“How to Make It and How to Use It”, University of Kansas, Lawrence, KS August 2019

Microfluidics in Commercial Applications

Maggie A. Witek, Ph.D

Department of Chemistry

Center of BioModular Multiscale Systems for Precision Medicine

The University of Kansas

August 7-9, 2019

1

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

• The global microfluidics market size valued at $3.6 billion in 2018

• The market is expanding at a “compounded annual growth rate” (CAGR) of 18.07%.

• The market is expected to grow owing to the increasing demand of the Point-of-Care (POC) market,

supported by advances in biotechnology, including gene sequencing and in vitro diagnostics

Microfluidics Market Size

$13 billion

$3.6 billion

https://www.grandviewresearch.com/industry-analysis/microfluidics-market

2

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

Illumina, Inc.; Agilent Technologies; Caliper Life Sciences (acquired by

PerkinElmer, Inc.); Cepheid; Danaher Corporation; Life Technologies

Corporation (acquired by Thermo Fisher Scientific, Inc.); Bio-Rad

Laboratories, Inc.; Abbott Laboratories; F. Hoffmann-La Roche Ltd; and

Fluidigm Corporation.

Microfluidics Market Classification

Materialssilicone, glass, and polymer

(expected to dominate)

Componentsmicroneedles, micropumps,

microfluidic chips (largest market)

ApplicationsGenomics, Proteomics, Capillary

Electrophoresis, IVD (POC, Clinical

Diagnostics), Drug Delivery,

Microreactors, Lab Tests

Key market contributors

3

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

• Increasing preference for self-testing are driving the global point-of-care market.

• Rising incidence and prevalence of various diseases (i.e., cancer, chronic and infectious diseases).

• Decentralization of healthcare.

• Growing private investments and the availability of venture funding for the development of new products.

• Government support for improving the adoption of POC devices.

The Growth of the Microfluidics Market

https://circle.ufluidix.com/rising-demand-for-point-of-care-testing-and-significant-return-on-investment-key-driving-factors-of-the-

microfluidics-market/#more-2359

POC Devices

Pharmaceutical Industry

• Microfluidics reduces the cost per test by reducing

reagent consumption.

• Microfluidic-based micro-reactors used to control

parameters in chemical reactions and better understand

and enhance the quality of production: adopted by

Merck, Novartis, GSK, Pfizer, and Sanofi-Aventis

https://www.ddw-online.com/enabling-technologies/p149617-pharmaceutical-&-biomedical-research-likely-to-use-$1-billion-worth-of-microfluidics-devices-by-2016.html

4

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

Commercial Microfluidic Products

Trends in Biotechnology July 2014, Vol. 32, No. 7 5

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

Agilent 2100 Bioanalyzer

https://www.agilent.com/cs/library/slidepresentation/Public/Overview%20Agilent%20Microfluidics.pdf6

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

https://www.agilent.com/cs/library/posters/public/5991-8605EN_IBBL_4200_tapestation_poster.pdf

Agilent Tapestation

7

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

Next Generation Sequencing

8

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

Microarray for Hybridization of Nucleic Acids

• photolithographic process for creating DNA microarrays

• a series of photolithographic masks, light deprotection reactions, and

nucleotide coupling

• during each deprotection step, a specific mask is used with particular

transparent “windows” to allow the light from a single UV source to deprotect spots, or features on the array to receive a nucleotide. http://media.affymetrix.com/promotions/genetitan/index.affx

9

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

Droplet Generator Chip for digital PCR

QX200 Droplet Generator utilizes microfluidic chip to partition

the samples into ~20,000 nanoliter-sized droplets

10

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

• Microfluidic droplet-based platform for single cell research

• Cells encapsulated (up to 8 samples), in parallel

• Over 50,000 single cells can be captured per cartridge in a run.

A small conditional RNA (scRNA)

Nadia Instrument for Single Cell Analysis

https://www.dolomite-bio.com/product/nadia-instrument/11

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

• Applications: ddPCR, RT-PCR

• mRNA Seq: physical capture, lysis, reverse transcription and cDNA

amplification of 96 individual cells (diameter between 5 to 25 µm)

• 96 single-cell whole genome/RNA harvests for library preparation

https://www.fluidigm.com/binaries/content/documents/fluidigm/resources/c1-system-for-mrna-seq-ht-v2-pr-101-7557/c1-system-for-mrna-seq-ht-v2-pr-101-7557/fluidigm%3Afile

• Fluidigm’s integrated fluidic circuit (IFC) is based on pneumatic PDMS valves. • Fluidigm became the first company to commercialize digital PCR in 2006 (public offering in 2011).

Integrated Fluidic Circuit (IFC)

12

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

• 1 minute SDS-PAGE separation on each single-cell lysate on-chip.

• Can measure ~12 proteins per cell using a variety of strategies.

Single-Cell Western Blot Platform

13

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

• DEP cages allow trapping, manipulation, and recovery of individual cells.

• Image-based selection allows identification and isolation of cells of interest.

• The low voltage, MHz frequency impart no damage on cells.

http://www.siliconbiosystems.com/cellbrowser-software

DEPArray™

The single-use, microfluidic cartridge

contains an array of individually controllable

electrodes, each with embedded sensors.

Utilizes the ability of a non-uniform electric field to exert forces on neutral,

polarizable particles (cells) suspended in a liquid. This electrokinetic principle

is called dielectrophoresis (DEP).

When a DEP cage is moved by a change in the electric

field pattern, the trapped cell moves with it.

~40,000 DEP cages

Rare Cells Sorting

http://www.excilone.com/client/document/deparray-brochure_a4_37.pdf14

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

• Patient-side testing with the i-STAT System (POC)• Test results are uploaded automatically.

https://www.pointofcare.abbott/us/en/offerings/istat/istat-handheld

Microfluidic Blood Analyzers: i-STAT System

15

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

• The VETSCAN is a chemistry, electrolyte, immunoassay, and acid-base

analyzer from whole blood, serum, or plasma.

• Utility in veterinary clinics, mobile practitioners, research laboratories

• Initially project was NASA funded to develop a small biochemical

analyzer for use in space laboratories.

• The VETSCAN analyzer was introduced in the US veterinary

market in July 1995.

• The analyzer with the Diagnostic Profile: a nine-test reagent

rotor.

https://www.abaxis.com/page/our-history

Veterinary Diagnostics

https://www.abaxis.com/veterinary/field/biomedical-university16

“How to Make It and How to Use It”, The University of Kansas, Lawrence, KS August 2019

Conclusions

17

Chiu, et al Chem 2, 201–223, February 9, 2017

• The relevance of microfluidics remains significant

“…..our final thought is that more and more ‘‘microfluidic-embedded’’ instruments will soon be

available. However, just like your smartphone, this technology will be seamless, and all of the

engineering, chemistry, and physics that went into it will not be transparent; it will just work, and it

may very well work with your smartphone.”

https://patents.google.comhttps://scholar.google.com/

0 5 10 15 200

5000

10000

15000

20000

25000

0 5 10 15 200

2000

4000

6000

8000

10000

Nu

mb

er

of

Pu

bli

cati

on

s

Nu

mb

er

of

Pate

ntsAs of August 5, 2019 As of August,5 2019

2016

2001

2015

2019

2017

2014

2018

2010

2009

2013

2011

2008

2012

2004

2003

2007

2005

2002

2006

2000

2016

2001

2015

2019

2017

2014

2018

2010

2009

2013

2011

2008

2012

2004

2003

2007

2005

2002

2006

2000

Year Year