Estimating weak ratiometric signals in imaging data. I. Dual-channel data.
نویسندگان
چکیده
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.
منابع مشابه
Estimating weak ratiometric signals in imaging data. II. Meta-analysis with multiple, dual-channel datasets.
Ratiometric fluorescent indicators are used for making quantitative measurements of a variety of physiological variables. Their utility is often limited by noise. This is the second in a series of papers describing statistical methods for denoising ratiometric data with the aim of obtaining improved quantitative estimates of variables of interest. Here, we outline a statistical optimization met...
متن کاملProper integration time of polarization signals of internetwork regions using Sunrise/IMaX data
Distribution of magnetic fields in the quiet-Sun internetwork areas has been affected by weak polarization (in particular Stokes Q and U) signals. To improve the signal-to-noise ratio (SNR) of the weak polarization signals, several approaches, including temporal integrations, have been proposed in the literature. In this study, we aim to investigate a proper temporal-integration time with which...
متن کاملTwo New FRET Imaging Measures: Linearly Proportional to and Highly Contrasting the Fraction of Active Molecules
We developed two new FRET imaging measures for intramolecular FRET biosensors, called linearly proportional (LP) and highly contrasting (HC) measures, which can be easily calculated by the fluorescence intensities of donor and acceptor as a ratio between their weighted sums. As an alternative to the conventional ratiometric measure, which non-linearly depends on the fraction of active molecule,...
متن کاملNew statistical methods enhance imaging of cameleon fluorescence resonance energy transfer in cultured zebrafish spinal neurons.
Cameleons are genetically encoded fluorescence resonance energy transfer (FRET)-based Ca(2+) indicators. Attempts to use cameleons to detect neural activity in vertebrate systems have been largely frustrated by the small FRET signal, in contradistinction to the higher signals seen in Drosophila and Caenorhabditis elegans. We have developed a statistical optimization method capable of detecting ...
متن کاملReversible ratiometric detection of highly reactive hydropersulfides using a FRET-based dual emission fluorescent probe† †Electronic supplementary information (ESI) available: Full experimental procedures, synthesis and characterization of compounds. See DOI: 10.1039/c6sc03856e Click here for additional data file.
Hydropersulfide (R–SSH) is an important class of reactive sulfur species (RSS) involved in a variety of physiological processes in mammals. A fluorescent probe capable of real-time detection of hydropersulfide levels in living cells would be a versatile tool to elucidate its roles in cell signalling and redox homeostasis. In this paper, we report a ratiometric fluorescent probe for hydropersulf...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of the Optical Society of America. A, Optics, image science, and vision
دوره 24 9 شماره
صفحات -
تاریخ انتشار 2007