Article ; Online: Digital microfluidic immunocytochemistry in single cells.
2015 Volume 6, Page(s) 7513
Abstract: We report a new technique called Digital microfluidic Immunocytochemistry in Single Cells (DISC ... DISC automates protocols for cell culture, stimulation and immunocytochemistry, enabling ... to quantitatively probe signalling events with high time resolution at the single-cell level, we propose that DISC ...
| Abstract | We report a new technique called Digital microfluidic Immunocytochemistry in Single Cells (DISC). DISC automates protocols for cell culture, stimulation and immunocytochemistry, enabling the interrogation of protein phosphorylation on pulsing with stimulus for as little as 3 s. DISC was used to probe the phosphorylation states of platelet-derived growth factor receptor (PDGFR) and the downstream signalling protein, Akt, to evaluate concentration- and time-dependent effects of stimulation. The high time resolution of the technique allowed for surprising new observations-for example, a 10 s pulse stimulus of a low concentration of PDGF is sufficient to cause >30% of adherent fibroblasts to commit to Akt activation. With the ability to quantitatively probe signalling events with high time resolution at the single-cell level, we propose that DISC may be an important new technique for a wide range of applications, especially for screening signalling responses of a heterogeneous cell population. |
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| MeSH term(s) | Animals ; Cell Culture Techniques ; Cell Line ; Fibroblasts/metabolism ; Humans ; Immunohistochemistry/instrumentation ; Immunohistochemistry/methods ; MCF-7 Cells ; Mice ; Microfluidic Analytical Techniques/instrumentation ; Microfluidic Analytical Techniques/methods ; Microfluidics ; NIH 3T3 Cells ; Phosphorylation ; Platelet-Derived Growth Factor/metabolism ; Proto-Oncogene Proteins c-akt/metabolism ; Signal Transduction |
| Chemical Substances | Platelet-Derived Growth Factor ; Proto-Oncogene Proteins c-akt (EC 2.7.11.1) |
| Language | English |
| Publishing date | 2015-06-24 |
| Publishing country | England |
| Document type | Journal Article |
| ZDB-ID | 2553671-0 |
| ISSN | 2041-1723 ; 2041-1723 |
| ISSN (online) | 2041-1723 |
| ISSN | 2041-1723 |
| DOI | 10.1038/ncomms8513 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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