Monkey's Kidney

Monkey Kidney Cells 
Fluorescence NSOM Images
  
 

     
35x35 micron AFM image    35x35 micron Transmission FluorescenceNSOM image

These images were produced using in the Intermittant Contact mode in a liquid environment using the Nanonics Liquid Cell with the MultiView 1000 system.

 

These topographic and fluorescence images were obtained simultaneously with cantilevered NSOM fiber optic probes. Labelled with rabbit anti-fibronectin primary antibody which was followed by a goat anti-rabbit Alexflour 488 secondary antibody. The Alexaflour probe is excited at about 488 and emits approximately 520nm. Thus, all fibronectin should be fluorescent. Fibronectin is part of the extracellular matrix which attaches the cell to the glass substrate.

The cell is visible in the AFM topography image but the fluorescent image shows mainly fibers. The fibers are primarily under the cell and therefore are difficult to image with AFM unless the cell is thin enough and these cells are thick since they were imaged at a more advanced stage in their growth. However, the fibers are fluorescent so they show up in the fluorescence NSOM optical image.