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Bioinspired of Micro-fluidic systems Reach Symposium-2008. Shantanu Bhattacharya Assistant Professor Department of Mechanical Engineering Indian Institute of Technology Kanpur bhattacs@iitk.ac.in Tel: 0512-259-6056. The field of Bio-mimetics.

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bioinspired of micro fluidic systems reach symposium 2008

Bioinspired of Micro-fluidic systems Reach Symposium-2008

Shantanu Bhattacharya

Assistant Professor

Department of Mechanical Engineering

Indian Institute of Technology Kanpur

bhattacs@iitk.ac.in

Tel: 0512-259-6056

the field of bio mimetics
The field of Bio-mimetics
  • Quite often Nature works at Maximum Achievement at minimum effort level.
  • The field of Bio-mimetics is the abstraction of a good design from nature.
  • The field of Bio-mimetics started from Giovanni Borelli’s seminal De Motu Animalum of 1680.
blood capillaries and micro fluidics
Blood Capillaries and Micro-fluidics
  • The capillaries are the smallest blood vessels that distribute oxygenated blood.
  • Inspired by the micro-circulation inside the capillaries and its uses one can think of Micro-fluidics
slide4

Micro-fluidics

  • Properties of Micro flows
  • Surface effects become prominent with high surface area to volume ratio.
  • Low thermal mass and high heat transfer.
  • Low value of Reynolds number and thus laminar flows which only result in diffusional mixing.
  • Re is usually less than 100 and often less than 0.1 in micro-devices
materials in micro fluidic devices
Materials in Micro-fluidic devices
  • Silicon and microelectronic materials
  • Glass, Quartz

Alternate Biochip materials

  • Polymers

– Poly (dimethylsiloxane) (PDMS)

– Poly (methyl methacrylate) (PMMA)

– Teflon, etc.

• Biological Entities

– Cells, Proteins, DNA

– Frontier of BioMEMS !

slide6

Microfluidic device fabrication in Silicon

  • NEMS/ MEMS silicon fabrication
  • Conventional and new semiconductor manufacturing techniques are used.
  • Etching, Deposition, Photolithography, Oxidation, Epitaxy etc.
  • Deep RIE, Thick plating etc.
  • Bulk and surface micromachining.
micro channel arrays using controlled etching
Micro-channel Arrays using Controlled Etching

Real image of micro-channels

after swelling in solvent

Ref: Sharma et. al., Science, 2007

1- Dimensional

2- Dimensional

3- Dimensional

Cross-sectional View

slide9

Micro-fluidics and Microsystems

Relationship with the Biological world

  • Systems made up of very small components.(micron to nanometer scale)
  • Relatively high applicability to the field of life science, biotechnology and medicine.
  • That’s why they scale with some of the biological entities.
  • Focus of micro-system research is shifting to micro fluidic systems.

Bottom up

Ref :Stephen D. Centuria, Microsystem Design, Kluwer Academic Publishers, Boston / Dordrecht / London

Lectures, from NanoHUB, Purdue University, West Lafayette, Indiana

content of presentation
Content of presentation
  • Mimicking Biological architectures for certain engineering end goals.
  • Mimicking Biological principles for certain engineering end goals.
  • Micro-fluidics and Bio-sensing
slide11

Micro-fluidic Sample Preparator

  • A micro-separation device is realized
  • Whole blood enters the device through a 70 microns channel.
  • Margination happens and leukocyte distribution is affected .

Bitensky et.al., 2005, Anal. Chem. 77, 933-937

microfluidic tectonics a comprehensive construction platform for microfluidic systems
Microfluidic tectonics: A comprehensive construction platform for microfluidic systems
  • There are a lot of passive valves in our veins , allowing the fluid flow in only one direction.
  • Hydrogel is used to realize a valve which swells and de-swells in different pH’s

Beebe et. al., 2000, PNAS, Vol. 97, pp. 13488-13493.

characteristics of bacterial pumps in microfluidic systems
Characteristics of bacterial pumps in microfluidic systems
  • The growth of flagellum in flagellated bacteria like E. Coli or Serratia Marcescens is a function of glucose conc.
  • Flagellated bacteria are used in microchannels to paddle fluids at various flow rates.

Fig. 1

Fig. 2

Fig. 3

Kim et.al., NSTI-Nanotech 2005, Vol.1

biochips driven by bioinspired microfluidics
Biochips driven by Bioinspired Microfluidics

Bhattacharya et. al., JMEMS, 2007.

Chang et. al., Biomedical Microdevices, 2003.

Bhattacharya et. al., Lab chip, 2007, under review.

lab on chip for daignostics of infectious bovine rhinotracheitis
Lab on chip for daignostics of Infectious Bovine Rhinotracheitis
  • Annual losses due to the bovine viral disease IBR to the Beef industry stands at US$ 10-40 million per million animals (Bennett & Done, 1992,Harkness, 1997, Houe et al., 2003b). or $560million per annum. http://www.livestock.novartis.com/pdf/Arsenal_BVD_KnowlEdge.pdf
  • Originally recognized as a respiratory disease in swine herds in 1991.
  • Mechanism of transmission are mainly confinement particularly in feedlots. The disease is rapidly spread to new arrivals for already infected species.
  • Field diagnosis is extremely important.
  • Detection is carried out using PCR based assay in laboratories which is time consuming.

Ref: Infectious Porcine Diseases, L.R. Sprott and S. Wiske, Agricultural communications, 2002

dna translation in agarose electrophoresis
DNA translation in Agarose (Electrophoresis)

I

1

+ve

-ve

II

Sequential fluorescent images of DNA migration behavior in mediums: (a) Nanospehere (b) Agarose and (c) Control Buffer solution without nanosphere 1

+ve

-ve

[1] Nanospheres for DNA separation chips

Mari Tabuchi1, 5, 6, Masanori Ueda1, 5, Noritada Kaji1, 5, Yuichi Yamasaki2, 5, Yukio Nagasaki3, 5, Kenichi Yoshikawa4, 5, Kazunori Kataoka2, 5 & Yoshinobu Baba1, 5, 6, 7 , NATURE

III

equipment in a pcr laboratory
Equipment in a PCR laboratory

DNA Extraction from tissue samples

Imaging of Fluorescence

Glove box for preparing the PCR Mix

Gel electrophoresis of DNA

PCR thermal Cycler

slide23

Lab on Chip Design for the Analyzer

Non fluorescing reference channel for background subtraction

Syringe for injecting PCR mix and sample

Electrodes for Gel electrophoresis

Micro channel filled with agarose gel

Compressed air bottle

LED

Plan View

Front Elevation View

Spectrometer

Reference Solid Core Waveguide for background subtraction

Solid Core Waveguides placed along target DNA regions

Lab-view Operated Solenoid valve

Heaters for PCR

DAQ system hooked to spectrometer will provide the spatial data for the differential intensities

Optical Fibers from Assay

Computer with DAQ card

Spectrometer

slide24

Peristaltic Micro-pumps for fluid transport

  • Peristalsis is the motion of fluid in channels through a traveling contractile.
  • This effect has been successfully utilized for the control of fluid motion.
  • Pumping rates in the range of 10~12 microliters at pumping frequency of 10 Hz. has been attained.
  • The pumps are 3 layered devices fabricated using Glass and PDMS and are energized by an offchip compressed nitrogen supply regulated thru labview.

Pneumatic Chambers

Inflow

Fluid Channels

Outflow

Pumping Cycle

Pumps in action

Picture of the pumps

working pcr chip for ibr isolates
Working PCR Chip for IBR isolates

Amplified Extract from chip

Amplification performed on .07 pg/ μl sample conc.

Amplified Sample from Conventional M/c

Ref: “Optimization of design and fabrication process for realization of a PDMS-Silicon DNA amplification chip”, by Shantanu Bhattacharya, Venumadhav Korampally, Yuanfang Gao, Maslina Othman, Sheila A. Grant, Steven B. Klieboeker, Keshab Gangopadhyay, Shubhra Gangopadhyay”, Journal of Microelectromechanical systems,Vol.99, pp.1-10, 2007.

capillary electrophoresis sample and capillary loading
Capillary Electrophoresis: Sample and Capillary Loading

2 Basic Capillary Designs

A

B

Sample loading sequence in Gel filled channels

capillary electrophoretic chip reduces detection time by a factor of 40
Capillary Electrophoretic Chip Reduces Detection Time by a Factor of 40

1.5% agarose solution in microchannels

300 V for 50 secs

300 V for 25 secs

DNA ladder Trial: 100-1000 bp movement in an Agarose capillary.

Mobility (μ) = 9.101E-4 cm2/ Vsec .

Velocity of the stain=.078 cm/sec

Electric field = 85.7 V/cm

Conventional Electrophoresis Time= 35mins

Requirement : Low voltage capillary electrophoresis system

Ref: Bhattacharya, S., Gangopadhyay K., Gangopadhyay, S., Sharp, P.R., “A low voltage capillary electrophoresis system using platinum doped agarose gels”, (Manuscript to be submitted to Biosensors and bioelectronics).

low voltage electrophoresis by applying nanotechnology
Low Voltage Electrophoresis by Applying Nanotechnology

Platinum nano-particles made in situ

Potassium Chloro-Platinate is reduced by sodium boro-hydride after coating with a monolayer of Mercapto-Succinic acid in a Schlenk line in inert Argon environment. (2 conc. of solution used are 11.6mM and 23.2mM)

Average particle size= 13.16nm,

+ 3.93nm

sem tem images of the doped gels
SEM/ TEM images of the doped gels

TEM image of Platinum doped agarose

Back scattered image (FESEM)

Array image of platinum particles embeded in agarose

Sizes:

2.5 microns

500 nm

500 nm

Ack.: Lou Ross, Randy Tindel and Cheryl Jensen., EMC core

EDS spectra of the Platinized gels

enhanced dna mobility
Enhanced DNA mobility

Mobility Enhancement 2 times at 16V/cm

Calculations done using the one dimensional mobility model

µ = v/ E

where , µ = mobility of the stain, v= Velocity (cm/ sec.), E= Electric Field (V/cm)

dielectric constant enhancement due to nano platinum

Cdi

Rser

Rs

Zw

Zw

Dielectric Constant Enhancement due to nano-platinum

Electrode spacing

= 23microns

Electrode width= 17microns

Mobility = ε ε0 ζ / η [1]

εhas approx. 2 times enhancement

[1] Rieger T.H., “Electrochemistry”; Prentice Hall, inc., New Jersey, 1987

Ref: Bhattacharya, S., Chanda, N., Grant S.A., Gangopadhyay K., Gangopadhyay, S., Sharp, P.R., “High conductivity agarose nano-platinum composites”, (Manuscript under review in Analytical Chemistry).

summary and conclusions
Summary and conclusions
  • Bio-inspired Micro-fluidic technology is widely applied for biomimetics and biosensing.
  • Lab-on-Chip is a direct spinoff of this technology and is used for providing point of care diagnostics.
  • There is huge market potential for these technologies for the numerous applications.
acknowledgements
ACKNOWLEDGEMENTS

Collaborators and Advisors:

  • Dr. Sheila Grant, Dr. Shubhra Gangopadhyay , Dr. Keshab Gangopadhyay, Dr. Steve Klieboeker, Dr. Lela Riley, Dr. Xudong Fan (University of Missouri, Columbia).
  • Dr. Rashid Bashir, Dr. Arun Bhunia, Dr. Michael Ladisch. (Purdue University, Indiana).
  • Dr. P. Panigrahi, Dr. Bikram Basu, Dr. Bishakh Bhattacharya (IIT- Kanpur).

Funding Agencies:

  • NSF (Curriculum Research Curriculum Development).
  • NPB (National Pork Board).
  • NIH (Mutant Mouse).
  • USDA (Center for food safety engineering).
  • Initiation Grant (IIT-Kanpur, DORD)