µfluidics: a toolbox for analysis automation and … › cluster › event › event › 20121004...

Guillaume DelapierreCEA Leti‐Minatec / DTBS

[email protected]

+33 4 38 78 23 45

µFluidics: a ToolBox forAnalysis Automation and

Delocalization

© CEA. All rights reserved

Biology and Microfluidic Architecture Laboratory – G. Delapierre – Leti Day in Nagoya, October 4th 2012 | 2

LETI Health 2012 FiguresAround 20 M€ budget per year

133 permanent people

57 young non permanent researchers

2500 m² of lab space

500 m² of clean room

A dedicated chemistry platform

290 patent portfolio (40/year)

80 publications/year

4 common labs with industrial partners

5 start‐ups



LETI Health

Microfluidics

Instrumentation Mechanics SoftwarePackaging

Chemistry Biochemistry

Optics / Photonics

Biology

Microtechnologies

Detection

Electronics

Information processing

Electrochemistry

Modeling

Multidisciplinary teamsOne stop shop for completely integrated systems

Basic ResearchPublications

Applied ResearchPatents

Pilot LinePrototypes

Mass ProductionProducts



Biology and µFluidics Lab: competences

In Lab & On siteValidation

Bio Protocols in µsystemsSample Preparation

Simulation Microfluidics Biology

Cytotoxicity

…for your application in

Diagnostics, Therapeutics, Environment, Homeland Security, Wellness…

System Integration

Silicium, PDMS, glass,plastic microfluidics

Soft, FlexibleMicrofluidics



We address the technological steps

Step1 : Lab Automation

Diagnostic lab

One shot

Control lab

Step 2: Lab Delocalisation

Monitoring



We can help you at different levels

InterfaceInterface

System IntegrationFirst Prototype

Proto Validation

Module



Our Fluidics Technologies…

Samplevolume

AirLiters to m3

Digitalmicrofluidics

Channelmicrofluidics

MinifluidicsContinuous flow microfluidics

‐ Fluidics technology ‐we cover the mL‐nL liquid range

Soft, flexible Fluidics

Airsampling

100 mL 1‐10 mL 100 µL 1 µL 1 nL

Nanofluidics



…Prototypes…

Molecular Biology (PCR)• Automated

• Sample : air, liquid (1‐20 mL)

• Sensitivity : 10 pathogens / 90 min

• Multiplex : up to 100 analyses

• W/V : 10 kg / 40 L

Immuno analysis• Automated

• Sample : liquid, gaz (explosive)

• Sensitivity : 30 pg to few ng (toxines)

• Multiplex : up to 10 analyses

• W/V : 2 kg / 10 L

Virus Bacteria Spores Toxines

InterfaceInterface

One shotor

Monitoring



… and their Validation with PartnersAir sampling of moldsIn a culture field (INRA)

Air sampling of sporesin a L2 lab at Robert Koch Institute (RKI)

Genomics sample preparation cartridge validationat “Hospice Civil de Lyon” lab (HCL)

Air Biomonitoring System evaluated in CSTB installation



µFluidics Toolbox for Bio AnalysisAir

Sampling

WaterSampling

BloodSampling

Pre Analytics“ How to transfer the analyte as fast, as concentrate and as “pure”

as possible on a sensor “

Sampling“ How to collect sample and

maintain its integrity “

Sensing“ How to detect, identify and/or quantify analytes “

Signal processing / Communication / User Interface“ How to extract data and deliver the answer at the right level (number, curve, color) and the right place (on site or far away) “

Immunology

Genomics

Spectroscopy(MS, Raman)



Sample prepcartridge

NO SYSTEM OFFER

for environmental

samples(volume, spore

lysis)

POTENTIAL OF INNOVATION

PerformanceUsability

Individual(< 1 kg)

Field(1‐5 kg)

Lab(10‐30 kg)

FieldMonitoring(30‐300 kg)

Size

Module

Sensing Pre Analytics Sampling

Some Challenges for µFluidics



Air Sampling

Particules 250 nm

Personal Biodosimeter• Personal system (400 g)

• No mobile parts

• Collection rate : 5‐10 LPM

Portable Air Sampler• Portable (5 kg)

• Collection rate : 50 ‐ 100 LPM

Electrostatic Air Sampling• Not time limited / continuous sampling

• Efficiency > 85% yield in a large range (10 nm to few µm)

• Viability of micro organism is conserved



Blood Droplet Sampling / StabilizationPlasma Separation• Passive system on 50 µL blood

• Plasma separation by filtration on soft materials

• Protein conservation for further extraction / analysis

membrane

Protein saver

prefilter

Blood

-50

150

350

550

750

950

1150

1350

1550

1750

11,2

5

12,1

12,9

5

13,8

14,6

5

15,5

16,3

5

17,2

18,0

5

18,9

19,7

5

20,6

21,4

5

22,3

23,1

5 24

24,8

5

25,7

26,5

5

27,4

28,2

5

29,1

29,9

5

30,8

31,6

5

32,5

33,3

5

34,2

35,0

5

35,9

36,7

5

37,6

38,4

5

39,3

40,1

5 41

41,8

5

42,7

43,5

5

44,4

45,2

5

46,1

46,9

5

47,8

picprep 23_07_07

plasma centrifugé d1/25

picprep standard J+10

picprep standard J0+5

picprep standard J0

Evolution of protein profileBetween Day 1 and Month 2



Pre analytics / Separation

Plasma separation• Plasma Quality compared to reference method (centrifugation)

• No hemolysis No protein loss

•Extraction yield ≈ 25% indepent to flow rate (50‐200 µL/min)

Hydrodynamic systems• Compatible with high volume samples (1 mL in 6 min in this example)

• No actuation : no valves, no electrodes

PlasmaOUT

CellsOUTBlood

IN



Check valve Pressurized air

Blood separation

Air

SAMPLING CELLS SEPARATION

Lab on chip

Pre analytics / Separation

Blood Analysis Protocol

Also efficient for Pathogens Concentration

• Efficient on particules > 2‐5 µm

• Spores concentration : x 2

• Yeast concentration : x 100



DNA Purification

Spores & BacteriaConcentration

Pre analytics using Silicon pillarsMicro Sample Preparation using Silicon Pillars• Microfluidic platform for bio / chemical protocols in the µL range

• A generic technology : Silicon Pillars

• Co‐integration : capture, separation, bio interaction (bacteria, proteins, DNA…)

• Function controlled by surface functionalization

PeptidesSeparation



µFluidic Cartridge for DNA profiling

DNA extractionSTR sequences

amplificationAmpliconspreparation STR analysis

Blood dropletSaliva Capillary Elec.

ABI PRISM® 3130Micro CapillaryElectrophoresis

Liu

Mini CapillaryElectrophoresis

Hopwood

Plex-ID IBIS (ESI) GeneTrace?

2001… (MALDI)

Cha RFMP adaptation dessalage

(MALDI)

Fenaille (ESI)Sample Preparation Cartridge for MS• Goal: Delocalize DNA profiling outside central labs

• Sample prep cartridge for MS detection

• MS gives access to DNA sequence (not only length)

• All pre analytics steps in less than 90 min

• Cartridge under development



µFluidics cartridge for Sepsis diagnosticsSample Preparation Cartridge for PCR• Fully integrated / automated protocol for Gene Expression

• From blood sample to RNA in 20 min

• Next generation : RT‐PCR integration

ARNm

10 to 50µL

Blood

100µL

20 min

RC

WC

m. beads1/ White blood cellextraction

2/ lysis

3/ mRNA extraction

4/ Purification

5/ Elution

Gene Expression Profile development



« All‐in‐One Cartridge » for Environment Monitoring(industry, security)

PathogensConcentration

PathogensLysis

DNA Purification

QuantitativePCR

PCRpre amplification

Sample(1‐10 mL)

OR



4 prototypes between 2012 and 2015To support technologies already on sales


PathogensLysis

QuantitativePCR

DNA Purification

PCRPre amplification

2012

D1 : Liquid/liquid

concentrator

(15 min)

Year

2013

2014

2015

Prototype

D2 : D

NA extractor

(30 min)

D3 : Sampeprep

cartridge (45 min)

D4 : «

All in

One »

Cartridge

(60‐75

min)




PathogensLysis

PathogensLysis

DNA Purification

PCRPre amplification



Thank youGuillaume Delapierre

CEA Leti / DTBS

[email protected]

+33 4 38 78 23 45

µfluidics: a toolbox for analysis automation and … › cluster › event › event › 20121004...

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