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Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers
Published on: April 9, 2014
Complete Visualization of T Follicular Helper Cells in Germinal Centers by Light Sheet Fluorescence Microscopy
Liat Stoler-Barak1, Ziv Shulman2
1Department of Immunology, Weizmann Institute of Science, Rehovot, Israel.
Insights
A new whole-organ imaging method visualizes antigen-specific T and B cells in germinal centers (GCs). This approach overcomes limitations of current techniques, enabling detailed 3D mapping of immune cell distribution for better understanding of long-lasting immunity.
Area of Science:
- Immunology
- Cell Biology
- Microscopy
Background:
- Long-lasting immunity relies on antibody-forming cells generated within germinal centers (GCs).
- Conventional imaging methods like immunohistology and intravital microscopy have limitations in capturing the full scope of GC niches and cellular dynamics.
- Two-photon laser scanning microscopy (TPLSM) offers large-area tissue scanning but is limited by penetration depth, hindering comprehensive visualization of all GC niches.
Purpose of the Study:
- To develop and present a novel large-scale imaging method for visualizing immune cells within entire organs.
- To overcome the limitations of existing techniques in capturing the spatial distribution and quantity of immune cells in all GC niches.
- To enable a more thorough understanding of immune cell localization and interactions crucial for adaptive immunity.
Main Methods:
- Development of a large-scale imaging approach to visualize antigen-specific T and B cells.
- Application of the method to study immune cell distribution across multiple microanatomical locations and niches within lymphoid organs.
- Creation of detailed 3D maps to quantify immune cell populations and their spatial organization.
Main Results:
- The new method successfully visualizes antigen-specific T and B cells in diverse microanatomical locations within lymphoid organs.
- It allows for the capture of immune cell niches that are often missed by conventional imaging techniques.
- Detailed 3D spatial maps of immune cell locations and quantities can be generated.
Conclusions:
- This large-scale imaging approach significantly enhances the ability to study immune cell distribution in lymphoid organs.
- It provides a more comprehensive view of germinal center niches and cellular organization compared to existing methods.
- The technique holds promise for advancing our understanding of the cellular basis of long-lasting immunity.
Abstract:
Long-lasting immunity depends on generation of antibody forming cells in germinal centers (GCs). Conventional methods such as immunohistology and intravital live imaging have been used extensively to investigate the location of cellular assemblies within tissues as well as their dynamic motility and cellular interactions. Two photon laser scanning microscopy (TPLSM) intravital imaging allows scanning of large areas within tissues and reveals multiple immune cell niches. Nonetheless, this type of imaging is limited by the depth of penetration and cannot capture effectively all of the GC niches within lymphoid organs. Here we describe a method to visualize antigen-specific T and B cells in multiple microanatomical locations and niches at the level of a whole organ. This large-scale imaging approach can greatly increase our understanding of the spatial distribution of immune cells and help obtain detailed 3D maps of their locations and quantities.
