An Automated Cell-counting Algorithm for Fluorescently-stained Cells in Migration Assays

Authors

Baraa K. Al-Khazraji, The University of Western Ontario
Philip J. Medeiros, The University of Western Ontario
Nicole M. Novielli, The University of Western Ontario
Dwayne N. Jackson, The University of Western Ontario

Document Type

Article

Publication Date

9-19-2011

Journal

Biological Procedures Online

Volume

13

Issue

9

URL with Digital Object Identifier

http://dx.doi.org/10.1186/1480-9222-13-9

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.