Quantitative morphometric measurements using site selective image cytometry of intact tissue

Area of Science:

• Biomedical Imaging
• Cell Biology
• Genetics

Background:

• Site-selective two-photon tissue image cytometry enables rare cell detection in 3D tissues.
• High-throughput quantification of cellular morphology using this technique remains undemonstrated.

Purpose of the Study:

• To apply site-selective two-photon tissue image cytometry for studying homologous recombination (HR) events in mouse pancreas.
• To develop and demonstrate high-throughput quantification of cellular morphological states in rare cell populations.

Main Methods:

• Utilized a site-selective two-photon tissue cytometer with a one-photon imaging subsystem to locate regions of interest containing rare recombinant cell clusters.
• Performed high-resolution 3D assays using two-photon microscopy within identified regions.
• Applied 3D morphology extraction algorithms to analyze image stacks.

Main Results:

• Successfully identified and quantified homologous recombination (HR) events in the pancreas of transgenic mice.
• Demonstrated the ability to analyze the frequency, distribution, and morphology of rare recombinant cell clusters.
• Showcased the efficiency of the combined low-magnification and high-resolution imaging approach for rare event detection.

Conclusions:

• Site-selective two-photon tissue image cytometry is effective for studying rare cellular events like HR in complex tissues.
• The developed methodology allows for detailed morphological analysis of rare cell populations, advancing high-throughput tissue cytometry.
• This approach provides insights into the spatial distribution and cellular characteristics of homologous recombination events in vivo.