BCG cell imaging using scanning probe microscopy.

Abstract

Scanning tunneling microscopy (STM) and atomic force microscopy (AFM) were used to obtain images of the surface of whole, intact BCG (bacille Calmette Guerin, a mycobacterium) cells in air and under solution by immobilizing the cells onto glass slides (AFM only) or highly oriented pyrolytic graphite. The technique used for AFM imaging involved depositing a submonolayer of cells under a centrifugal force followed by fixation/dehydration using polar organic solvents. AFM images agree well with images from light and electron microscopy and showed large numbers of BCG cells in their distinctive cord arrangement. The AFM also proved useful for identifying extracellular microgranules which cannot be seen with light microscopy. For STM imaging, the hydrophobicity of BCG enabled strong adhesion from aqueous solution onto graphite. STM images of BCG could only be obtained while scanning in aqueous solution, and the cells showed a large variation in contrast when different samples were imaged. The STM provided greater detail of surface features than the AFM and was able to produce images of periodic layers corroborating observations made by transmission electron microscopy.