Lecture 2 Introduction to BioMEMS & Bionanotechnology

Biomedical Innovations by BioPOETS*

*BioPOETS: Biologically-inspired Photonics-OptofluidicsElectronics Technology & Science

Luke P. Lee
Department of Bioengineering

Physiologically Relevant Fluidic ICs

for Quantitative Cell Biology and Quantitative Medicine on Chip

The BioPOETS UC Berkeley

The BioPOETS UC Berkeley

Cellular BioASICs*
*Biological Application Specific Integrated Circuits
BioASIC #1 Integrated Microfluidic Patch-clamp Array Chip BioASIC #2 Single Cell Electroporation Chip BioASIC #3 High-density Single Cell Analysis Chip BioASIC #4 Dynamic Cell Culture Chip for Systems Biology BioASIC #5 Cell-cell Communication Chip BioASIC #6 Cell Lysing Devices for Sample Preparation BioASIC #7 Biomimetic Cell Sorting Microfluidic Devices BioASIC #8 Micro PALM for Cell Manipulations BioASIC #9 Integrated Cell Culture & Lysing & Harvesting BioASIC #10 Biomimetic Artificial Livers on a Chip BioASIC #11 Biofluidic Self-assembly of Spheroids on a Chip http://biopoets.berkeley.edu

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Basics of BioASICs: Fluidic Resistance

z x y h w Pressure Driven Navier-Stokes L

Time, translation invarience at low Re

Cross Section 2x5 m 50x50 m

R/L (Pa s/m3) 4.0*1020 7.5*1014

R1/R2 = 5.4*105

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Sample/Fluidic Interface

Soft State BioASICs

Electrical Interface Fluidic Interface

Cell Culture BASIC

Cellular BASIC

nSERS-based Molecular BASIC

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BioASIC #1

Integrated Microfluidic Patch-clamp Array Chip

(IMPAC)
Sigworth, Nature 423, 21-22 (2003)

Mammalian Electrophysiology on Microfluidic BioASICs* Platform

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BioASICs: Biological Application Specific Integrated Circuits

C. Ionescu-Zanetti, R. M. Shaw, J. Seo, Y. Jan, L. Y. Jan, and L. P. Lee (PNAS, 2005)

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Biologic Microfluidic Pulse Generator Fast Solution Exchange

Pulsed Reagent Flow

-P
Constant Buffer Flow

Understanding kinetics of ligand-receptor interactions

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Simultaneous Electrophysiological and Optical Measurements

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Ion Channel Gating (Kv2.1)

Voltage gating Ligand gating

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BioASIC #3

High-density Single Cell Analysis Chip

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High density Single Cell Array via

Hydrodynamic Single Cell Tweezers

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Quantitative Single Cell Biology

The BioPOETS UC Berkeley

The BioPOETS UC Berkeley

Dynamic Single-Cell Microfluidics for Long-term Cytotoxic Drug Assay

Single Cell Culture Chamber Media Input

Drug Input

log-gradient generator

Media : Drug = 9:1

Waste Output

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BioASIC #4

Physiologically Relevant Dynamic Cell Culture System:

Cultural Revolutions

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Biomimetic Physiological Microenvironment

c i D
Blood Cell Flow Loading
Cell Growth and Interstitial Space

Tissue Size (D) Circulatory Flow (c) Interstitial Flow (i) Extracellular Matrix 100-300 m 700 m/s 0.1 m/s Complex

Microfluidic 50-1000 m 80-4,000 m/s 0.08-4 m/s Surface Coating

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Cell Culture Biotechnology

Volume Cell Density (v/v) Medium Turnover Throughput

Microtiter Plate 50 l 5% 3 days 1-384

CSTR Microfluidic Bioreactor Bioreactor 2L 3 nl <10% 0.5-3 days 1 40-80% 2-120 sec 64-1024

Nanoliter Scale Microbioreactor Array for Quantitative Cell Biology

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Lee et al., Biotechnology & Bioengineering, 94 (1), 5-14 (2006)

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Design for Uniform Fluid Velocity Profile

High Aspect Ratio between the Perfusion Channels & Cell Culture Chamber

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Uniform Mass Transfer

a

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CHARM* Cell Culture Well

Addressable Channels

*CHARM: C-shaped High Aspect Ratio Microfluidic

Ro a Concentration Gradient hc hr Ro Cell s R Ro

r

Ro b c d
Fig. 1. (a) Brightfield image of a 64 unit microfluidic cell analysis array. Each well has a culture area 200 m in diameter. A microfluidic concentration gradient generator is depicted on the left, providing a separate reagent concentration to each row. The 8 columns are individually addressable. (b) SEM image of a single CHARM culture well with channel height hc = 50 m and ring height hr = 2 m. (c) Finite element simulation of flow rate through the single culture unit. (d) Equivalent circuit model of fluidic resistances of the single unit microfluidic design.

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Cell Growth

(HeLa Cell, 30 min/frame, 38 hours)

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Quantitative Data

Microfluidic
24 Well Plate

Cell Attachment Kinetics

Cell Growth Rate

Cell Types Cultured

The BioPOETS UC Berkeley

HeLa

HeLa tumor

NIH3T3 Fibroblast

Primary BAEC

HepG2 Hepatocyte

SY5Y Neuroblasts

BioASIC #9

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Integrated Microfluidic Cell Culture & Lysis on a Chip

J. Tanner Nevill, Ryan Cooper, Megan Dueck, and Luke P. Lee (LOC 2007)

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Cell Culture/lysis Platform

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OH as Lytic Agent

RBCs, fast lysis event t = 60 ms

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BioASIC #10

Physiologicallyinspired Artificial Liver Sinusoids

Lee et al., Biotechnology and Bioengineering 97, 1340 (2007)

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Liver Micro-architecture

Sinusoid space transports blood to hepatocytes Lined with fenestrated endothelial barrier Hepatocytes form extensive cell-cell contact

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Artificial Liver Sinusoid

Precision Control of Hepatocyte Loading

Microfluidic endothelial barrier High density hepatocyte culture Continuous flow mass transport

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High density = happy cells

Effect of Hepatocyte Density

Low density = dead cells

Microfluidic culture without ECM coating Spheroid effect previously documented

Collagen coating required for dish based hepatocyte culture High density loading can rescue viability in absence of ECM coating

Multiplexed Primary Cell Culture with Packing Density & Flow Controls for Stem Cell & Systems Biology

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The BioPOETS UC Berkeley

Optical MEMS
for Lab-on-a-Chip

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Microlens Scanner

S. Kwon & L. P. Lee, IEEE MEMS (2001)

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MEMS Confocal Microscope

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Lab-on-a-chip with CIAs

In-vivo SMM & SMD Imaging Microsystem

HTS Genomics & Proteomics Nano- & Micro-factory Lab-on-a-chip

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CIAs for Cellular Imaging

-------- 300 m

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Images from CIA

1 m resolution

High-content Integrated Quantitative Molecular Diagnostics: High IQ-MD

In-vivo IR Spectroscopy In-vivo detection window Confocal microscopy Confocal nSERS

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Cell trapping

CIAs

Cellular Analysis Cell lysing nSERS

Lab Automation: Sample Prep, SMM, & SMD Microfluidic Pumps Microfluidic interface

Cell sorting by adhesion protein

1mm

Nanogap Junction

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Innovative Personalized Medicine

Disposable Diagnostic Biochip

Mobile Healthcare iMDs*

Information Technology Biotechnology Nanotechnology *iMDs: Innovative Medical Diagnostic Systems

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Summary
Biologically-inspired photonics and optical systems are being developed for innovative healthcare systems. Cellular BioASICs are being developed for quantitative biology & medicine. Quantum nanoplasmonic molecular probes, molecular ruler, ONCOS (gene regulator & protein expression controller) are developed for molecular/cellular imaging, and quantitative in vivo biology. Using BioPOETIC 3D packaging, high-content Integrated Quantitative Molecular Diagnostic (iQMD) system can be created for future preventive, personalized medicine, and integrated health & environmental monitoring systems.