Unmixing of fluorescence spectra to resolve quantitative time-series measurements of gene expression in plate readers.

ABSTRACT:

Background

To connect gene expression with cellular physiology, we need to follow levels of proteins over time. Experiments typically use variants of Green Fluorescent Protein (GFP), and time-series measurements require specialist expertise if single cells are to be followed. Fluorescence plate readers, however, a standard in many laboratories, can in principle provide similar data, albeit at a mean, population level. Nevertheless, extracting the average fluorescence per cell is challenging because autofluorescence can be substantial.

Results

Here we propose a general method for correcting plate reader measurements of fluorescent proteins that uses spectral unmixing and determines both the fluorescence per cell and the errors on that fluorescence. Combined with strain collec

SUBMITTER: Lichten CA 

PROVIDER: S-EPMC3917901 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature