Multi-Scale Imaging and Informatics Pipeline for In Situ Pluripotent Stem Cell Analysis

Human pluripotent stem (hPS) cells are a potential source of cells for medical therapy and an ideal system to study fate decisions in early development. However, hPS cells cultured in vitro exhibit a high degree of heterogeneity, presenting an obstacle to clinical translation. hPS cells grow in spatially patterned colony structures, necessitating quantitative single-cell image analysis. We offer a tool for analyzing the spatial population context of hPS cells that integrates automated fluorescent microscopy with an analysis pipeline. It enables high-throughput detection of colonies at low resolution, with single-cellular and sub-cellular analysis at high resolutions, generating seamless in situ maps of single-cellular data organized by colony. We demonstrate the tool's utility by analyzing inter- and intra-colony heterogeneity of hPS cell cycle regulation and pluripotency marker expression. We measured the heterogeneity within individual colonies by analyzing cell cycle as a function of distance. Cells loosely associated with the outside of the colony are more likely to be in G1, reflecting a less pluripotent state, while cells within the first pluripotent layer are more likely to be in G2, possibly reflecting a G2/M block. Our multi-scale analysis tool groups colony regions into density classes, and cells belonging to those classes have distinct distributions of pluripotency markers and respond differently to DNA damage induction. Lastly, we demonstrate that our pipeline can robustly handle high-content, high-resolution single molecular mRNA FISH data by using novel image processing techniques. Overall, the imaging informatics pipeline presented offers a novel approach to the analysis of hPS cells that includes not only single cell features but also colony wide, and more generally, multi-scale spatial configuration

Eivor Andersen Oftestad, The Lateran Church in Rome and the Ark of the Covenant: Housing the Holy Relics of Jerusalem with an Edition and Translation of the Descriptio Lateranensis Ecclesia (Bav Reg. lat. 712)

D’après la « Descriptio Lateranensis Ecclesiae », datée du xiie s. et rédigée par Jean le Diacre, l’Arche d’Alliance serait présente au palais du Latran, « in ecclesia lateranensi que est caput mundi […] et ejusdem ecclesiae principalis est archa federis domoni, vel ut aiunt archa est interius », (chap. XX, p. 222) et ce, depuis la seconde destruction du Temple par Titus, « haec autem archam cum candelabro […] tytus et vespasianus asportaverunt […] », (chap. LII, p. 223). Jean le Diacre chois..

El sistema constructivo

Se describe el sistema constructivo de entramado en madera, “plataforma” (platform frame) utilizado preferentemente en las construcciones de varios pisos en el campamento minero de Sewell, Chile./The following is a description of the platform frame used in several stories buildings in the mining settlement Sewell, in Chile

Centro-Sul

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