Nanosonden: Neue Konzepte und Anwendungen in der Biomedizin

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1 Nanosonden: Neue Konzepte und Anwendungen in der Biomedizin Harald Fuchs Physikalisches Institut Westfälische Wilhelms-Universität Münster & Center for Nanotechnology (CeNTech) Germany Münster,

Wilhelms-Universität Münster & Center for Nanotechnology

2 Inhalt : Einleitung: Nanoskopie von Zellen und Gewebe Rastersonden: AFM - Technische Entwicklungen - AFM auf biologischen Oberflächen DIP-Pen Lithographie für biofunktionale Oberflächen

3 Komplementäre Nano-Bioanalytikmethoden - LM (FM, CFLM, 4 π, STED, PALM.. ) - Massen Spektrometrie (TOF-SIMS) - Elektronenmikroskopie (TEM/CTEM) - Elektrische Charakterisierung (CellZscope) - AFM, SICM, SNOM

4 Ziel: Einzelzellanalytik für die Diagnose Diagnostics Test of Therapeutics Drug Development Gastroenterologische Molekulare Zellbiolog Med. Klinik und Poliklinik B

5 Nano - Analytik Mit kleinsten Stoffmengen Hochauflösende MS

6 Time-of-Flight Mass Spectrometers Time-of-Flight Secondary Ion Mass Spectrometry Chemical Surface Analysis ION-TOF GmbH Heisenbergstr. 15 D Münster / Germany

7 Distribution of 10B by Laser-SNMS in kidney tissue 10 B CN C 3 H 8 N CN-C 3 H 8 N- 10 B-Overlay Red: CN CN-C 3 H 8 N-Overlay Green: C 3 H 8 N Blue: 10 B Area: 120 x 120 µm 2 Prof. Dr. H. Arlinghaus, Münster

8 Weiterentwickelte Lichtmikroskopie

9 4Pi-Microscopy Prof. R. Peters, CeNTech

10 Cryo - TEM

11 Elektrische Charakterisierung von Zellschichten

12 Measuring the electrical impedance of epithelial and endothelial cell layers under physiological conditions

13 Transepithelial electrical resistance (TER) of primary cultured endothelial cells derived from porcine brain microvessels, incubated in serum-free medium supplemented with hydrocortisone and without hydrocortisone adapted from Wegener et al., BioTechniques 37, 590 (2004)

incubated in serum-free medium supplemented with hydrocortisone and

14 cellzscope automated cell monitoring device for further informations:

15 Microscopes for the Nanoworld

16 Scanning Tunneling Microscopy (STM) STM (1981), G. Binnig, H. Rohrer, IBM Zürich

17 Scanning Nearfield Microscopy STM AFM SNOM SICM Raman-Spectr.

18 Atomic Resolution on Insulators with NC-AFM NaCl in UHV at RT Cl - Na + a = 0.56 nm 5 nm Δf = -140 Hz Schirmeisen, Weiner, Fuchs, Phys. Rev. Lett. 97, (2006)

19 Potential Energy Map ΔE barrier = 48 mev Schirmeisen, Weiner, Fuchs, Phys. Rev. Lett. 97, (2006)

20 PTCDA Multilayer Dynamic AFM Univ. Münster, 2006

21 CE - mode in air/liquids

22 QFM-Module for Applications in Air and Liquid BMBF-Project (Grant No. 03N8704) Junior Researchers Competition Nanotechnology Hendrik Hölscher Heisenbergstraße Münster, Germany

23 Contour Map of DPPC Monolayer in Air J.E. Schmutz, H. Hölscher et al., Ultramicroscopy (2007)

24 III Molekulare Elastiziät

25 Fast imaging of digestion process 3:48 (min:sec) 13:40 22:02 MPa :30 63:04 85:08

26 Applications in Neurology

27 Anatomy of a peripheral axon Schwann cell axon myelin myelin 1-10 µm

28 Myelinated nerve fibrils axon node of Ranvier myelin sheath plasma membrane of Schwann cell topography elastic modulus myelinated fibril demyelinated fibril myelinated fibril (force map)

29 Stiffness depends on ion concentration

30 Scanning Ion Conductance Microscopy

31 Living cells with SICM fresh MDCK II cells dying MDCK II cells 30µm x 30µm 100µm x 100µm

32 Rapid prototyping of multicomponent phospholipid nanostructure arrays using dip-pen nanolithography Steven Lenhert Institute für NanoTechnologie Forschungszentrum Karlsruhe, Germany

33 1. Background and motivation Demers, L. M.; Ginger, D. S.; Park, S. J.; Li, Z.; Chung, S. W.; Mirkin, C. A., Direct patterning of modified oligonucleotides on metals and insulators by dip-pen nanolithography. Science 2002, 296, (5574), Vega, Rafael A.; Shen, C. K. F.; Maspoch, D.; Robach, Jessica G.; Lamb, Robert A.; Mirkin, Chad A., Monitoring Single-Cell Infectivity from Virus-Particle Nanoarrays Fabricated by Parallel Dip-Pen Nanolithography. Small 2007, 3, (9), DNA Protein Phospholipids Lenhert, S.; Sun, P.; Wang, Y. H.; Fuchs, H.; Mirkin, C. A., Massively parallel dip-pen nanolithography of heterogeneous supported phospholipid multilayer patterns. Small 2007, 3, (1),

34 5. Biomimetic membrane systems His-tagged protein DPN patterns with nickel chelating lipids His Ni His Ni His Ni Biotinylated protein Streptavidin DPN patterns with biotinylated lipids Substrate (glass) Carrier (DOPC) Nickel chelating headgroup Biotinylated headgroup

36 Thanks to: B. Anczykowski L.F. Chi U. Fischer H. Hölscher S. Lenhert K. Riehemann T. Schäffer nanoanalytics

37 Herzlichen Dank für Ihre Aufmerksamkeit!

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