nanobiotechnology introduction - chalmers · nanobiotechnology . example: biophotonics: – science...

Chalmers University of Technology

NANOBIOTECHNOLOGY

INTRODUCTION

Thanks to Profs. O. Orwar (Chalmers) and N. Pedersen, (U. of Alberta) for slide materials


We read a lot about Nanotechnology. WHAT IS IT, REALLY?

Gary Stix, Scientific American, sept. 2001


1dm length of a finger

1cm Diameter of a finger

1 mm The thickness of a nail

100 µm Diameter of a hairstraw

1 µm Diameter of a cell nucleus

Diameter of a organelle 100 nm

Size of a large protein 10 nm

Size of an amino acid 1 nm

Length of a chemical bond 1 Å

10 µm Diameter of a cell

Nano=10-9 m

Molecules


Nanos=dwarf (greek). Nano as prefix=10-9 Nanotechnology is for a large part control and manipulation of matter on the nanometer scale, i.e. the size of individual large molecules!


Nanoscience and Nanotechnology1

• NANOSCIENCE: “..the study of phenomena and manipulation of materials at atomic, molecular and macromolecular scales, where properties differ significantly from those at a larger scale”

• NANOTECHNOLOGY: • “..the design, characterization, production and

application of structures, devices and systems by controlling shape and size at nanometer scales”

1 July 2004 a report commissioned by the Royal Society, UK (http://www.nanotec.org.uk/report/Nano%20report%202004%20fin.pdf)


Faster computers New computer architectures New electronics New optics New materials New research tools New knowledge!

Nanotechnology-Deliverables Several decades from now, we will see our current silicon-based microelectronics supplanted by a carbon-based nanoelectronics of vastly greater power and slope.

Richard E. Smalley (Nobel prize in 1996 for C60)

Important breakthroughs STM AFM Single molecule spectroscopy Micro/nanofabrication Nanomaterials e.g. carbon nanotubes Single-atom manipulation


Nanobiotechnology

Example: Biophotonics: – science of generating and harnessing light (photons) to image, detect and

manipulate biological materials.

– is used in BIOLOGY to probe for molecular mechanisms, function and structure.

– is used in MEDICINE to study tissue and blood at the macro (large-scale) and micro (very small scale) organism level to detect, diagnose and treat diseases in a way that are non-invasive to the body.

applies the tools and processes of nano/microfabrication (nanotechnological achievements) to build devices for studying biosystems (living systems!).

Convergence of Disciplines! Translational Technology Transfer from the

Physical Sciences to the Life Sciences


We want to understand biology at greater

level of detail!

Well-the first is to undertand the important features of the system we are studying!

What do we want to study?


Chemical reactions per cell and second x 1015 cells 1 million =1021 CIPS

(compare TeraFLOP supercomputer=1012 instructions/sek)

new proteins per second 1 billion 1,000 – 100,000 copies of a particular protein

~ 30,000 genes ~ 100,000 gene products > 1,000,000 post-translationally modified proteins


Common features of living systems: 1. Small scale

2. Complex geometries

3. Complex chemistry

4. Complex materials

5. Change and dynamics

6. Far away from chemical equilibrium

Complex systems-complex interactions

To gain a detailed understanding of such systems-supersensitive and small-scale interrogation and

measurement methods are required


Examples of Challenges • Molecular:

– Structure analysis of single proteins – Sequencing single DNA – Understanding transport and sorting in the Golgi apparatus

• Cellular: – Molecular scale imaging of single, living cells – Single molecule biochemistry in single, living cells

• Medical: – Finding single, abnormal cells among healthy ones in living

tissues – Developing non-invasive medical tools – Understanding the cellular biochemistry of the brain


What are the eight most important problems to be solved?


1. Spatial Resolution

Molecule Molecular Structures Organelles Cells 1 nm 10 nm 100 nm 1 µm 10 µm

Proteins Ion Channels Nanotubes Mitochondria Nucleus

Imaging: Atomic Force Microscopy

Electron Microscopy

Optical Microscopy

Spectroscopy: Optical (Fluorescence) Spectroscopies

Mass Spectrometry (SIMS)


2. Temporal Resolution ps - Bond vibrations Triggering Physics ns - Conformations µs - Binding Associations Chemistry ms - Reactions Flux s - Regulation Transport ks - Movement Regeneration Biology Ms - Development Fetal Growth Gs - Life cycle Disease


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3/4. Heterogeneity and Complexity

Many compartments, many structures, many surfaces As many as 85% of biochemical interactions occur at or in a membrane


5/6. Specificity and Discrimination We need to be able to measure pair-wise interactions:, e.g.

sequentially – exclusively and whether transient or long term stable

cis and trans interactions among the 22-member γ-protocadherin family ("calcium-dependent adhesion”), which have been shown to be critical for the control of synaptogenesis and neuronal survival.

We need to be able to discriminate between monomers, oligomers and polymers: Single molecule studies requires research platforms for single molecule investigation


7. Sensitivity-Detection Limits Typical

Concentrations in a cell or Densities on a cell surface

N = 100,000 molecules

V = 10x10x10 µm3

= 1000 fL = 1 pL

C = 100 molecules per µm3

C ~ 100 nM

N = 100,000 molecules

A = 10x10x6 µm2

= 600 µm2

C ~ 100 molecules per µm2

Required: single molecule detection sensitivity for sub-micron resolution

Fluorescence can provide 100 counts per molecule per millisecond


8.Data processing and storage

1 CD per image 1 hard drive per experiment

m/z m/z

One image can contain 512x512x2000 = 512Mb intensity measurements

Example: Particle beam interaction using ToF-SIMS. Incident particles bombard the surface liberating single ions (+/-) and molecular compounds


To approach the problems we need to think about… How to reach nanoscale biological objects?


Integrative Nanoscience ● Nanoscale materials and objects must be integrated into microscale systems that interact with the outside world.

● Suitable observation and manipulation tools are needed


Reality check!

2011


2012


3D vs. 2D Fluidic Systems

orientational disorder orientational order

positional disorder positional disorder

high pressures necessary no bulk pressure or bulk solvent flow

lower size limit >> molecule size easily scaleable

Squires and Quake (2005) Rev. Mod. Phys. 77, p. 977ff.

Kam and Boxer (2003) AdFunct. Mater. 16, p. 306ff.

2D Nanofluidic Devices


2D Nanofluidic Devices Ternary mixer Spreading on nanolanes

and SAM release

Microfabrication: UV lithography

Nanofabrication: direct e-beam writing

Lift-off techniques

Lipid vesicle manipulation


H2O

hydrophobic support

Lipid bilayer in multilamellar vesicle

spreading

hydrophobic tail hydrophilic headgroup

Phosphatidylethanolamine

hydrophobic supports: SU-8, EPON 1002F, Teflon AF

Lipid Adsorption on Hydrophobic Surfaces Lipid Monolayer Spreading

www.mardre.com An electron microscopy micrograph of a MLV. Scale Bar: 1 µm

http://www.mardre.com/�


2D fluidic system

2D Reaction Platform

Surface profile

Mixing junction

Sites for vesicle deposition

Detection

Small scale reaction systems with few molecules

2D reaction vessel

Slide Number 1Slide Number 2Slide Number 3Slide Number 4Nanoscience and Nanotechnology1Slide Number 6NanobiotechnologySlide Number 8What do we want to study?��Slide Number 10Slide Number 11Examples of ChallengesSlide Number 131. Spatial Resolution2. Temporal Resolution3/4. Heterogeneity and Complexity5/6. Specificity and Discrimination7. Sensitivity-Detection Limits8.Data processing and storage Slide Number 20Integrative NanoscienceReality check!Slide Number 233D vs. 2D Fluidic Systems2D Nanofluidic DevicesLipid Adsorption on Hydrophobic Surfaces�Lipid Monolayer Spreading2D Reaction Platform

nanobiotechnology introduction - chalmers · nanobiotechnology . example: biophotonics: – science...

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