TY - JOUR
T1 - Estimating weak ratiometric signals in imaging data I. Dual-channel data
AU - Broder, Josef
AU - Majumder, Anirban
AU - Porter, Erika
AU - Srinivasamoorthy, Ganesh
AU - Keith, Charles
AU - Lauderdale, James
AU - Sornborger, Andrew
PY - 2007/9
Y1 - 2007/9
N2 - Ratiometric fluorescent indicators are becoming increasingly prevalent in many areas of biology. They are used for making quantitative measurements of intracellular free calcium both in vitro and in vivo, as well as measuring membrane potentials, pH, and other important physiological variables of interest to researchers in many subfields. Often, functional changes in the fluorescent yield of ratiometric indicators are small, and the signal-to-noise ratio (SNR) is of order unity or less. In particular, variability in the denominator of the ratio can lead to very poor ratio estimates. We present a statistical optimization method for objectively detecting and estimating ratiometric signals in dual-wavelength measurements of fluorescent, ratiometric indicators that improves on standard methods. With the use of an appropriate statistical model for ratiometric signals and by taking the pixel-pixel covariance of an imaging dataset into account, we are able to extract user-independent spatiotemporal information that retains high resolution in both space and time.
AB - Ratiometric fluorescent indicators are becoming increasingly prevalent in many areas of biology. They are used for making quantitative measurements of intracellular free calcium both in vitro and in vivo, as well as measuring membrane potentials, pH, and other important physiological variables of interest to researchers in many subfields. Often, functional changes in the fluorescent yield of ratiometric indicators are small, and the signal-to-noise ratio (SNR) is of order unity or less. In particular, variability in the denominator of the ratio can lead to very poor ratio estimates. We present a statistical optimization method for objectively detecting and estimating ratiometric signals in dual-wavelength measurements of fluorescent, ratiometric indicators that improves on standard methods. With the use of an appropriate statistical model for ratiometric signals and by taking the pixel-pixel covariance of an imaging dataset into account, we are able to extract user-independent spatiotemporal information that retains high resolution in both space and time.
UR - http://www.scopus.com/inward/record.url?scp=36448994456&partnerID=8YFLogxK
U2 - 10.1364/JOSAA.24.002921
DO - 10.1364/JOSAA.24.002921
M3 - Article
C2 - 17767264
AN - SCOPUS:36448994456
VL - 24
SP - 2921
EP - 2931
JO - Journal of the Optical Society of America A: Optics and Image Science, and Vision
JF - Journal of the Optical Society of America A: Optics and Image Science, and Vision
SN - 1084-7529
IS - 9
ER -