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Updated: Jun 30, 2025
Author Spotlight: Unraveling Bacterial Responses to Antibiotics and Immune System in Tissues
Published on: March 1, 2024
High-Resolution Three-dimensional Whole-organ Tomography of Microbial Infections
Jiagui Li1, Alaa Alhayek1, Heng Wang1
1Biozentrum, University of Basel.
Abstract:
Most infections take place within three-dimensional host tissues with intricate anatomy and locally varying host physiology. The positioning of pathogen cells within this diverse environment significantly affects their stress levels, responses, fate, and contribution to the overall progression of the disease and treatment failure. However, due to the technical difficulties in locating µm-sized pathogen cells within cm-sized host organs, this area of research has been relatively unexplored. Here, we present a method for addressing this challenge. We employ serial two-photon tomography and AI-enhanced image analysis to locate individual Salmonella cells throughout the entire spleen, liver lobes, and whole lymph nodes of infected mice. Using fluorescent reporters and in vivo antibody administration, the replication rate of single Salmonella cells, their local interaction with specific immune cells, and bacterial responses to antibiotics can be determined. These methodologies open avenues for a comprehensive examination of infections, their prevention, and treatment within the three-dimensional tissue context.
Insights
Researchers developed a new method to track individual Salmonella bacteria within mouse organs. This technique helps understand infection dynamics and antibiotic effectiveness in 3D host tissues.
Area of Science:
- Microbiology
- Immunology
- Medical Imaging
Background:
- Infections occur in complex 3D host tissues, influencing pathogen behavior and treatment outcomes.
- Locating microscopic pathogens within macroscopic organs is technically challenging, limiting research.
Purpose of the Study:
- To develop and validate a method for precisely locating and analyzing individual pathogen cells within entire host organs.
- To investigate pathogen behavior, immune interactions, and antibiotic responses at the single-cell level within the 3D tissue microenvironment.
Main Methods:
- Serial two-photon tomography for high-resolution 3D imaging of entire organs.
- AI-enhanced image analysis for automated detection and tracking of individual Salmonella cells.
- Fluorescent reporters and in vivo antibody administration to assess bacterial replication, immune cell interactions, and antibiotic effects.
Main Results:
- Successfully located and analyzed individual Salmonella cells throughout the spleen, liver, and lymph nodes of infected mice.
- Quantified single-cell replication rates and characterized local interactions with host immune cells.
- Determined bacterial responses to antibiotic treatment in situ within the 3D tissue context.
Conclusions:
- The developed methodology enables unprecedented examination of pathogen dynamics within the native 3D tissue environment.
- This approach facilitates a deeper understanding of infection progression, host-pathogen interactions, and treatment efficacy.
- Opens new avenues for studying infectious diseases and developing targeted therapeutic strategies.
