A reformulation of pLSA for uncertainty estimation and hypothesis testing in bio-imaging.

MOTIVATION: Probabilistic latent semantic analysis (pLSA) is commonly applied to describe mass spectra (MS) images. However, the method does not provide certain outputs necessary for the quantitative scientific interpretation of data. In particular, it lacks assessment of statistical uncertainty and the ability to perform hypothesis testing. We show how linear Poisson modelling advances pLSA, giving covariances on model parameters and supporting χ2 testing for the presence/absence of MS signal components. As an example, this is useful for the identification of pathology in MALDI biological samples. We also show potential wider applicability, beyond MS, using magnetic resonance imaging (MRI) data from colorectal xenograft models. RESULTS: Simulations and MALDI spectra of a stroke-damaged rat brain show MS signals from pathological tissue can be quantified. MRI diffusion data of control and radiotherapy-treated tumours further show high sensitivity hypothesis testing for treatment effects. Successful χ2 and degrees-of-freedom are computed, allowing null-hypothesis thresholding at high levels of confidence. AVAILABILITY AND IMPLEMENTATION: Open-source image analysis software available from TINA Vision, www.tina-vision.net. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

A new method for the high-precision assessment of tumor changes in response to treatment.

Motivation: Imaging demonstrates that preclinical and human tumors are heterogeneous, i.e. a single tumor can exhibit multiple regions that behave differently during both development and also in response to treatment. The large variations observed in control group, tumors can obscure detection of significant therapeutic effects due to the ambiguity in attributing causes of change. This can hinder development of effective therapies due to limitations in experimental design rather than due to therapeutic failure. An improved method to model biological variation and heterogeneity in imaging signals is described. Specifically, linear Poisson modeling (LPM) evaluates changes in apparent diffusion co-efficient between baseline and 72 h after radiotherapy, in two xenograft models of colorectal cancer. The statistical significance of measured changes is compared to those attainable using a conventional t-test analysis on basic apparent diffusion co-efficient distribution parameters. Results: When LPMs were applied to treated tumors, the LPMs detected highly significant changes. The analyses were significant for all tumors, equating to a gain in power of 4-fold (i.e. equivalent to having a sample size 16 times larger), compared with the conventional approach. In contrast, highly significant changes are only detected at a cohort level using t-tests, restricting their potential use within personalized medicine and increasing the number of animals required during testing. Furthermore, LPM enabled the relative volumes of responding and non-responding tissue to be estimated for each xenograft model. Leave-one-out analysis of the treated xenografts provided quality control and identified potential outliers, raising confidence in LPM data at clinically relevant sample sizes. Availability and implementation: TINA Vision open source software is available from www.tina-vision.net. Supplementary information: Supplementary data are available at Bioinformatics online.

Quantifying biological samples using Linear Poisson Independent Component Analysis for MALDI-ToF mass spectra.

Motivation: Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI) facilitates the analysis of large organic molecules. However, the complexity of biological samples and MALDI data acquisition leads to high levels of variation, making reliable quantification of samples difficult. We present a new analysis approach that we believe is well-suited to the properties of MALDI mass spectra, based upon an Independent Component Analysis derived for Poisson sampled data. Simple analyses have been limited to studying small numbers of mass peaks, via peak ratios, which is known to be inefficient. Conventional PCA and ICA methods have also been applied, which extract correlations between any number of peaks, but we argue makes inappropriate assumptions regarding data noise, i.e. uniform and Gaussian. Results: We provide evidence that the Gaussian assumption is incorrect, motivating the need for our Poisson approach. The method is demonstrated by making proportion measurements from lipid-rich binary mixtures of lamb brain and liver, and also goat and cow milk. These allow our measurements and error predictions to be compared to ground truth. Availability and implementation: Software is available via the open source image analysis system TINA Vision, www.tina-vision.net. Contact: paul.tar@manchester.ac.uk. Supplementary information: Supplementary data are available at Bioinformatics online.

Estimating False Positive Contamination in Crater Annotations from Citizen Science Data.

Web-based citizen science often involves the classification of image features by large numbers of minimally trained volunteers, such as the identification of lunar impact craters under the Moon Zoo project. Whilst such approaches facilitate the analysis of large image data sets, the inexperience of users and ambiguity in image content can lead to contamination from false positive identifications. We give an approach, using Linear Poisson Models and image template matching, that can quantify levels of false positive contamination in citizen science Moon Zoo crater annotations. Linear Poisson Models are a form of machine learning which supports predictive error modelling and goodness-of-fits, unlike most alternative machine learning methods. The proposed supervised learning system can reduce the variability in crater counts whilst providing predictive error assessments of estimated quantities of remaining true verses false annotations. In an area of research influenced by human subjectivity, the proposed method provides a level of objectivity through the utilisation of image evidence, guided by candidate crater identifications.