Improved diffuse fluorescence flow cytometer prototype for high sensitivity detection of rare circulating cells in...

Area of Science:

• Biomedical Engineering
• Preclinical Research
• Cellular Biology

Background:

• Detecting rare circulating cells in mice is crucial for preclinical biomedical research.
• Existing in vivo flow cytometry methods have limitations in blood sampling volume and sensitivity.
• Diffuse fluorescence flow cytometry (DFFC) was previously developed to address these limitations.

Purpose of the Study:

• To introduce a novel DFFC prototype with significantly improved sensitivity.
• To detail the technical innovations enabling enhanced performance.
• To validate the system's capability for accurate in vivo cell enumeration.

Main Methods:

• Development of a new DFFC prototype incorporating technical innovations.
• Implementation of a motion artifact removal technique for hindleg interrogation.
• Utilized improved collection optics and signal preamplification.
• Validation using optical flow phantoms and fluorescently labeled mesenchymal stem cells in nude mice.

Main Results:

• The new DFFC prototype demonstrates an order-of-magnitude improvement in sensitivity over previous versions.
• Achieved an overall cell counting sensitivity of approximately 1 cell/mL or better in vivo.
• Successfully detected and enumerated fluorescently labeled mesenchymal stem cells at low concentrations.

Conclusions:

• The enhanced DFFC system offers a powerful tool for sensitive detection and enumeration of rare circulating cells.
• This advancement significantly improves capabilities in preclinical biomedical research.
• The system's high sensitivity and accuracy facilitate better understanding of cellular dynamics in vivo.