A quantitative method for the analysis of cell shape and locomotion.

Area of Science:

• Immunology
• Cell Biology
• Biomedical Engineering

Background:

• Leukocyte (white blood cell) locomotion is crucial for immune responses.
• Accurate quantitation of leukocyte movement is essential for understanding cellular functions and disease states.
• Existing methods for analyzing leukocyte locomotion can be time-consuming and labor-intensive.

Purpose of the Study:

• To develop and validate a rapid, semiautomated system for quantifying and analyzing leukocyte shape and locomotion.
• To assess the system's reproducibility and compare it with manual measurement methods.
• To determine chemokinesis rates for unstimulated and stimulated polymorphonuclear leukocytes (PMNs).

Main Methods:

• Utilized a phase microscope with a Vidicon camera to capture video images of moving leukocytes.
• Employed a Datacube video analog-digital converter and a Digital Equipment Corporation LSI 11/23 computer for image processing.
• Developed FORTRAN and MARCO assembly language programs for image segmentation, display, and calculation of leukocyte parameters (position, speed, direction, orientation) at 10-second intervals.

Main Results:

• Achieved high reproducibility with a correlation coefficient of 0.998 when comparing video system measurements to manual measurements.
• Successfully determined average chemokinesis rates of 12.8 micron/min for unstimulated PMNs and 18.1 micron/min for stimulated PMNs.
• Demonstrated the system's capability for rapid data analysis and multiparameter evaluation of leukocyte locomotion.

Conclusions:

• The developed semiautomated video system offers a fast and efficient method for analyzing leukocyte shape and locomotion.
• The system provides accurate and reproducible quantitative data, facilitating detailed studies of immune cell behavior.
• This technology is valuable for research requiring direct evaluation of leukocyte locomotion, including studies of inflammation and immune response.