Immunofluorescence staining of paraffin sections: creating DAB staining like virtual digital images using CMYK color conversion

Crystal violet treatment of formalin fixed paraffin embedded tissue slides greatly reduces the endogenous autofluorescence, and allows immunofluorescence (IF) staining with FITC or Alexa488 conjugated antibodies. Using cold CCD camera to capture the fluorescence images makes this staining method very sensitive. Here we show that combination of IF with the simultaneous recording of crystal violet induced red and Hoechst 33258 induced blue fluorescence permits the localization of the IF signal over a cytoplasmic: nuclear red:blue stain that visualizes the microscopic anatomy of the underlying tissue. To make the visual interpretation of the IF staining easier for microscopists, who are used to DAB staining over weak hematoxilin-eosin background, we created a simple color conversion procedure that turns the captured three-color fluorescence RGB (red, green, blue) images over a black background into four color CMYK (cyan, magenta, yellow, key color (black)) images.Окраска кристаллическим фиолетовым парафиновых срезов, полученных из ткани, фиксированной в формалине, значительно уменьшает явления аутофлуоресценции и обеспечивает иммунофлуоресцентную (ИФ) окраску антителами, конъюгированными сFITC или Aleхa-488. Использование CCD-камер для регистрации флуоресцентных изображений делает этот метод очень чувствительным. Наша цель — разработать метод трансформации RGB (red, green, blue) — флуоресцентных изображений в CMYC (cyan, magenta, yellow, key color (black) ) изображения. Показано, что комбинация ИФ с одновременной регистрацией красной и голубой флуоресценции, индуцированной соответственно кристаллическим фиолетовым и Hoechst33258, позволяет определять ИФ-сигнал как цитоплазматично-ядерное красно-голубое окрашивание, которое визуализирует морфологические особенности прилегающей ткани. Для упрощения интерпретации ИФ-окраски патологами, привыкшими к окрашиванию ДАБ на фоне гематоксилин-эозина, нами была создана технология простого цветового перехода, который превращает зарегистрированные трехцветные RGB-флуоресцентные изображения на черном фоне в четырехцветные CMYC-изображения на белом фоне, используя программы работы с изображениями

A Test of Independence in Two-Way Contingency Tables Based on Maximal Correlation

Cataloged from PDF version of article.Maximal correlation has several desirable properties as a measure of dependence, including the fact that it vanishes if and only if the variables are independent. Except for a few special cases, it is hard to evaluate maximal correlation explicitly. We focus on two-dimensional contingency tables and discuss a procedure for estimating maximal correlation, which we use for constructing a test of independence. We compare the maximal correlation test with other tests of independence by Monte Carlo simulations. When the underlying continuous variables are dependent but uncorrelated, we point out some cases for which the new test is more powerful

Differential Functional Connectivity of Rostral Anterior Cingulate Cortex During Emotional Interference

The rostral-ventral subdivision of the anterior cingulate cortex (rACC) plays a key role in the regulation of emotional processing. Although rACC has strong anatomical connections with anterior insular cortex (AIC), amygdala, prefrontal cortex and striatal brain regions, it is unclear whether the functional connectivity of rACC with these regions changes when regulating emotional processing. Furthermore, it is not known whether this connectivity changes with deficits in emotion regulation seen in different kinds of anxiety and depression. To address these questions regarding rACC functional connectivity, non-patients high in self-reported anxious apprehension (AP), anxious arousal (AR), anhedonic depression (AD) or none (CON) indicated the ink color of pleasant, neutral and unpleasant words during functional magnetic resonance imaging. While ignoring task-irrelevant unpleasant words, AD and CON showed an increase in the functional connectivity of rACC with AIC, putamen, caudate and ventral pallidum. There was a decrease in this connectivity in AP and AR, with AP showing greater reduction than AR. These findings provide support for the role of rACC in integrating interoceptive, emotional and cognitive functions via interactions with insula and striatal regions during effective emotion regulation in healthy individuals and a failure of this integration that may be specific to anxiety, particularly AP

Activity of 50 Long-Period Comets Beyond 5.2 AU

Remote investigations of the ancient solar system matter has been traditionally carried out through the observations of long-period (LP) comets that are less affected by solar irradiation than the short-period counterparts orbiting much closer to the Sun. Here we summarize the results of our decade-long survey of the distant activity of LP comets. We found that the most important separation in the dataset is based on the dynamical nature of the objects. Dynamically new comets are characterized by a higher level of activity on average: the most active new comets in our sample can be characterized by afrho values >3--4 higher than that of our most active returning comets. New comets develop more symmetric comae, suggesting a generally isotropic outflow. Contrary to this, the coma of recurrent comets can be less symmetrical, ocassionally exhibiting negative slope parameters, suggesting sudden variations in matter production. The morphological appearance of the observed comets is rather diverse. A surprisingly large fraction of the comets have long, teniouos tails, but the presence of impressive tails does not show a clear correlation with the brightness of the comets.Comment: 21 pages, 4 figures, accepted for publication in A

Differential Functional Connectivity of Rostral Anterior Cingulate Cortex During Emotional Interference

The rostral-ventral subdivision of the anterior cingulate cortex (rACC) plays a key role in the regulation of emotional processing. Although rACC has strong anatomical connections with anterior insular cortex (AIC), amygdala, prefrontal cortex and striatal brain regions, it is unclear whether the functional connectivity of rACC with these regions changes when regulating emotional processing. Furthermore, it is not known whether this connectivity changes with deficits in emotion regulation seen in different kinds of anxiety and depression. To address these questions regarding rACC functional connectivity, non-patients high in self-reported anxious apprehension (AP), anxious arousal (AR), anhedonic depression (AD) or none (CON) indicated the ink color of pleasant, neutral and unpleasant words during functional magnetic resonance imaging. While ignoring task-irrelevant unpleasant words, AD and CON showed an increase in the functional connectivity of rACC with AIC, putamen, caudate and ventral pallidum. There was a decrease in this connectivity in AP and AR, with AP showing greater reduction than AR. These findings provide support for the role of rACC in integrating interoceptive, emotional and cognitive functions via interactions with insula and striatal regions during effective emotion regulation in healthy individuals and a failure of this integration that may be specific to anxiety, particularly AP

Genetic associations with childhood brain growth, defined in two longitudinal cohorts

Genome-wide association studies (GWASs) are unraveling the genetics of adult brain neuroanatomy as measured by cross-sectional anatomic magnetic resonance imaging (aMRI). However, the genetic mechanisms that shape childhood brain development are, as yet, largely unexplored. In this study we identify common genetic variants associated with childhood brain development as defined by longitudinal aMRI. Genome-wide single nucleotide polymorphism (SNP) data were determined in two cohorts: one enriched for attention-deficit/hyperactivity disorder (ADHD) (LONG cohort: 458 participants; 119 with ADHD) and the other from a population-based cohort (Generation R: 257 participants). The growth of the brain's major regions (cerebral cortex, white matter, basal ganglia, and cerebellum) and one region of interest (the right lateral prefrontal cortex) were defined on all individuals from two aMRIs, and a GWAS and a pathway analysis were performed. In addition, association between polygenic risk for ADHD and brain growth was determined for the LONG cohort. For white matter growth, GWAS meta-analysis identified a genome-wide significant intergenic SNP (rs12386571, P = 9.09 × 10-9 ), near AKR1B10. This gene is part of the aldo-keto reductase superfamily and shows neural expression. No enrichment of neural pathways was detected and polygenic risk for ADHD was not associated with the brain growth phenotypes in the LONG cohort that was enriched for the diagnosis of ADHD. The study illustrates the use of a novel brain growth phenotype defined in vivo for further study