Cell-Based Microarrays: Review of Applications, Developments by Ella Palmer

By Ella Palmer

This booklet is a overview at the evolution of cell-based microarrays and an replace to the author's previous e-book Methods in Molecular Biology: Cell-Based Microarrays. in view that their improvement in 2001, cell-based microarrays have complicated significantly to incorporate expression arrays, brief interfering RNA arrays and antibody arrays. the outside used to coat the glass slides has additionally been considerably enhanced to permit non-adherent cells to bind to the arrays.

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By Ella Palmer

This booklet is a overview at the evolution of cell-based microarrays and an replace to the author's previous e-book Methods in Molecular Biology: Cell-Based Microarrays. in view that their improvement in 2001, cell-based microarrays have complicated significantly to incorporate expression arrays, brief interfering RNA arrays and antibody arrays. the outside used to coat the glass slides has additionally been considerably enhanced to permit non-adherent cells to bind to the arrays.

Show description

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Extended their work to both dividing and post-mitotic primary cells: Primary human BJ fibroblasts served as a representative of primary dividing cells, whereas murine bone marrow-derived dendritic cells served as a representative of nondividing cells. A higher degree of complexity was achieved in an experiment conducted on HeLa cells, which were seeded on arrays printed with lentivirus-encoding shRNA directed against lamin A/C (as a negative control) or against mammalian target of rapamycin (mTOR), a kinase involved in signal transduction cascades regulating cell size and proliferation.

The method is based on the incubation of raw sample—diluted to 1/10th in liquid culture medium according to standard methods—on a microarray and incubation at optimal temperature in an SPR imager. By monitoring in real-time the bacterial growth on the protein microarrays, remarkable LOD were reached with the detection of only a few Salmonella in less than 15 h. Interestingly, data analysis allowed the access to initial contamination levels, regardless of the strain detected in the sample. Due to the significant improvement in assay duration and easy operation, such label-free methodologies appear as promising approaches for living cell detection.

Fujita S, Ota E, Sasaki C, Takano K, Miyake M, Miyake J (2007) Highly efficient reverse transfection with siRNA in multiple wells of microtiter Plates. J Biosci Bioeng 104(4):329–333 13. Erfle H, Pepperkok R (2007) Production of siRNA- and cDNA-transfected cell arrays on noncoated chambered coverglass for high-content screening microscopy in living cells. Methods Mol Biol 360:155–161 14. Erfle H, Neumann B, Rogers P, Bulkescher J, Ellenberg J, Pepperkok R (2008) Work flow for multiplexing siRNA assays by solid-phase reverse transfection in multiwell Plates.

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