AQSA-Algorithm for Automatic Quantification of Spheres Derived from Cancer Cells in Microfluidic Devices

Area of Science:

• Oncology
• Bioinformatics
• Cancer Research

Background:

• Cancer stem cells (CSCs) are crucial in chemoresistance and recurrence.
• Sphere formation assays enrich CSCs but manual counting is laborious and subjective.
• Automated quantification is needed for reliable prediction chemotherapy assays.

Purpose of the Study:

• To develop and validate an automated computational program, the Automatic Quantification of Spheres Algorithm (ASQA), for detecting, counting, and measuring CSC spheres.
• To compare ASQA's performance against manual counts and existing software (SpheroidJ).
• To assess ASQA's utility in observing treatment response and its applicability in resource-limited settings.

Main Methods:

• Development of the Automatic Quantification of Spheres Algorithm (ASQA).
• Comparison of ASQA counts with manual counts (p = 0.167).
• Comparison of ASQA area measurements with SpheroidJ for single spheres (p = 0.173).
• Evaluation of ASQA performance with uniform and nonuniform backgrounds.
• Testing ASQA on LN229 cell line and primary culture CSCs.

Main Results:

• ASQA showed no statistically significant difference compared to manual counts.
• ASQA demonstrated high accuracy in area measurements, comparable to SpheroidJ.
• The algorithm efficiently detects and quantifies multiple spheres, enabling treatment response observation.
• ASQA functions effectively with uniform backgrounds; non-uniformity may introduce artifacts.
• The algorithm processes images rapidly (0.3s/image) with low computational cost.

Conclusions:

• ASQA provides an automated, accurate, and efficient method for CSC sphere quantification.
• The algorithm is adaptable for both cell lines and primary cultures.
• ASQA's accessibility and speed can benefit cancer research globally, aiding in developing better chemotherapy strategies.
• Automated CSC detection can help avoid ineffective treatments and reduce patient toxicity.