Mesoscopic in vivo 3-D tracking of sparse cell populations using angular multiplexed optical projection tomography

Area of Science:

• Biomedical Imaging
• Cell Biology
• Developmental Biology

Background:

• In vivo cell tracking is crucial for understanding biological processes.
• Existing volumetric time-lapse techniques often require high light doses, potentially harming samples.
• Sparse cell distributions pose challenges for accurate 3-D localization and tracking.

Purpose of the Study:

• To develop and validate an angular multiplexed imaging technique for 3-D in vivo cell tracking.
• To improve time-lapse resolution and reduce light dose in optical projection tomography (OPT).
• To enable high-resolution mapping of cellular migration patterns in live biological samples.

Main Methods:

• Implementation of dual axis optical projection tomography (OPT) for simultaneous image acquisition at different angles.
• Utilizing angular multiplexing to achieve 3-D localization and tracking.
• Applying the technique to a live transgenic LysC:GFP zebrafish embryo model.

Main Results:

• Achieved 3-D cell tracking with time resolution matching the camera frame rate.
• Demonstrated a 200x reduction in light dose compared to conventional volumetric time-lapse OPT.
• Successfully mapped 3-D neutrophil migration patterns in a zebrafish embryo over ~25.5 minutes post-tail wound.

Conclusions:

• Angular multiplexed dual axis OPT offers superior time-lapse resolution and reduced light dose for in vivo cell tracking.
• This technique is effective for studying dynamic cellular processes like migration in live embryos.
• The method provides a valuable tool for high-resolution 3D imaging in developmental and cell biology research.