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1.
PLoS One ; 12(4): e0176079, 2017.
Article in English | MEDLINE | ID: mdl-28445488

ABSTRACT

Intercellular interactions play a central role at the tissue and whole organism level modulating key cellular functions in normal and disease states. Studies of cell-cell communications are challenging due to ensemble averaging effects brought about by intrinsic heterogeneity in cellular function which requires such studies to be conducted with small populations of cells. Most of the current methods for producing and studying such small cell populations are complex to implement and require skilled personnel limiting their widespread utility in biomedical research labs. We present a simple and rapid method to produce small populations with varying size of epithelial cells (10-50 cells/population) with high-throughput (~ 1 population/second) on flat surfaces via patterning of extracellular matrix (ECM) proteins and random seeding of cells. We demonstrate that despite inherent limitations of non-contact, drop-on-demand piezoelectric inkjet printing for protein patterning, varying mixtures of ECM proteins can be deposited with high reproducibility and level of control on glass substrates using a set of dynamically adjustable optimized deposition parameters. We demonstrate high consistency for the number of cells per population (~1 cell standard error of mean), the population's size (~0.2 coefficient of variation) and shape, as well as accurate spatial placement of and distance between colonies of a panel of metaplastic and dysplastic esophageal epithelial cells with differing adhesion and motility characteristics. The number of cells per colony, colony size and shape can be varied by dynamically varying the amount of ECM proteins deposited per spatial location and the number of spatial locations on the substrate. The method is applicable to a broad range of biological and biomedical studies including cell-cell communications, cellular microenvironment, migration, and stimulus response.


Subject(s)
Epithelial Cells/metabolism , Extracellular Matrix Proteins/metabolism , Cell Adhesion/physiology , Cell Shape , Cell Size , Cell Survival , Cells, Cultured , Epithelial Cells/cytology , Humans , Microscopy, Fluorescence , Substrate Specificity
2.
Nat Biotechnol ; 24(9): 1151-61, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16964229

ABSTRACT

Over the last decade, the introduction of microarray technology has had a profound impact on gene expression research. The publication of studies with dissimilar or altogether contradictory results, obtained using different microarray platforms to analyze identical RNA samples, has raised concerns about the reliability of this technology. The MicroArray Quality Control (MAQC) project was initiated to address these concerns, as well as other performance and data analysis issues. Expression data on four titration pools from two distinct reference RNA samples were generated at multiple test sites using a variety of microarray-based and alternative technology platforms. Here we describe the experimental design and probe mapping efforts behind the MAQC project. We show intraplatform consistency across test sites as well as a high level of interplatform concordance in terms of genes identified as differentially expressed. This study provides a resource that represents an important first step toward establishing a framework for the use of microarrays in clinical and regulatory settings.


Subject(s)
Gene Expression Profiling/instrumentation , Oligonucleotide Array Sequence Analysis/instrumentation , Quality Assurance, Health Care/methods , Equipment Design , Equipment Failure Analysis , Gene Expression Profiling/methods , Quality Control , Reproducibility of Results , Sensitivity and Specificity , United States
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