AFM-NSOM Measurements of Healthy Human

Dermal Fibroblasts in Liquid

A demonstration of Nanonics' unique abilities in both biological and soft sample imaging.

The imaging of soft tissues, such as the live fibroblast cells shown below, leads the way to many new biological applications. The human dermal cells were imaged live in a PBS buffer (pH-7.3).  Below we see a chain of NSOM images obtained in transmission mode with a Nanonics patented cantilevered tip with  an aperture diameter of 100nm.

50 x 50 microns	30 x 30 microns	8 x 8 microns

These images illustrate  the abilities of Nanonics' MultiView systems to overcome the difficulties usually associated with viewing live, soft tissue liquid samples. These SPM systems permit easy viewing of liquid samples via a versatile and  user-friendly liquid cell attachment. The open architecture of Nanonics' SPM heads allows one to view samples from above and below. These systems are easily integrated into our dual microscope which combines both upright and inverted formats. This is particularly important for better resolution of low-contrast biological samples.

The free optical axes also allows fluorescence or Raman imaging to be accomplished with ease.

 Using the same system AFM was used for topographical mapping. The AFM was carried out in intermittent contact mode, as regular contact mode would have scratched the soft tissue. Nanonics systems are able to operate in all three SPM modes.

CCD sample image with NSOM probe approaching  the surface

Below we see the AFM images (left) and the corresponing (right) NSOM images.

Below is shown a line scan of simultaneously obtained topography and NSOM images. The AFM image is at the top and the NSOM image is at the bottom.
  

Human dermal fibroblasts are derived from the dermis of normal human skin and have been used for in vitro analyses of fibroblast growth, migration and collagen metabolism in wound healing. Fibroblasts are responsible for the homeostasis of connective tissues and their activity is modulated by several cells and soluble factors. SPM techniques using our MultiView systems were used as part of a study to investigate the factors that regulate the synthesis of the extracellular matrix molecules and the functional organization of these molecules in the extracellular space.