An automated high-throughput counting method for screening circulating tumor cells in peripheral blood

Area of Science:

• Biomedical Engineering
• Cancer Research
• Microfluidics

Background:

• Enumeration of circulating tumor cells (CTCs) is crucial for cancer diagnosis and prognosis.
• Existing methods for CTC detection face limitations in throughput and speed.

Purpose of the Study:

• To develop and validate an automated, high-throughput method for enumerating CTCs.
• To compare the performance of the new method against the established CellSearch assay.

Main Methods:

• Utilized microfluidics for sample handling and line-confocal microscopy for cell interrogation.
• Employed direct blood labeling with multiple fluorescently conjugated antibodies.
• Automated CTC counting based on fluorescence signals and specific labeling patterns.

Main Results:

• Achieved a high recovery rate of 94% for cultured breast cancer cells.
• Demonstrated a low background level of 1.2 counts/mL from healthy donors.
• Successfully enumerated CTCs in stage IV breast cancer patients, with counts ranging from 15 to 3375 per 7.5 mL.
• Identified CTCs in patients positive for Her2 or CD44(+)/CD24(-).

Conclusions:

• The developed automated method provides high-throughput and sensitive CTC enumeration.
• This technique shows potential for improving clinical diagnosis and prognosis in cancer patients.
• The method's speed (analyzing 1 mL in <30 min) and accuracy offer significant clinical utility.