AFM Imaging in Liquids Kada 050708 - Agilent

Agilent in NanotechnologyAgilent in NanotechnologyAFM Imaging in LiquidAFM Imaging in LiquidAFM Imaging in LiquidAFM Imaging in Liquid

Gerald Kada, PhDNano-Measurements

DivisionLinz AustriaLinz, Austria

Group/Presentation TitleAgilent Restricted

Month ##, 200XPage 1

Design Consideration of the AFM ScannerDesign Consideration of the AFM Scannerfor Imaging in Liquid Environmentfor Imaging in Liquid Environment

• Top down scanning:

-> no electronics under the sample plate

• Pendulum design and separated piezo• Pendulum design and separated piezo plates for unperturbed scanning in xyz, low mechanical noise, fast scanning

• Atomic and Molecular Resolution

• Open liquid cell / closed cell

• Magnetic AC (MAC Mode) forgentle imaging in liquid



Tip Scanning AFMTip Scanning AFM

Closed-loop XY scanning stageUntraditional sample sizeU t ad t o a sa p e s e

Heating /CoolingFluid imaging



Heating /CoolingFluid imaging

Soft Imaging in SolutionSoft Imaging in Solution

Acoustic AC mode Traditional way:

(“tapping“)

A piezoelectric transducer shakestransducer shakes the cantilever holder

MAC ModeAgilent way:

Cantilever is coated with magnetic filmA solenoid applies an AC field to bend cantilever



field to bend cantilever

Performance Bench Mark: “Tough” conditionsPerformance Bench Mark: “Tough” conditions

Example 1: Cu UPD (underpotential deposition)Challenge: Provide atomic resolution of electro-chemically controlled deposition of a copper monolayer

Example 2: Lambda DNA in liquid (Mg2+ buffer)Challenge: See the movement of DNA on a second scale in buffer environment

500nm xy sizeon gold in sulfuric acid 500nm xy size

Cumonolayer

Auatoms Image conditions:

MAC mode (controlled low force)



48 lines/sec, 4 secs for full image

SaltSalt--induced ‘melting’ of induced ‘melting’ of MMTV promoter chromatinMMTV promoter chromatin

Plasmid DNA in buffer solution450 nm scan sizeKienberger et al, Biomaterials 2007

pp

(DNA(DNA--protein complex)protein complex)

1 4M

0M

1.4M

NaCl

0M0.8 x 1 µm scans

Wang, Lindsay et al, Biophys J (2002)



g, y , p y ( )

PVP PVP -- Polyelectrolyte BrushesPolyelectrolyte Brushes

M l l Ch t i ti• Molecule Characterization• Film Formation

- Polymer concentration- ionic strength- solvent

1 μm1 μm μμ

t = 81 st = 81 s

in aqueous solution • stiff conformation

390 nm

Γ = 45 %after adding NaCl



aqueous so ut o st co o at o• width: 8 nm, heigth: 2.8 nmΓ = 20 % Γ = 45 %g

Live Human RhinovirusLive Human Rhinovirus (MAC Mode in liquid)(MAC Mode in liquid)

RNA release from virus by decreasing pH



Kienberger, Hinterdorfer et al, J Virology (2004)& Structure (2005)

… on Top of a Living Cell in Dynamic Force Mode… on Top of a Living Cell in Dynamic Force Mode

Live endothelial cells MAC Mode in PBS MAC Mode is perfectLive endothelial cells, MAC Mode in PBS buffer solution(Left) 70 µm scan, 5 µm height

(courtesy of K. van Vliet, MIT, USA)(Right) 13x20 µm scan, 150 nm height

for imaging DNA,Proteinsor Cells under physiological conditions



(courtesy of Ch. Riethmüller, Uni Münster, Germany)

MultiMulti--frequency (Higher Harmonics) imagingfrequency (Higher Harmonics) imaging

Imaging at different harmonics is suggested for clarification of contribution of various nonlinear (mechanical and adhesive) tip-sample interactions to the image contrast and the quantitative estimate of these properties



the image contrast and the quantitative estimate of these properties.

Higher Harmonics imagingHigher Harmonics imaging (MAC mode in liquid)(MAC mode in liquid)Bacterial SBacterial S--LayerLayer

Topography (a-left) and2nd Harmonic image (a-right)

(b) Averaged image of 55 units

<1 nm resolution on protein crystal in liquid (MAC)



Preiner et al, PRL 20071 nm resolution on protein crystal in liquid (MAC)

3 pm amplitude sensitivity in 2nd harmonics

Finally…Thanks for your attention !y

AcknowledgementsAcknowledgements

- University of Linz, Biophysics InstituteProf. Peter HinterdorferFerry Kienberger (now Agilent Labs)Johannes Preiner Agilent Technologies Inc.

Nanotechnology Measurements Division- Czech Academy of Sciences, Nove Hrady

Prof. David KaftanAlexander Dulebo

gyTianwei JingJohn AlexanderJoe CrabtreeSergei Magonov

Contact:[email protected]/find/AFM

- Arizona State University, Biodesign InstituteProf. Stuart LindsayHongda Wang



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AFM Imaging in Liquids Kada 050708 - Agilent