Optimizing Staining Protocols for Laser Microdissection of Specific Cell Types from the Testis Including Carcinoma In Situ

Microarray and RT-PCR based methods are important tools for analysis of gene expression; however, in tissues containing many different cells types, such as the testis, characterization of gene expression in specific cell types can be severely hampered by noise from other cells. The laser microdissection technology allows for enrichment of specific cell types. However, when the cells are not morphologically distinguishable, it is necessary to use a specific staining method for the target cells. In this study we have tested different fixatives, storage conditions for frozen sections and staining protocols, and present two staining protocols for frozen sections, one for fast and specific staining of fetal germ cells, testicular carcinoma in situ cells, and other cells with embryonic stem cell-like properties that express the alkaline phosphatase, and one for specific staining of lipid droplet-containing cells, which is useful for isolation of the androgen-producing Leydig cells. Both protocols retain a morphology that is compatible with laser microdissection and yield RNA of a quality suitable for PCR and microarray analysis

Stochastic particle packing with specified granulometry and porosity

This work presents a technique for particle size generation and placement in arbitrary closed domains. Its main application is the simulation of granular media described by disks. Particle size generation is based on the statistical analysis of granulometric curves which are used as empirical cumulative distribution functions to sample from mixtures of uniform distributions. The desired porosity is attained by selecting a certain number of particles, and their placement is performed by a stochastic point process. We present an application analyzing different types of sand and clay, where we model the grain size with the gamma, lognormal, Weibull and hyperbolic distributions. The parameters from the resulting best fit are used to generate samples from the theoretical distribution, which are used for filling a finite-size area with non-overlapping disks deployed by a Simple Sequential Inhibition stochastic point process. Such filled areas are relevant as plausible inputs for assessing Discrete Element Method and similar techniques

Children’s Initiatives in the Finnish Early Childhood Education Context

Pedagogical practices in early childhood education, which embrace children’s initiatives and agency, have been found to have an effect on children’s learning and competence skills. These initiatives can be seen crucial for children’s well-being and self-motivation. However, children’s initiatives are sometimes considered only as wants and that children are incapable to express meaningful initiatives in educational settings. In this chapter, we introduce children’s initiatives in their educational settings and examine the gap between children’s experiences and teachers’ observations. Children’s initiatives exist in a myriad of ways through the daily practices and processes that nourish motivation to ultimately create meanings through actions. It is essential to focus on children’s participation as it encourages and promotes agency and motivation within early childhood development.Peer reviewe

A922 Sequential measurement of 1 hour creatinine clearance (1-CRCL) in critically ill patients at risk of acute kidney injury (AKI)

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