In Vivo Tracking and 3D Mapping of Cell Death in Regeneration and Cancer Using Trypan Blue

Area of Science:

• Cell Biology
• Biomedical Imaging
• Pathology

Background:

• In vivo cell death tracking is crucial for understanding tissue homeostasis and disease.
• Existing methods for labeling dead cells in vivo have limitations, including poor sensitivity, tissue penetration, and temporal resolution.

Purpose of the Study:

• To develop and validate a novel fluorescent marker, Trypan Blue (TBlue), for sensitive and selective in vivo cell death detection.
• To demonstrate TBlue's utility in various biological contexts, including organ regeneration and preclinical cancer therapy assessment.

Main Methods:

• Fluorescent labeling of dead cells in vivo using Trypan Blue (TBlue).
• Application of TBlue for detecting single dead cells and mapping necrotic tissue in organ regeneration models.
• Utilizing TBlue for "pulse and chase" tracking of dying cells in regenerating mouse livers.
• Assessing TBlue's efficacy in labeling dead cancer cells and chemotherapy-induced cell death in preclinical models.

Main Results:

• TBlue effectively labeled diverse cell death types (necrosis, apoptosis) in various organs (liver, skin).
• TBlue enabled high-resolution 3D mapping of necrotic tissue during organ regeneration.
• The "pulse and chase" approach with TBlue allowed temporal tracking of dying hepatocytes.
• TBlue demonstrated utility in evaluating anticancer therapies by labeling dead cancer cells and chemotherapy-affected thymocytes.

Conclusions:

• Trypan Blue (TBlue) is a sensitive and selective marker for in vivo cell death detection.
• TBlue offers significant advantages over existing methods, including improved sensitivity, tissue penetration, and temporal resolution.
• TBlue facilitates a deeper understanding of cell death's role in normal biological processes and disease, with applications in preclinical research.