Micro fluorescence in situ hybridization (μFISH) for spatially multiplexed analysis of a cell monolayer.
We here present a micrometer-scale implementation of fluorescence in situ hybridization that we term μFISH. This μFISH implementation makes use of a non-contact scanning probe technology, namely, a microfluidic probe (MFP) that hydrodynamically shapes nanoliter volumes of liquid on a surface with mi...
| Publicado en: | Biomedical Microdevices Vol. 18; no. 2; pp. 40 - 48 |
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| Autores principales: | , , |
| Formato: | Journal Article |
| Publicado: |
Springer Nature
Apr2016
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=115165208&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 115165208 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 13872176 ODN jtl: Biomedical Microdevices issn: 13872176 maglogo: N pubinfo: dt: Apr2016 vid: 18 iid: 2 pid: 237 pub: Springer Nature place: New York, New York artinfo: ui: 115165208 10.1007/s10544-016-0064-0 115165208 ppf: 40 ppct: 8 formats: fmt: @attributes: type: P tig: atl: Micro fluorescence in situ hybridization (μFISH) for spatially multiplexed analysis of a cell monolayer. aug: au: Huber, D. Autebert, J. Kaigala, G. V. affil: IBM Research – Zurich, Säumerstrasse 4, 8803 Rüschlikon, Switzerland. sug: ab: We here present a micrometer-scale implementation of fluorescence in situ hybridization that we term μFISH. This μFISH implementation makes use of a non-contact scanning probe technology, namely, a microfluidic probe (MFP) that hydrodynamically shapes nanoliter volumes of liquid on a surface with micrometer resolution. By confining FISH probes at the tip of this microfabricated scanning probe, we locally exposed approximately 1000 selected MCF-7 cells of a monolayer to perform incubation of probes—the rate-limiting step in conventional FISH. This method is compatible with the standard workflow of conventional FISH, allows rebudgeting of the sample for various tests, and results in a ~ 15-fold reduction in probe consumption. The continuous flow of probes and shaping liquid on these selected cells resulted in a 120-fold reduction of the hybridization time compared with the standard protocol (3 min vs. 6 h) and efficient rinsing, thereby shortening the total FISH assay time for centromeric probes.We further demonstrated spatially multiplexed μFISH, enabling the use of spectrally equivalent probes for detailed and real-time analysis of a cell monolayer, which paves the way towards rapid and automated multiplexed FISH on standard cytological supports. pubtype: Academic Journal doctype: Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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