 2016
Guidelines for DC preparation and flow cytometry analysis of mouse lymphohematopoietic tissues
Lukas Amon1, Diana Dudziak1,2,3,4, Ronald A Backer5,6
1Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Erlangen, Germany.
Insights
This article provides detailed protocols for preparing single-cell suspensions from mouse tissues. These methods are crucial for analyzing dendritic cells (DCs) using flow cytometry and other applications in immunology research.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) are critical immune cells involved in initiating adaptive immune responses.
- Standardized protocols are essential for reproducible DC research.
Purpose of the Study:
- To provide detailed, state-of-the-art protocols for the preparation and analysis of mouse dendritic cells.
- To facilitate the generation of single-cell suspensions from various mouse lymphohematopoietic tissues.
Main Methods:
- Isolation of single-cell suspensions from mouse spleen, peripheral lymph nodes, and thymus.
- Phenotypic analysis of dendritic cells using flow cytometry.
- Protocols are validated by experienced scientists and peer-reviewed.
Main Results:
- Comprehensive methods for generating high-quality single-cell suspensions from mouse lymphoid organs.
- Focus on optimizing dendritic cell analysis via flow cytometry.
- Prepared cells are suitable for downstream applications like RNA sequencing and Western blotting.
Conclusions:
- This chapter offers essential, reliable protocols for researchers studying dendritic cells.
- The provided methods support both basic and clinical immunology investigations.
- Standardized DC preparation enhances the reliability of flow cytometry and other analyses.
Abstract:
This article is part of the Dendritic Cell Guidelines article series, which provides a collection of state-of-the-art protocols for the preparation, phenotype analysis by flow cytometry, generation, fluorescence microscopy, and functional characterization of mouse and human DC from lymphoid organs, and various non-lymphoid tissues. Within this chapter, detailed protocols are presented that allow for the generation of single-cell suspensions from mouse lymphohematopoietic tissues including spleen, peripheral lymph nodes, and thymus, with a focus on the subsequent analysis of DC by flow cytometry. However, prepared single-cell suspensions can be subjected to other applications including sorting and cellular enrichment procedures, RNA sequencing, Western blotting, and many more. While all protocols were written by experienced scientists who routinely use them in their work, this article was also peer-reviewed by leading experts and approved by all co-authors, making it an essential resource for basic and clinical DC immunologists.
More Related Videos
06:22Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells
Published on: January 9, 2019
09:01Flow Cytometry to Estimate Leukemia Stem Cells in Primary Acute Myeloid Leukemia and in Patient-derived-xenografts, at Diagnosis and Follow Up
Published on: March 26, 2018