Chapter 16. Two-photon microscopy and multidimensional analysis of cell dynamics

Area of Science:

• * Biomedical imaging
• * Cell biology
• * Immunology

Background:

• * Two-photon (2P) microscopy is a high-resolution imaging technique.
• * Initially used in neurobiology and developmental cell biology, it is now widely adopted in immunology.
• * 2P microscopy provides exceptional views of single-cell spatiotemporal dynamics within intact tissues and organs.

Purpose of the Study:

• * To discuss the technical aspects of 2P microscope design.
• * To explain various tissue imaging preparations for 2P microscopy.
• * To guide readers through the analysis and presentation of multidimensional data sets.

Main Methods:

• * Review of 2P microscope design principles.
• * Detailed explanation of tissue preparation techniques for imaging.
• * Guidance on multidimensional data analysis and result presentation.

Main Results:

• * 2P microscopy enables unparalleled visualization of cellular dynamics in vivo.
• * Advancements in technology and reagents enhance its utility.
• * Comprehensive understanding of technical, preparative, and analytical aspects is crucial for effective application.

Conclusions:

• * Two-photon microscopy is an increasingly valuable tool for cell function assessment and molecular mechanism studies.
• * The technology's development, coupled with new reagents, expands its research potential.
• * This review serves as a comprehensive resource for researchers utilizing 2P microscopy.