Dataset Information

Circulating tumour cell RNA characterisation from colorectal cancer patient blood after inertial microfluidic enrichment.

ABSTRACT: The detection and molecular analysis of circulating tumour cells (CTCs) potentially provides a significant insight to the characterisation of disease, stage of progression and therapeutic options for cancer patients. Following on from the protocol by Warkiani et al. 2016, which describes a method of enriching CTCs from cancer patient blood with inertial microfluidics, we describe a method to measure the CTC RNA expression in the enriched fraction using droplet digital PCR and compare transcript detection with and without RNA pre-amplification. •Inertial microfluidics combined with droplet digital PCR is advantageous as it allows for CTC enrichment and subsequent RNA analysis from patient blood. This allows for patient tumour analysis with increased sensitivity and precision compared to quantitative Real Time PCR and enables the direct quantification of nucleic acids without the need for tumour biopsy.•This method demonstrates an efficient approach providing important insights into the analysis of colorectal cancer patients' CTCs using a specific gene subset or biomarkers, an approach that may be tailored to tumour type or expanded to larger panels.

SUBMITTER: Winter M

PROVIDER: S-EPMC6603295 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Publications

Circulating tumour cell RNA characterisation from colorectal cancer patient blood after inertial microfluidic enrichment.

Winter Marnie M Cai Zhen Z Winkler Katharina K Georgiou Kristen K Inglis Daniel D Lavranos Tina T Rezaei Meysam M Warkiani Majid M Thierry Benjamin B

MethodsX 20190617

The detection and molecular analysis of circulating tumour cells (CTCs) potentially provides a significant insight to the characterisation of disease, stage of progression and therapeutic options for cancer patients. Following on from the protocol by Warkiani et al. 2016, which describes a method of enriching CTCs from cancer patient blood with inertial microfluidics, we describe a method to measure the CTC RNA expression in the enriched fraction using droplet digital PCR and compare transcript ...[more]

PMID: 31304099

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Project description:Inertial microfluidics has been broadly investigated, resulting in the development of various applications, mainly for particle or cell separation. Lateral migrations of these particles within a microchannel strictly depend on the channel design and its cross-section. Nonetheless, the fabrication of these microchannels is a continuous challenging issue for the microfluidic community, where the most studied channel cross-sections are limited to only rectangular and more recently trapezoidal microchannels. As a result, a huge amount of potential remains intact for other geometries with cross-sections difficult to fabricate with standard microfabrication techniques. In this study, by leveraging on benefits of additive manufacturing, we have proposed a new method for the fabrication of inertial microfluidic devices. In our proposed workflow, parts are first printed via a high-resolution DLP/SLA 3D printer and then bonded to a transparent PMMA sheet using a double-coated pressure-sensitive adhesive tape. Using this method, we have fabricated and tested a plethora of existing inertial microfluidic devices, whether in a single or multiplexed manner, such as straight, spiral, serpentine, curvilinear, and contraction-expansion arrays. Our characterizations using both particles and cells revealed that the produced chips could withstand a pressure up to 150?psi with minimum interference of the tape to the total functionality of the device and viability of cells. As a showcase of the versatility of our method, we have proposed a new spiral microchannel with right-angled triangular cross-section which is technically impossible to fabricate using the standard lithography. We are of the opinion that the method proposed in this study will open the door for more complex geometries with the bespoke passive internal flow. Furthermore, the proposed fabrication workflow can be adopted at the production level, enabling large-scale manufacturing of inertial microfluidic devices.

Dataset Information

Circulating tumour cell RNA characterisation from colorectal cancer patient blood after inertial microfluidic enrichment.

Publications

Circulating tumour cell RNA characterisation from colorectal cancer patient blood after inertial microfluidic enrichment.

Similar Datasets

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