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Imaging Spr in the Ellipsometric Mode for Biomolecular Interaction Studies I

Introduction 

 

nanofilm_ep4

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Ellipsometric contrast Oil | DMPE | water

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 Frontview Nanofilm_ep4

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 Thickness map Air | OTS | SiO₂ | Si

 

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Delta map Air | Graphene | SiO₂ | Si

 

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Thickness map Air|PEDOT|ITO|PET-foil

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Psi-map Air|Photoactive layer|PET

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Perspective Nanofilm_ep4

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Delta map Air|Graphene|SiO2|Si

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Thickness map Air|PCBM(spin-coated)|gold

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Thickness map Air|PEG-SH|gold

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Knife edge illumination Air|glass

 

Measurement procedures : 

Imaging ellipsometry Delta and Psi maps

 

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Recording a Delta-map 

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Imaging ellipsometry

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maps measurement procedures: patented Regions of Interest (ROIs) concept 

 

Regions of interest

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Why nulling ellipsometry?

 

Nulling Ellipsometry advantage 1:

• highest sensitivity for ultra-thin films

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Nulling Ellipsometry advantage 2:

• highest accuracy

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Nulling Ellipsometry advantage 3:

• good for imaging – enhanced ellipsometric contrast

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applications of imaging ellipsometry 

 

Typical application of Imaging ellipsometry

 

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Typical application

antigen/antibody interaction at the surface of optislides 
Optislides 

The nanofilm_optislides are especially designed glass slides with optimal optical properties for measurements of bio-relevant layers - e.g. proteins, DNA, lipids, etc.

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Kinetic/SPR-cell with optislides 

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Association at various analyte concentrations

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Association at various analyte concentrations

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Array generato 

- software wizards for easy ‘region of interest’ pattern

- generation on symmetric rectangular microarrays

- definition of spot and reference regions

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Lipopeptides derived from HIV and SIV mimicking the prehairpin intermediate of gp41 on solid supported lipid bilayers 

<Schuy S, Schäfer E, Yoder NC, Kumar K, Vogel R, Janshoff A (2009) Lipopeptides derived from HIV and SIV mimicking the prehairpin intermediate of gp41 on solid supported lipid bilayers. Journal of Structural Biology 168, 125–136>

Time course of the D values obtained from ellipsometric measurements during the in situ coupling reaction of simian immunodeficiency Virus (S-N36) and human immunodeficiency virus (H-N369) to a lipid bilayer composed of 90% DOPC/10% MCC-DOPE (DOPC*) and subsequent adsorption of potential antagonists of the trimer-of-hairpin conformation C34 and as a function of time

 

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simian immunodeficiency Virus (S-N36)

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human immunodeficiency virus

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Supported Lipid Bilayers at Skeletonized Surfaces for the Study of Transmembrane Proteins 

<Fabre RM, Okeyo GO, Talham DR (2012,) Supported Lipid Bilayers at Skeletonized Surfaces for the Study of Transmembrane Proteins. Langmuir 28, 2835-2841>

 

SPREE analysis of BK ion channel incorporation into lipid membranes

 

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Free vesicles were then removed with buffer rinsing

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BK ion channel incorporation into lipid membranes supported on zirconium phosphonate modified surfaces

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The membrane was again rinsed with buffer. The buffer used was trizmahydrochloride and sodium chloride at pH 7.4..

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Differential Protein Assembly on Micropatterned Surfaces with Tailored Molecular and Surface Multivalency

<Valiokas R, Klenkar G, Tinazli A, Tamp R, Liedberg B, Piehler J (2006) Differential Protein Assembly on Micropatterned Surfaces with Tailored Molecular and Surface Multivalency. ChemBioChem 7, 1325 – 1329..>

Multivalent chelator head groups

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Multivalent chelator head groups

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Multivalent chelator head group

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Microstructured functional protein arrays

 

µ-contact printing combined with piezo-dispensing

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Thickness maps

Spotted bis-NTA density array (0, 1, 2, 5, 10, 20, 30 and 50 mol% in matrix)

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Immobilization of ifnar2-H10 and ligand binding

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Bis-NTA density array, SPR imaging

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mono- and bis-NTA density array

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mono- and bis-NTA density array

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mono- and bis-NTA density array

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 DNA – bar-coding of vesicles for bio chip application

<Klenkar G, Brian B, Ederth Th, Stengel G, Höök F, Piehler J, Liedberg B (2008) Biointerphases 3: 29-37.>

 

DNA – bar-coding of vesicles for bio chip application

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Time dependency, adsorption kinetics, biomolecular interaction

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 A microarray chip for label-free detection of narcotics 

<Klenkar G, Liedberg B (2008) Anal Bioanal Chem 391:1679–1688.>

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Characterization of physical properties of supported phospholipid membranes  

<HOWLAND MC, SZMODIS AW, SANII B, PARIKH AN. (2007) Characterization of physical properties of supported phospholipid membranes using imaging ellipsometry at optical wavelengths. Biophys J. 92, 1306-17>

 

Supported lipid bilayer

A lipid bilayer is a thin polar membrane made of two layers of lipid molecules. These membranes are flat sheets that form a continuous barrier around cells. The lipid bilayer is one of the most important self-assembled structures in nature.

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Solid supported lipid bilayers are an excellent model system for studying the surface chemistry of membranes and cell. A wide variety of surface-specific analytical techniques can be used to investigate processes such as cell signaling, ligand–receptor interactions or enzymatic reactions occurring at the cell surface

Photopattern of supported phospholipid membrane

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Biomolecular interaction at model membranes

Ganglioside GM1 interacting with cholera toxin B sub-units

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DPPC = 1,2-Dipalmitoyl-sn-Glycero-3-Phosphocholine

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Endpoint analysis via Imaging SPREE (high density protein arrays, >2500 spots, label-free, in situ) 

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Quality control of protein micro arrays

Quality control of protein spots (micro array)

 

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Ellipsometric contrast micrograph

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Delta map

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Delta map: digital zoom

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Delta map: profil

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Thickness map, sample A

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Thickness map, sample B

 

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new cells and accessories

nanofilm_microlab

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Instrumentation

The new imaging ellipsometer

Nanofilm_EP4

A modular platform allows us building the instrument according to your Scientific needs!

Start your experiences with imaging ellipsometry with the single wavelength version.

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Technical Integration with QCM-D from Q-Sense

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The new instrument offers a broad range of Unique features!

Accessories

Active vibration isolation halyconics_variobasic_40 Supportframe halcyonics_bam/ie

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Accessories imaging SPR in the ellipsometric mode

• KineticSPR cell

• Microfluidic

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Accessories in situ

• Various S/L cells

• Temperatur control

• Liquid handling

• Electrochemistry upgrades

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