Simultaneous brighfield, fluorescence and
AFM measurement
of epithelial cell.

Simultaneous AFM and Fluorescence
Imaging with the MFP-3D

Combining Atomic Force Microscopy (AFM) and fluorescence microscopy (FM) has long been of interest for biologists. These two types of microscopies individually provide unique information about samples. Fluorescence microscopy is typically used to detect specific molecules. However, this microscopy technique frequently has a number of drawbacks including limited spatial resolution and complex sample preparation.

Another limitation of this microscopy is that the molecule must be bound to the fluorochrome, which in turn, can alter the state of the molecule. On the other hand, AFM is a measurement made with a cantilever that is scanned cross the surface. It provides three dimensional molecular resolution of biological molecules with no additional contrasting agent (such as fluorochrome, dyes, etc.) or compromise to the sample.

In addition, AFM is also a higher resolution measurement that is non-fleeting. Using these microscopy techniques simultaneously provides the user with high resolution imaging on specific molecules and also opens the door to a wide variety of research experiments. In this application note, we will describe the instrumentation and setup, sample preparation, applications, and give experimental examples of the technique. For the full text, download a PDF copy of the AFM/Fluorescence Application Note.

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MFP-3D-IO on a Nikon TE 2000.