AFM for Life Sciences Applications

Atomic force microscopes are capable of making measurements on biological samples that are difficult or impossible with any other type of microscope. Some examples of life sciences applications unique to atomic force microscopes are shown below.

Measuring images of biomolecules in ambient air and liquids

Atomic force microscopes are the only microscopes capable of imaging bio-molecules in ambient air and liquids.

Double-stranded DNA Molecules

Life sciences imaging with AFM, DNA 5 µm x 5 µm

5 µm x 5 µm image of double-stranded DNA molecules.

Imaging of oligonucleotides such as these can be used in a wide variety of applications, notably in structural studies, and in the study of interactions between the oligonucleotides and other molecules, such as enzymes.

 

Tobacco Mosaic Virus (TMV)

AFM for Life Sciences, Tobacco Mosaic Virus, 1.2 µm x 1.2 µm

1.2 µm x 1.2 µm image of tobacco mosaic virus (TMV).

Imaging of viruses is commonly carried out in structural studies. In addition, TMV is commonly used as an imaging standard since it has a highly conserved structure. Imaging high aspect ratio samples such as these also helps to characterize tip sharpness.

 

Such high resolution imaging requires an instrument noise floor of less than 0.1nm, a good tip approach leading to the maintenance of a sharp tip, the capability to scan with very light forces, and placement of the microscope in an environment with minimal structural and acoustic vibrations.

Proper sample preparation is another critical factor. Sample preparation techniques are described in Atomic Force Microscopy by Eaton and West.

Imaging cells in ambient air or liquid

Atomic force microscopy has a particular advantage over electron microscopy, in that cells can be imaged in partially or totally hydrated conditions.

Parasites

AFM for Life Sciences, 25 µm x 25µm, parasites

25 µm x 25µm image of parasites measured in air.

These Leishmania cells have been treated with an antimicrobial peptide, leading to highly roughened cell membranes, which can be measured and quantified by AFM.

 

Epithelial Cells

AFM scan, epitheleal cell measured in liquid 32 µm x 32 µm

32 µm x 32 µm image of epithelial cells measured in liquid with AFMWorkshop's Dunk and Scan scanner.

Measurement of high resolution images of cells in liquid (e.g., under physiological conditions) is another possibility unique to AFM, and can give much more relevant results than electron microscopy, which requires cell fixation, leading to artifacts.

 

Bacteria Spore Mutants

30 µm x 30 µm AFM scan of bacteria spores

30 µmx 30 µm image of bacteria spore mutants.

The ability to image a very large number of cells, such as these spores, allows the researcher to obtain statistically relevant information about a population of cells. Images of multiple cells can be also useful to assess inter-cellular effects, such as clustering and adhesion.

 Imaging cells in combination with an inverted optical microscope

The inverted optical microscope facilitates direct placement of the probe on an area of interest for scanning. Additionally the inverted microscope can be operated in epiflourescence mode.

Neutrophil A Cells

Life Sciences inverted optical microscope image of neutrophil -A cells

Inverted optical microscope image of neutrophil A cells. The dotted outline is the area scanned with the AFM.

Light shaded AFM image neutrophil-A cells

Light Shaded AFM image of the cells visualized in the optical microscope image.

 

Caco-2

Life Sciences AFM Caco-2 cells

Inverted optical microscope image of Caco-2 cells in the LS-AFM.

Clearly visible is the AFM cantilever on the right side of the image. A box identifies the area for AFM scanning.

3-D scale of Caco-2 cell 48 µm x 48 µm

3-D color scale image of the Caco-2 cell.

The scan range is 48 µm x 48 µm.

 

Epifluorescence and topographic AFM image of Caco-2 and quantum dots

CACO-2 cell structure in the presence of low concentration of quantum dots.

Left: Epifluorescence, showing brightfield (red), DAPI (blue), 2.2nm quantum dot PL emission at 560nm (green).

Right: Topographic AFM image of the indicated area.

Measuring the stiffness of biomaterials at the nano scale

Monitoring the deflection of a cantilever as it is pushed against a sample results in a force/distance curve. From the force distance curve many parameters may be measured, such as stiffness of the sample and probe-sample adhesion.

In biological samples, the most common application is measurement of intermolecular forces. For example, this could be used to measure the interaction force between an antigen and an antibody directly. Cell-cell adhesion forces and cellular stiffness can also be measured.

Advanced Force Distance Curve AFM software

The above screen shot demonstrates Advanced Force Distance Curve software measuring an AFM image.

1. Force-Distance data display region
2. Slider indicates the extension of the Z piezoelectric ceramic
3. Control parameter selection options
4. AFM Image for selecting locations for force-distance measurements

Recommended AFM products for life sciences applications:

TT-Atomic Force Microscope ‒ Measurements of biomolecules and biomaterials

LS-Atomic Force Microscope ‒ Imaging cells and measuring stiffness/adhesion