Confocal fluorescence ratiometric imaging is an optical technique used to measure a variety of important biological parameters. A small amount of chromatic aberration in the microscope system can introduce a variation in the signal ratio dependent on the fluorophore concentration gradient along the optical axis and lead to bias in the measurement. We present a theoretical model of this effect. Experimental results and simulations clearly demonstrate that this error can be significant and should not be ignored.
© 2011 Optical Society of America
Original Manuscript: July 12, 2010
Revised Manuscript: November 5, 2010
Manuscript Accepted: November 5, 2010
Published: December 27, 2010
Vol. 6, Iss. 2 Virtual Journal for Biomedical Optics
Yuxiang Lin and Arthur F. Gmitro, "Errors in confocal fluorescence ratiometric imaging microscopy due to chromatic aberration," Appl. Opt. 50, 95-102 (2011)