Delivery of minimally dispersed liquid interfaces for sequential surface chemistry

Ostromohov, Nadya; Bercovici, Moran; Kaigala, Govind V.

doi:10.1039/c6lc00473c

Cited by 17 publications

(27 citation statements)

References 47 publications

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“…However, combining recently reported features for liquid heating 46 and liquid switching 47 in the MFP head, it is possible to implement the entire FISH protocol and automate the assay with the MFP. A shortcoming of the used device is currently the low throughput.…”

Section: Discussionmentioning

confidence: 99%

Rapid micro fluorescence in situ hybridization in tissue sections

Huber

Kaigala

2018

Biomicrofluidics

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This paper describes a micro fluorescence in situ hybridization (μFISH)-based rapid detection of cytogenetic biomarkers on formalin-fixed paraffin embedded (FFPE) tissue sections. We demonstrated this method in the context of detecting human epidermal growth factor 2 (HER2) in breast tissue sections. This method uses a non-contact microfluidic scanning probe (MFP), which localizes FISH probes at the micrometer length-scale to selected cells of the tissue section. The scanning ability of the MFP allows for a versatile implementation of FISH on tissue sections. We demonstrated the use of oligonucleotide FISH probes in ethylene carbonate-based buffer enabling rapid hybridization within <1 min for chromosome enumeration and 10–15 min for assessment of the HER2 status in FFPE sections. We further demonstrated recycling of FISH probes for multiple sequential tests using a defined volume of probes by forming hierarchical hydrodynamic flow confinements. This microscale method is compatible with the standard FISH protocols and with the Instant Quality FISH assay and reduces the FISH probe consumption ∼100-fold and the hybridization time 4-fold, resulting in an assay turnaround time of <3 h. We believe that rapid μFISH has the potential of being used in pathology workflows as a standalone method or in combination with other molecular methods for diagnostic and prognostic analysis of FFPE sections.

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Section: Discussionmentioning

confidence: 99%

Rapid micro fluorescence in situ hybridization in tissue sections

Huber

Kaigala

2018

Biomicrofluidics

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“…The apex width of 10 to 50 μm used in this study was decided based on the values in literature, e.g., 20 μm[13] and 50 μm[14]. As shown in Fig 2(d), the chemical stimulation area increased almost linearly with the flow ratio regardless of the apex width of the MFP channels.…”

Section: Resultsmentioning

confidence: 99%

“…A microfluidic probe (MFP) has been proposed for improving the spatial resolution of humoral factor stimulation control within cell culture environments[10–14]. MFPs have two microchannels, which are located adjacently across a gap of the order of tens of microns.…”

Section: Introductionmentioning

confidence: 99%

Spatial Chemical Stimulation Control in Microenvironment by Microfluidic Probe Integrated Device for Cell-Based Assay

et al. 2016

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Cell—cell interactions play an important role in the development and function of multicellular organisms. To investigate these interactions in detail, it is necessary to evaluate the behavior of a cell population when the minimum number of cells in the population is stimulated by some chemical factors. We propose a microfluidic device integrated with microfluidic probe (MFP) functionality; this device is capable of imparting a chemical stimulus to cells within a microenvironment, for cell-based assays. The device contains MFP channels at the walls of the cell culture microchannels, and it can control a localized chemical stimulation area at the scale of a single cell to a few cells using MFP fluid control in a microspace. The results of a finite element method-based simulation indicated that it is possible to control the chemical stimulation area at the scale of a single cell to a few cells by optimizing the MFP channel apex width and the flow ratio. In addition, localized cell staining was demonstrated successfully using a spatial chemical stimulus. We confirmed the device functionality as a novel cell-based assay tool. We succeeded in performing localized cell collection using this method, which suggested that the single cell analysis of a cell monolayer that is subjected to a specific chemical stimulus is possible. The method proposed in this paper can contribute significantly to the fields of cell biology and drug development.

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“…The probe head was interfaced with a syringe pump system. This allowed for manipulation of nL volumes and interrogation of an area of 300 µm × 300 µm [73,74], with low dead volumes and very short diffusion distances down to a few µm. The hybridization probe solution could also be switched over so that different areas of the sample could be interrogated with different probes.…”

Section: Vertical Microfluidic Probe For Cell Layers and Tissue Slicesmentioning

confidence: 99%

FISH and chips: a review of microfluidic platforms for FISH analysis

Rodriguez-Mateos

Azevedo

Almeida

et al. 2020

Med Microbiol Immunol

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Fluorescence in situ hybridization (FISH) allows visualization of specific nucleic acid sequences within an intact cell or a tissue section. It is based on molecular recognition between a fluorescently labeled probe that penetrates the cell membrane of a fixed but intact sample and hybridizes to a nucleic acid sequence of interest within the cell, rendering a measurable signal. FISH has been applied to, for example, gene mapping, diagnosis of chromosomal aberrations and identification of pathogens in complex samples as well as detailed studies of cellular structure and function. However, FISH protocols are complex, they comprise of many fixation, incubation and washing steps involving a range of solvents and temperatures and are, thus, generally time consuming and labor intensive. The complexity of the process, the relatively high-priced fluorescent probes and the fairly high-end microscopy needed for readout render the whole process costly and have limited wider uptake of this powerful technique. In recent years, there have been attempts to transfer FISH assay protocols onto microfluidic lab-on-a-chip platforms, which reduces the required amount of sample and reagents, shortens incubation times and, thus, time to complete the protocol, and finally has the potential for automating the process. Here, we review the wide variety of approaches for lab-on-chip-based FISH that have been demonstrated at proof-of-concept stage, ranging from FISH analysis of immobilized cell layers, and cells trapped in arrays, to FISH on tissue slices. Some researchers have aimed to develop simple devices that interface with existing equipment and workflows, whilst others have aimed to integrate the entire FISH protocol into a fully autonomous FISH on-chip system. Whilst the technical possibilities for FISH on-chip are clearly demonstrated, only a small number of approaches have so far been converted into off-the-shelf products for wider use beyond the research laboratory. Keywords Microfluidics-assisted FISH • µFISH • Microfluidics • Lab-on-a-Chip (LOC) • Fluorescence in situ hybridization (FISH) Edited by Volkhard A. J. Kempf.

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Delivery of minimally dispersed liquid interfaces for sequential surface chemistry

Cited by 17 publications

References 47 publications

Rapid micro fluorescence in situ hybridization in tissue sections

Rapid micro fluorescence in situ hybridization in tissue sections

Spatial Chemical Stimulation Control in Microenvironment by Microfluidic Probe Integrated Device for Cell-Based Assay

FISH and chips: a review of microfluidic platforms for FISH analysis

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