BioScope Catalyst for High Resolution AFM Imaging of Biomolecules

April 20, 2011
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By AZoNano Editors

Table of Contents

Introduction
Atomic Force Microscopy
Nucleic Acids
Proteins
Membranes and Membrane Proteins
Conclusion
Bruker

Introduction

The BioScope Catalyst along with optical microscopy provides life science researchers an opportunity to study the biological species on a broad range of size scales. The BioScope Catalyst has advanced engineering and mechanical stability, hence high resolution three-dimensional images of single biomolecules and biomolecular complex structures can be obtained. The imaging conducted using the BioScope Catalyst provides data at single cell level by characterization into nucleic acids and proteins and membranes, etc.

Atomic Force Microscopy

The Atomic Force Microscopy (AFM) technique provides high-resolution imaging at nanoscale resolution of a three- dimensional structure without staining or coating the sample. Thus, AFM scores over many other techniques by allowing studies of biomolecules and parameters of biological processes at the process site itself and taking real time readings. The BioScope Catalyst AFM can be used with light microscopy techniques to provide optically guided navigation of the probe and creating clear images correlating the AFM and optical image data. Live cells were studied by using the BioScope Catalyst along with the largest closed loop, X-Y scan range; the results were highly accurate. The results are illustrated in Figure 1.

(A)

(B)

Figure 1. (A) Registered image overlay of a two-channel confocal laser scanning fluorescence microscopy image and AFM topography image of fibroblast cells labelled with Alexa Fluo 546 Phalloidin (red) and DAPI (blue). The BioScope Catalyst MIRO software enables registration of an optical field of view to the AFM scan area. Optical images can then be used to navigate the AFM probe to a region of interest to perform high resolution AFM imaging and/or highly sensitive force measurements. (B) Region of interest obtained from the image overlay showing correlated AFM and fluorescence data channels. Confocal fluorescence images were obtained with a Leica SP5 confocal system and using a 40x oil immersion objective. AFM images were obtained with a BioScope Catalyst operated in contact mode in buffer solution using MLCT AFM probes (k ~0.01N/m).

The advanced mechanical stability and engineering of the BioScope Catalyst makes it suitable to study biomolecular species also. It provides consistent results even when used with an inverted optical microscope, as shown in Figure 2.

Figure 2. A 1

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