An integrated instrumental setup for the combination of atomic force microscopy with optical spectroscopy.
In recent years, the study of single biomolecules using fluorescence microscopy and atomic force microscopy (AFM) techniques has resulted in a plethora of new information regarding the physics underlying these complex biological systems. It is especially advantageous to be able to measure the optical, topographical, and mechanical properties of single molecules simultaneously. Here an AFM is used that is especially designed for integration with an inverted optical microscope and that has a near-infrared light source (850 nm) to eliminate interference between the optical experiment and the AFM operation. The Tip Assisted Optics (TAO) system consists of an additional 100 x 100-microm(2) X-Y scanner for the sample, which can be independently and simultaneously used with the AFM scanner. This allows the offset to be removed between the confocal optical image obtained with the sample scanner and the simultaneously acquired AFM topography image. The tip can be positioned exactly into the optical focus while the user can still navigate within the AFM image for imaging or manipulation of the sample. Thus the tip-enhancement effect can be maximized and it becomes possible to perform single molecule manipulation experiments within the focus of a confocal optical image. Here this is applied to simultaneous measurement of single quantum dot fluorescence and topography with high spatial resolution.
Owen
R J
RJ
JPK Instruments AG, Bouchestrasse 12, 12435 Berlin, Germany. owen@jpk.com
Heyes
C D
CD
Knebel
D
D
Röcker
C
C
Nienhaus
G U
GU
D007091:Image Processing, Computer-Assisted / Q000295:instrumentation / Q000379:methods; D018625:Microscopy, Atomic Force / Q000295:instrumentation* / Q000379:methods; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D010784:Photomicrography / Q000295:instrumentation / Q000379:methods; D045663:Quantum Dots; D015203:Reproducibility of Results; D013057:Spectrum Analysis / Q000295:instrumentation* / Q000379:methods