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Resolution: standard / high Figure 1.
Living Microarray method. (A) Platform Workflow. The Living Microarrays method allows for highly parallelized
measurement of reporter gene expression at the single-cell level. Synthetic promoter
cassettes are cloned into the VC5 reporter construct and spotted with transfection
reagent onto glass slides. Transfected cell clusters are imaged iteratively using
automated microscopy and analyzed to generate normalized single-cell measurements
of transcriptional activity. (B) Whole chip view of a Living Microarray. HEK-293T
cells were reverse-transfected at 600 spots with varying ratios of pEGFP-N3 and Lipofectamine2000
and fixed cells were imaged at 10 μm resolution using a microarray scanner. Maximal
transfection conditions are indicated by the white box and shown at higher magnification.
(C) Reverse transfection reporter microarrays using human, mouse and rat cell lines
originating from various tissues transfected with pEGFP-N3. Scale bars = 350 μm.
Rajan et al. BMC Genomics 2011 12:115 doi:10.1186/1471-2164-12-115 |