Deterministic encapsulation of particles and cells through viscoelastic ordering in microfluidic devices
Lead Research Organisation:
Swansea University
Department Name: College of Engineering
Abstract
New techniques which afford the rapid screening of cells to determine the presence of diseases are essential to provide efficient and effective future treatments and to improve patient health outcomes across a broad spectrum of disease states. In the last 30 years, the introduction of micro electro mechanical systems (MEMS) has prompted substantial research to develop miniaturised disease screening equipment based on microfluidic technologies. Such technologies aim to enhance point of care (POC) diagnosis, as a rapid clinical alternative to biopsy or blood tests (results typically between 2-10 days).
The encapsulation of cells together with expensive functionalised particles, called Drop-seq, is currently regarded as the most effective existing single diagnostic platform approach to derive the genome of cells affected by a variety of diseases down to the single-cell level. Although this Drop-seq technology represents the current state of the art, it suffers from a serious drawback as only approximately 10% of the cells involved are encapsulated together with a single functionalised expensive particle. This loss of over 90% of cells and expensive particles in the sensing process is a serious limitation for the screening of rare cells such as circulating tumour cells of which there are only 1/1000 in a typical blood sample. Earlier and more accurate detection of potentially fatal diseases would represent a remarkable advance in healthcare, with substantive reductions in the ongoing health cost burden and significant improvements to the quality of life of each affected individual.
The research proposed aims to exploit advances in viscoelastic flow technology to increase the efficiency of the Drop-seq technique to an unprecedented 100%. To achieve this transformative result the planned work will establish a means to ensure the equal-spacing of cells and functionalised particles before they approach the encapsulation area. This ensures that a single cell is encapsulated with a single particle, in a single droplet (a process referred to herein as deterministic encapsulation). Thus, the Drop-seq becomes deterministic when the frequency of droplet formation is synchronised with the frequency of particles and cells approaching the encapsulation area. Henceforth, the efficiency of the Drop-seq technology becomes 100%, rather than the mere 10% currently obtained.
The project will be carried out in conjunction with an industry partner that is recognised as the world leader in the design and manufacture of pioneering microfluidic products.
The objectives of the research are as follows:
(1) Achieving cell ordering in straight channel; (2) Achieving continuous formation of viscoelastic droplets with uniform sizes and shapes; (3) Encapsulation of single particles in viscoelastic droplets with uniform size distribution; (4) Encapsulation of single cells in viscoelastic droplets.
The research, which will achieve unprecedented disease detection capability, has substantive potential impacts, both in terms of healthcare outcomes and economic benefits. It will provide a basis for transforming the accuracy of detection for diseases such as lung, prostate, breast and bowel cancers, which currently account for more than half the types of cancer and lead to premature death. Beneficiaries will therefore include patients and healthcare professionals. Specifically, the POC testing approach has special relevance to healthcare professional working in remote locations. Examples include GP surgeries, clinics in remote locations or even within pharmacies. Our project partner is well placed to drive the research outputs to commercialisation and economic gain- on a global basis. Furthermore, widespread technical benefits in diverse fields including Process Engineering and Advanced Manufacturing may be anticipated to arise from enhanced knowledge of the innovative uses of viscoelastic flows within microfluidic settings.
The encapsulation of cells together with expensive functionalised particles, called Drop-seq, is currently regarded as the most effective existing single diagnostic platform approach to derive the genome of cells affected by a variety of diseases down to the single-cell level. Although this Drop-seq technology represents the current state of the art, it suffers from a serious drawback as only approximately 10% of the cells involved are encapsulated together with a single functionalised expensive particle. This loss of over 90% of cells and expensive particles in the sensing process is a serious limitation for the screening of rare cells such as circulating tumour cells of which there are only 1/1000 in a typical blood sample. Earlier and more accurate detection of potentially fatal diseases would represent a remarkable advance in healthcare, with substantive reductions in the ongoing health cost burden and significant improvements to the quality of life of each affected individual.
The research proposed aims to exploit advances in viscoelastic flow technology to increase the efficiency of the Drop-seq technique to an unprecedented 100%. To achieve this transformative result the planned work will establish a means to ensure the equal-spacing of cells and functionalised particles before they approach the encapsulation area. This ensures that a single cell is encapsulated with a single particle, in a single droplet (a process referred to herein as deterministic encapsulation). Thus, the Drop-seq becomes deterministic when the frequency of droplet formation is synchronised with the frequency of particles and cells approaching the encapsulation area. Henceforth, the efficiency of the Drop-seq technology becomes 100%, rather than the mere 10% currently obtained.
The project will be carried out in conjunction with an industry partner that is recognised as the world leader in the design and manufacture of pioneering microfluidic products.
The objectives of the research are as follows:
(1) Achieving cell ordering in straight channel; (2) Achieving continuous formation of viscoelastic droplets with uniform sizes and shapes; (3) Encapsulation of single particles in viscoelastic droplets with uniform size distribution; (4) Encapsulation of single cells in viscoelastic droplets.
The research, which will achieve unprecedented disease detection capability, has substantive potential impacts, both in terms of healthcare outcomes and economic benefits. It will provide a basis for transforming the accuracy of detection for diseases such as lung, prostate, breast and bowel cancers, which currently account for more than half the types of cancer and lead to premature death. Beneficiaries will therefore include patients and healthcare professionals. Specifically, the POC testing approach has special relevance to healthcare professional working in remote locations. Examples include GP surgeries, clinics in remote locations or even within pharmacies. Our project partner is well placed to drive the research outputs to commercialisation and economic gain- on a global basis. Furthermore, widespread technical benefits in diverse fields including Process Engineering and Advanced Manufacturing may be anticipated to arise from enhanced knowledge of the innovative uses of viscoelastic flows within microfluidic settings.
Planned Impact
The project targets the establishment of a viable microfluidic prototype for point-of-care (POC) applications based on the deterministic encapsulation of particles and cells suspended in viscoelastic polymer solution flowing in microfluidic devices. The prototype will be developed and commercialised in conjunction with an industry partner that is recognised as the World leader in the design and manufacture of pioneering microfluidic products.
Health Impact -The eventual availability of a new healthcare product will orchestrate marked benefits for patients and healthcare professionals. Early diagnosis of disease is critical to achieve effective patient care and improve health outcomes. Obtaining a blood sample takes several minutes, blood analysis can take significantly longer. To expedite clinical tests, for point-of-care, tools based on microfluidic devices have been developed. These include so called 'lab-on-a-chip' devices that analyse blood in relation to common clinical parameters including white blood cell count, platelet count and haemoglobin.
The proposed research will extend existing capabilities and significantly improve the analysis of single cells with lab-on-a-chip technologies, allowing highly efficient screening of blood diseases (analysing both white and red blood cells), for a wider range of disorders. For instance, the encapsulation of red blood cells with functionalised particles can be used for the genome identification of the cells, allowing the screening of a variety of gene-related diseases (Drop-seq technology). Notably, currently commercialised products offer an efficiency of only 10%. The presented research will establish a novel technique based on the use of polymer solutions to boost the efficiency close to 100% - resulting in a superior commercialised product.
Patients will benefit from very detailed diagnosis of diseases (single-cell analysis). Healthcare professionals will be advantaged by the ability to deliver more accurate and efficient point-of-care duties. The product could be utilised by Health Boards (UK and international) and will be pertinent to medicine in unconventional or remote locations - military bases; field hospitals, submarines, ships; oil and gas rigs; rural healthcare centres and international Embassies.
Economic Impact -The development and commercialisation of the prototype microfluidic device will realise significant financial benefits for the industry partner, as it will financially gain from a new product with unprecedented efficiency. Subsequently, the industry partner will profit from the (probable) high sales figures associated with widespread demand for a technically superior diagnostic device.
Other potential financial beneficiaries are end users - i.e NHS; Government bodies, responsible for military medicine - who will be advantaged from a technically advanced device in comparison to less accurate and less flexible alternative existing products.
Public Engagement -The use of chemical engineering to create a new healthcare product, involving the innovative application of polymer solutions (typically associated with plastics and negative environmental impact) promises to stimulate public interest and lead to knowledge enhancement. Beneficiaries include School children (supporting an interest in STEM); healthcare support groups and patient support organisations.
Health Impact -The eventual availability of a new healthcare product will orchestrate marked benefits for patients and healthcare professionals. Early diagnosis of disease is critical to achieve effective patient care and improve health outcomes. Obtaining a blood sample takes several minutes, blood analysis can take significantly longer. To expedite clinical tests, for point-of-care, tools based on microfluidic devices have been developed. These include so called 'lab-on-a-chip' devices that analyse blood in relation to common clinical parameters including white blood cell count, platelet count and haemoglobin.
The proposed research will extend existing capabilities and significantly improve the analysis of single cells with lab-on-a-chip technologies, allowing highly efficient screening of blood diseases (analysing both white and red blood cells), for a wider range of disorders. For instance, the encapsulation of red blood cells with functionalised particles can be used for the genome identification of the cells, allowing the screening of a variety of gene-related diseases (Drop-seq technology). Notably, currently commercialised products offer an efficiency of only 10%. The presented research will establish a novel technique based on the use of polymer solutions to boost the efficiency close to 100% - resulting in a superior commercialised product.
Patients will benefit from very detailed diagnosis of diseases (single-cell analysis). Healthcare professionals will be advantaged by the ability to deliver more accurate and efficient point-of-care duties. The product could be utilised by Health Boards (UK and international) and will be pertinent to medicine in unconventional or remote locations - military bases; field hospitals, submarines, ships; oil and gas rigs; rural healthcare centres and international Embassies.
Economic Impact -The development and commercialisation of the prototype microfluidic device will realise significant financial benefits for the industry partner, as it will financially gain from a new product with unprecedented efficiency. Subsequently, the industry partner will profit from the (probable) high sales figures associated with widespread demand for a technically superior diagnostic device.
Other potential financial beneficiaries are end users - i.e NHS; Government bodies, responsible for military medicine - who will be advantaged from a technically advanced device in comparison to less accurate and less flexible alternative existing products.
Public Engagement -The use of chemical engineering to create a new healthcare product, involving the innovative application of polymer solutions (typically associated with plastics and negative environmental impact) promises to stimulate public interest and lead to knowledge enhancement. Beneficiaries include School children (supporting an interest in STEM); healthcare support groups and patient support organisations.
Publications
Shahrivar K
(2022)
Beating Poisson stochastic particle encapsulation in flow-focusing microfluidic devices using viscoelastic liquids
in Soft Matter
Shahrivar K
(2021)
Controlled viscoelastic particle encapsulation in microfluidic devices.
in Soft matter
Maisto A
(2022)
Continuous Manufacturing of Microfluidic Fibers Embedded with Ordered Microparticles via Ionic Gelation
in ACS Applied Engineering Materials
Keshvad Shahrivar
(2020)
Generation and dynamics of xanthan gum viscoelastic droplets in a T-junction microchannel
Jeyasountharan A
(2022)
Confinement effect on the viscoelastic particle ordering in microfluidic flows: Numerical simulations and experiments
in Physics of Fluids
Jeyasountharan A
(2021)
Viscoelastic Particle Train Formation in Microfluidic Flows Using a Xanthan Gum Aqueous Solution.
in Analytical chemistry
Jeyasountharan A
(2023)
Viscoelastic Particle Encapsulation Using a Hyaluronic Acid Solution in a T-Junction Microfluidic Device.
in Micromachines
Francesco Del Giudice
(2022)
Space-time evolution of microfluidic crystals from dilute viscoelastic suspensions
Francesco Del Giudice
(2020)
Simultaneous measurement of rheological properties in a microfluidic rheometer
Faroughi S
(2022)
Microfluidic Rheometry and Particle Settling: Characterizing the Effect of Polymer Solution Elasticity
in Polymers
Del Giudice F
(2022)
A Review of Microfluidic Devices for Rheological Characterisation.
in Micromachines
Del Giudice F
(2022)
Microfluidic rheometry for the rheological characterisation of polymer solution
Del Giudice F
(2022)
Rapid Temperature-Dependent Rheological Measurements of Non-Newtonian Solutions Using a Machine-Learning Aided Microfluidic Rheometer.
in Analytical chemistry
Del Giudice F
(2020)
Simultaneous measurement of rheological properties in a microfluidic rheometer
in Physics of Fluids
Del Giudice F
(2021)
Microfluidic formation of crystal-like structures.
in Lab on a chip
Barnes C
(2023)
Machine learning enhanced droplet microfluidics
in Physics of Fluids
Anoshanth Jeyasountharan
(2020)
Shear-thinning induced particle train formation in microfluidic flows
Description | The overarching goal of this proposal was to exploit the recently observed phenomenon of formation of equally-spaced particles in microfluidic flow when the suspending liquid was a viscoelastic solution. We obtained several key results, detailed below: Elucidation of the particle ordering phenomenon - We performed several experiments to understand the impact of fluid rheology and particle size on the formation of equally-spaced particles in microfluidic devices. Our experiments were also supported by numerical simulations carried out by our collaborators. We also introduced a microfluidic device to reduce the occurrence of particle aggregates. Controlled Encapsulation of particles - We demonstrated controlled encapsulation and co-encapsulation of particles in microfluidic droplets with encapsulation efficiency up to 2-folds larger compared to the stochastic value. We demonstrated controlled encapsulation in both T-junction ad Flow-focusing geometries, while we demonstrated co-encapsulation in flow-focusing geometries. Fabrication of a new class of materials - We coupled the particle ordering phenomena to the well-established procedure for fibre synthesis to manufacture a new class of materials featuring fibres with equally-spaced particles suspended in them. |
Exploitation Route | Our results are all aligned with the Healthcare theme. The possibility of achieving equally-spaced structures in microfluidic devices has the potential to solve recognition and sorting problems in flow cytometry, linked to the fact that cells arrive not equally-spaced to the sorting area. Similarly, our methodology to encapsulate objects with efficiency far above the stochastic limit is crucial in single-cell analysis and for the screening of circulating tumour cells. The formation of fibres with equally-spaced structures can be employed to manufacture a new class of sensors, while also being essential in drug delivery research and in the understanding of cell-to-cell interactions when subjected to external stimuli. |
Sectors | Healthcare,Manufacturing, including Industrial Biotechology |
Description | The finding have been used in two areas: i) knowledge exchange with the industrial partner on the proposal (Dolomite Microfluidics) for future commercialisation of the grant results; and ii) Outreach activities for the general public. With reference to Point 1, we filed a patent-pending application, PCT/EP2022/059519, that the Indutrial partner of the project want to commercialised under exclusive rights (we have a letter of intent in this respect. This entry is linked to an economical impact for the company that would embed along its supply chain the unique methodology developed under this grant. With reference to Point 2, I participated in the Swansea Science Festival where I prepared an interactive experiments where families were invited to produce droplets by changing the volumetric flow rate of two non-miscible flow phases in a flow-focusing microfluidic device. The impact was an appreciation of the importance of droplets together with the fact that these can be produced using very small devices rather than large industrial plants. |
First Year Of Impact | 2022 |
Sector | Education,Healthcare,Pharmaceuticals and Medical Biotechnology |
Impact Types | Societal,Economic |
Description | A novel holistic approach to Welsh heritage: a bilingual journey from Science and Literature to Art. |
Amount | £5,050 (GBP) |
Organisation | Welsh Crucible |
Sector | Public |
Country | United Kingdom |
Start | 01/2022 |
End | 10/2022 |
Description | Dynamics and stability of viscoelastic phonons in microfluidic devices |
Amount | £17,708 (GBP) |
Funding ID | RGS\R1\221263 |
Organisation | The Royal Society |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 03/2022 |
End | 09/2023 |
Description | IMPACT Equipment grant |
Amount | £30,000 (GBP) |
Organisation | Swansea University |
Sector | Academic/University |
Country | United Kingdom |
Start | 03/2021 |
Description | Sêr Cymru enhancing competitiveness equipment award 2022 to 2023 |
Amount | £104,235 (GBP) |
Organisation | Government of Wales |
Sector | Public |
Country | United Kingdom |
Start | 01/2023 |
End | 03/2023 |
Description | Syringe grant |
Amount | £800 (GBP) |
Organisation | Hamilton Company |
Sector | Private |
Country | United States |
Start | 01/2022 |
Description | ValidaOon of new rheological biomarkers for diagnosis and monitoring of cardiovascular diseases using a novel microfluidic technology. |
Amount | £15,000 (GBP) |
Organisation | Engineering and Physical Sciences Research Council (EPSRC) |
Department | Impact Accelerator Award |
Sector | Public |
Country | United Kingdom |
Start | 07/2023 |
End | 04/2024 |
Description | WDNA Super Sprint Projects |
Amount | £125,000 (GBP) |
Organisation | European Social Fund (Welsh Government/ EU) |
Sector | Public |
Country | United Kingdom |
Start | 02/2022 |
End | 03/2022 |
Description | µRIFLEX: Microfluidic rheometry to study RIgid-to-FLEXible polymer transition in dilute polyelectrolyte solutions |
Amount | £5,473 (GBP) |
Funding ID | IES\R2\202142 |
Organisation | The Royal Society |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 12/2020 |
End | 11/2022 |
Description | A novel holistic approach to Welsh heritage: a bilingual journey from Science and Literature to Art. |
Organisation | Cardiff University |
Department | School of Welsh |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | I contributed with my know-how in chemical engineering. |
Collaborator Contribution | Cardiff University brought experience with Welsh literature, while the University of South Wales brought experience in performative art. |
Impact | Welsh Crucible grant - A novel holistic approach to Welsh heritage: a bilingual journey from Science and Literature to Art. (~£6k) |
Start Year | 2022 |
Description | A novel holistic approach to Welsh heritage: a bilingual journey from Science and Literature to Art. |
Organisation | University of South Wales |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | I contributed with my know-how in chemical engineering. |
Collaborator Contribution | Cardiff University brought experience with Welsh literature, while the University of South Wales brought experience in performative art. |
Impact | Welsh Crucible grant - A novel holistic approach to Welsh heritage: a bilingual journey from Science and Literature to Art. (~£6k) |
Start Year | 2022 |
Description | Machine learning and AI-aided microfluidic applications |
Organisation | Swansea University |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | I provided the experimental and theoretical know-how for the characterisation of polymer solutions using microfluidic techniques. I also brought the experimental and theoretical know-how regarding droplet formation in microfluidics |
Collaborator Contribution | Experience in developing machine learning algorithms. |
Impact | Publication - Rapid Temperature-Dependent Rheological Measurements of Non- Newtonian Solutions Using a Machine-Learning Aided Microfluidic Rheometer IMPACT equipment grant - ~£30k WDNA Sprint Project - ~£125k, Intelligent Microfluidic Droplet Generator |
Start Year | 2021 |
Description | µRIFLEX: Microfluidic rheometry to study RIgid-to-FLEXible polymer transition in dilute polyelectrolyte solutions |
Organisation | RWTH Aachen University |
Country | Germany |
Sector | Academic/University |
PI Contribution | Providing the experimental know-how to perform rheological measurements of shear viscosity, longest relaxation time and storage modulus on polyelectrolyte solutions. The measurement of the longest relaxation time was not possible via conventional bulk techniques. |
Collaborator Contribution | The partner brought the know-how regarding the synthesis and chemistry of polyelectrolyte solutions as a function of added salt. |
Impact | Royal Society International Exchange Grant, #IES\R2\202142 |
Start Year | 2021 |
Title | PARTICLE SEPARATION SYSTEMS AND METHODS |
Description | The invention relates to methods for separating particles in a microfluidic device and, ideally, encapsulating said particles in at least one or a stream of droplets; and a kit of parts for performing said methods. |
IP Reference | PCT/EP2022/059519 |
Protection | Patent / Patent application |
Year Protection Granted | 2021 |
Licensed | No |
Impact | The industrial partner on the project has agreed to license the discovery by providing a letter of intent |
Title | uRheo |
Description | Software to evaluate rheological properties via analysis of particle flowing in microchannels. |
Type Of Technology | Webtool/Application |
Year Produced | 2022 |
Open Source License? | Yes |
Impact | Rheological characterisation of polymer solutions could be realised in less than 2 minutes using fingerprick of sample. |
Description | Open Days at Swansea University |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | 50 to 100 students attended the open days in 2021 and 2022, where they learnt about chemical engineering and the research carried out at Swansea University. |
Year(s) Of Engagement Activity | 2021,2022 |
Description | Outreach activities in Chemical Engineering |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Other audiences |
Results and Impact | I did organise several outreach activities with different groups of schools to introduce them to the world of chemical engineering. |
Year(s) Of Engagement Activity | 2022,2023 |