AFM combined with inverted optical microscope
Combined imaging using an inverted optical microscope and AFM offers a unique opportunity to correlate optical data (such as phase contrast or fluorescence data) with high-resolution topography and mechanical data that only AFM can provide.
Thanks to the open optical path design of the FlexAFM, light can pass through the FlexAFM with minimal obstruction. Additionally, the special SureAlign™ optical design of the cantilever holder SA simplifies FlexAFM operation in liquid environments and eliminates the need for laser alignment. When integrated with the inverted optical microscope option, the FlexAFM becomes an ideal system for imaging and characterizing cells under physiological conditions. In this example, live Rat-2 fibroblasts were imaged.
Applications of FlexAFM combined with inverted optical microscopy are by no means limited to life sciences. Presented below is an analysis of a coated glass sample using optical microscopy and atomic force microscopy. AFM delivers higher-resolution data across the imaged area and reveals the nature of structures observed under the optical microscope. It can also be used for coating thickness measurement.
Images of cells in a Petri dish: (Left) Phase contrast image from the inverted optical microscope acquired with a FlexAFM unit and inverted optical microscope option, showing live Rat-2 fibroblasts and the AFM cantilever in cell culture medium. (Right) AFM image of the cells captured with the same setup, displaying fine details of the Rat-2 cytoskeleton, which can be readily distinguished through the cell membrane. AFM image size: 70 μm × 60 μm.
Phase contrast image (top) and AFM image (bottom) of coated glass. The pore structures within the coating, corresponding to textured features (e.g., inside elliptical regions) seen under the optical microscope, are verified by AFM measurement. They provide a reliable method to determine coating thickness, which in this example is 40 nm. AFM image size: 90 μm × 35 μm.
