Mean and variance of ratio estimators used in fluorescence ratio imaging

Cytometry. 2000 Apr 1;39(4):300-5. doi: 10.1002/(sici)1097-0320(20000401)39:4<300::aid-cyto8>3.0.co;2-o.

Abstract

Background: The ratio of two measured fluorescence signals (called x and y) is used in different applications in fluorescence microscopy. Multiple instances of both signals can be combined in different ways to construct different ratio estimators.

Methods: The mean and variance of three estimators for the ratio between two random variables, x and y, are discussed. Given n samples of x and y, we can intuitively construct two different estimators: the mean of the ratio of each x and y and the ratio between the mean of x and the mean of y. The former is biased and the latter is only asymptotically unbiased. Using the statistical characteristics of this estimator, a third, unbiased estimator can be constructed.

Results: We tested the three estimators on simulated data, real-world fluorescence test images, and comparative genome hybridization (CGH) data. The results on the simulated and real-world test images confirm the presented theory. The CGH experiments show that our new estimator performs better than the existing estimators.

Conclusions: We have derived an unbiased ratio estimator that outperforms intuitive ratio estimators.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Computer Simulation
  • Fluorescent Dyes / analysis*
  • Gene Dosage
  • Image Processing, Computer-Assisted* / methods
  • Image Processing, Computer-Assisted* / statistics & numerical data
  • Microscopy, Fluorescence / methods
  • Microscopy, Fluorescence / statistics & numerical data
  • Normal Distribution
  • Nucleic Acid Hybridization*
  • Poisson Distribution

Substances

  • Fluorescent Dyes