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Autonomous cell-based biosensor microsystem

A week of autonomy (?)

Communication unit (to GSM device)

Versatile for various cell models and secondary sensors

Cell-based biosensors provide a biologically relevant

response to toxic compounds and mixtures

CellsCells optical

sensors

nutrient

supply

signal

processing

unit

comm.

unitelectrical

sensors

sampling

data com

microsystem platformbioreactor

power supply

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78" 9 +"&.' /- 4/5"6"-'" :2,;"1+

EPFL-LMIS Microsystems Laboratory Ph. Renaud

Microfluidics, cell chips, bio-impedance

UNIL-DMF Department of Fundamental Biology J. van der Meer

Cell biology, gene reporters, bacterial sensors

HESSO-ISI Industrial Systems Institute M. Geiser

Microsystems, electronics, optical sensors

ETHZ-MAT Biologically Oriented Materials V. Vogel

Biomaterial, cell biology

CSEM Nanobiotechnology group M. Liley

Surface biochemistry, biomaterials

EPFL-LEPA Electrochemistry and Analytics Lab. H. Girault

Electrochemical sensing, analytical chemistry

EPFL/NE-IMT Sensors and Actuators Laboratory N. de Rooij

Microfluidics, microsensors / P. van der Val

UNIL- IST Institute of Occupational Health M. Riedicker

Health effect of pollution

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SYSTEM INTEGRATIONSYSTEM INTEGRATION

--Systems specificationsSystems specifications

--Electronic platform + demoElectronic platform + demo

--Bioreactor integrationBioreactor integration

--Validation ofValidation ofbioreactor controlbioreactor control

--DemonstrationDemonstration

MICRO-BIOREACTORMICRO-BIOREACTOR

--Bacterial microbioreactorBacterial microbioreactor

--Biomaterials andBiomaterials and3D3Dcultureculture

--Microfluidic platformMicrofluidic platform

--Nutrient and sample supplyNutrient and sample supply

--Integration ofIntegration ofmonitoring sensorsmonitoring sensors

SECONDARY SENSORSSECONDARY SENSORS

--Electrochemical microsensorsElectrochemical microsensors

--Fluorescence/bioluminFluorescence/biolumin. sensors. sensors

--Impedance sensingImpedance sensing

--Sensors validationSensors validation

CELL MODELSCELL MODELS

--SpecificationSpecificationof cell modelsof cell models

--CultureCultureprotocolsprotocols

--AssessmentAssessmentof cell modelsof cell models

--Culture inCulture in microbioreactorsmicrobioreactors

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!"## .,("#'

"( %&1+"2/& ?@$1,#/A

B#*,2"'1"-1" ,2 "#"1+2,18"./1 ("+"1=,-

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LMIS

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LMIS

LEPA

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O"+"1=,- '3'+". /' %&'"( ,- +8" /-+"2- 2"0*#&=,- ."18&-/'. ,D %&1+"2/&

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W

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U&1+"2/ 1"##' /- ./12,T*/(/1'

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@:/+8"#/ 1"##' &-( 7@@F

RTGill-W1 cell line (fish)

Cultured between 18-21C

Grow under atmospheric CO2 conditions

Cell line did NOT form tight junctions

Caco-2 cells (intestestine cell line)

Known high TEER values Looking for primary fish cell lines that form tight junctions

Measure Trans Epithelial

Electrical Resistance (TEER)of epithelial cells on ultra-thin

porous silicon nitride

membranes.

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@#"1+2,18"./1 '"1,-(&23 '"-',2'

Deposition Points

PBS Inlet

SampleInlet

Outlet

I/12,T*/(/1 &.:"2,."+2/1 "-S3.&=1 0#*1,'" R #&1+&+" '"-',2

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@#"1+2,18"./1 '"1,-(&23 '"-',2'

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LEPA

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J":

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J":&+,13+"' C /.:"(&-1" .,-/+,2/-0

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Cell growth

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ium

Day1

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Day3

Day4

Day5

Day6

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50

100

150

200

250

Relativeimpedance/%

0.1mM SDS toxicity

Day7

after5

min

after15m

in

after1

.5h

after2

.5h

0

50

100

150

200

250

Relativeimpedance/%

LMIS

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!"#$%& () *+(,%$-%)-% ,%$()#)-% %)%,./ 0,#)$1%,2

D: Alexa 488

A: Alexa 546

Baneyx, Baugh,Vogel, PNAS 98 (2001)14464 & PNAS 99 (2002)5139.

Smith, Gourdon, Kubow, Little, Vogel, PLOS Biology5 (2007)e268

Biochemical labelingof plasma Fibronectin,

added to cell medium,

harvestedby cells,

incorporatedinto extra-cellular matrix

donors random on lys, acceptors on cys

S SA S S

N C

NC

AD D

D DDD

DA A

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Smith, Gourdon, Kubow, Little, Vogel, PLOS Biology, 5 (2007) e268

Little, Smith, Ebneter, Vogel, Matrix Biol. 27 (2008) 451

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63'+". /-+"02&=,-

CellsCells optical

sensors

nutrient

supply

signal

processing

unit

comm.

unitelectrical

sensors

sampling

data com

microsystem platformbioreactor

power supply

7/26/2019 Live Sense 2010

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63'+". /-+"02&=,-C %/,2"&1+,2 R '"-',2'

7/26/2019 Live Sense 2010

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W*+,-,.,*' '"-',2 :#&[,2.

F"D2&1+,."+23 '"-',2 ?'*0&2 /- G/-" 02&:"'A

U#*"+,,+8 1,. +,

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SYSTEM INTEGRATIONSYSTEM INTEGRATION

--Systems specificationsSystems specifications

--Electronic platform + demoElectronic platform + demo

--Bioreactor integrationBioreactor integration

--Validation ofValidation ofbioreactor controlbioreactor control

--DemonstrationDemonstration

MICRO-BIOREACTORMICRO-BIOREACTOR

--Bacterial microbioreactorBacterial microbioreactor

--Biomaterials andBiomaterials and3D3Dcultureculture

--Microfluidic platformMicrofluidic platform

--Nutrient and sample supplyNutrient and sample supply

--Integration ofIntegration ofmonitoring sensorsmonitoring sensors

SECONDARY SENSORSSECONDARY SENSORS

--Electrochemical microsensorsElectrochemical microsensors--Fluorescence/bioluminFluorescence/biolumin. sensors. sensors

--Impedance sensingImpedance sensing

--Sensors validationSensors validation

CELL MODELSCELL MODELS

--SpecificationSpecificationof cell modelsof cell models

--CultureCultureprotocolsprotocols

--AssessmentAssessmentof cell modelsof cell models

--Culture inCulture in microbioreactorsmicrobioreactors

^"Q+ '+":' _

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4/5"6"-'" /-(*'+23 '*::,2+

Lonza Cell culture products access to cells and culture kits

Ayanda Biosystems Bioelectric platform platform, microelectrodes

Debiotech Drug delivery devices micro-pump samples

Novozymes Protein engineering, HTS access to cell based assays

Neroxis/Veolia Chemical microsensors sensor samples

Diagnoswiss Electrochemical immunosensing expertise, sensor samples

Biocartis Multiplex diagnostics protein microcarriers

Trade-4 Telemedecine technical support

+ European centre for validation of alternative methods expertise, standards