C. Jehn , B. Küstner, F. Schöppler, F. Baum, M. Schütz, M. Gellner and S. Schlücker Institute of Physical Chemistry University of Würzburg Germany http://www.phys-chemie.uni-wuerzburg.de SERS Microscopy (µSERS) Selective and Sensitive Localization of Proteins in Tissue Specimens SERS Round Table Discussion, Poltersdorf, October 3-5, 2007
22
Embed
C. Jehn , B. Küstner, F. Schöppler, F. Baum, M. Schütz, M. Gellner and S. Schlücker
SERS Microscopy (µSERS) Selective and Sensitive Localization of Proteins in Tissue Specimens. C. Jehn , B. Küstner, F. Schöppler, F. Baum, M. Schütz, M. Gellner and S. Schlücker Institute of Physical Chemistry University of Würzburg Germany. http://www.phys-chemie.uni-wuerzburg.de. - PowerPoint PPT Presentation
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
C. Jehn, B. Küstner, F. Schöppler, F. Baum, M. Schütz, M. Gellner and S. Schlücker
Institute of Physical ChemistryUniversity of Würzburg
Germany
http://www.phys-chemie.uni-wuerzburg.de
SERS Microscopy (µSERS)
Selective and Sensitive Localization
of Proteins in Tissue Specimens
SERS Round Table Discussion, Poltersdorf, October 3-5, 2007
Outline
Immunohistochemistry: Basics
Biofunctionalized Nanoparticles
SERS Microscopy: Proof of Principle
Novel SERS labels
SERS Imaging of PSA in Prostate Biopsies
Outlook
Label
1
Immunohistochemistry
Tissue Section
+1. Label
2. Remove Unbound Antibodies
2
Protein
Protein
Target Protein/Antigen
Tissue Section
Label
Label
Multiplexing Capacity of Different Labelling Approaches
Fluorescence Emission
Fluorophores Quantum Dots
ca. 1-3 ca. 3-10
Absorption
Dyes
ca. 1-3
Raman Scattering
SERS Labels
ca. 10-100
Immunohistochemistry (IHC): Quo Vadis ?
3
• small Raman line width• minimal spectral overlap
Functionalized Nanoparticles for Nanodiagnostics
+ +
Immuno-SERS Label
AntibodyGold
NanoparticleRaman Label
Spacer +
Spacer
4
Self-Assembled Monolayer (SAM) on Gold Nanoparticles
Spacer
5
Hollow Gold Nanoparticles: Synthesis
Ag colloid
max ~ 400 nm
Au-Ag alloy
max 550 – 900 nm
Nanoparticles with tunable surface plasmon resonance !
3 Ag + Au3+ 3 Ag+ + Au
Solid Ag spheres Hollow Au shells
6
Hollow Gold Nanoparticles: Characterization
400 500 600 700 800 900 1000 1100
Ab
sorb
an
ce
Wavelength/nm
78
5
72
5
63
0
83
0
Plasmon resonance energy depends on shell
thickness
50 nm
7
Wavenumber / cm-1
Ra
ma
n In
ten
sity
1750 1500 1000 750 1250
Spectrum of Immuno-SERS lable
Selected Raman bands:
• SERS distance dependence
• SERS selection rules
• Electronic resonance
8
D. S. Grubisha, R. J. Lipert, H.-Y. Park, J. Driskell, M. D. Porter, Anal. Chem. 2003, 75, 5936.