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Micro fluorescence in situ hybridization (μFISH) for spatially multiplexed analysis of a cell monolayer

HUBER1, D - Personal Name
AUTEBERT1, J - Personal Name
KAIGALA1, G. V - Personal Name

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.

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E-Jurnal

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Microfluidics
Fluorescence in situ hybridization
Microfluidic probe
Spatial multiplexing

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D. Huber1, J. Autebert1, G. V. Kaigala1

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NPP. 3509212D1000001

Micro fluorescence in situ hybridization (μFISH) for spatially multiplexed analysis of a cell monolayer

HUBER1, D - Personal NameAUTEBERT1, J - Personal NameKAIGALA1, G. V - Personal Name

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Publisher

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ISBN/ISSN

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Content Type

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Edition

Subject(s)

Specific Detail Info

Statement of Responsibility

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HUBER1, D - Personal Name
AUTEBERT1, J - Personal Name
KAIGALA1, G. V - Personal Name