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An automated cell-counting algorithm for fluorescently-stained cells in migration assays

Author(s): Al-Khazraji Baraa | Medeiros Philip | Novielli Nicole | Jackson Dwayne

Journal: Biological Procedures Online ISSN 1480-9222
Volume: 13; Issue: 1; Start page: 9; Date: 2011;
Original page

Keywords: automated cell counting | threshold | migration assays | manual cell counting

ABSTRACT
Abstract A cell-counting algorithm, developed in Matlab®, was created to efficiently count migrated fluorescently-stained cells on membranes from migration assays. At each concentration of cells used (10,000, and 100,000 cells), images were acquired at 2.5 ×, 5 ×, and 10 × objective magnifications. Automated cell counts strongly correlated to manual counts (r2 = 0.99, P < 0.0001 for a total of 47 images), with no difference in the measurements between methods under all conditions. We conclude that our automated method is accurate, more efficient, and void of variability and potential observer bias normally associated with manual counting.