System for flow sorting chromosomes on the basis of pulse shape

Area of Science:

• Cytogenetics
• Biotechnology
• Flow Cytometry

Background:

• Chromosome analysis requires sophisticated methods for classification.
• Slit-scan flow cytometry offers detailed morphological data but presents processing challenges.
• Variability in chromosome scan data necessitates robust algorithms for accurate sorting.

Purpose of the Study:

• To demonstrate the capability of sorting individual chromosomes based on morphological features using slit-scan flow cytometry.
• To develop a rapid and flexible system for pulse shape acquisition and processing for chromosome sorting.
• To enable sort decisions before droplet breakoff using complex morphological data.

Main Methods:

• Developed a sort controller for acquiring 128-byte chromosome waveforms.
• Implemented numerical computations for an area-based centromeric index sort decision within 2 ms.
• Configured the system using a NOVIX microprocessor programmed in FORTH, interfaced with a slit-scan flow cytometer data acquisition system.

Main Results:

• Successfully sorted individual chromosomes based on complex morphological features.
• Achieved sort decisions in under 2 milliseconds, enabling real-time sorting.
• Demonstrated the system's tolerance to variations in pulse shape for reliable chromosome identification.

Conclusions:

• Slit-scan flow cytometry, coupled with a rapid processing system, enables accurate chromosome sorting based on morphology.
• The developed controller offers direct control over machine state for minimal processing time.
• This technology advances chromosome classification and sorting capabilities in flow cytometry.