Application of Atomic Force Microscopy to Observation of Marine Bacteria

The atomic force microscopy (AFM) is a recently developed, bench-top instrument that can image the surface structures of biological specimens at high resolution with simultaneous measurement of their size. This paper describes the application of AFM to marine bacteria. Both natural and cultured bacteria were retained on a filter or placed on glass, washed, air-dried and observed by AFM. The instrumental condition, the choice of suitable filter, effect of fixation and filtration, comparison with epifluorescent microscopic (EFM) count, and the size and shape of bacterial cells were investigated. An Isopore filter was best for concentration and subsequent observation because of its surface flatness. Cross section images showed that both rod and coccoid cells were flattened, the former usually having a two-humped shape. Bacterial cells were differentiated from non-living particles based on their cross section shape and size. Bacterial counts by AFM and EFM showed good agreement. Although size measurement is easily done by the instrument, AFM tends to overestimate the size of microspheres. More work is thus needed on the size measurement of living organisms. Because AFM easily provides images of natural bacterial cells at high magnification, it can be used as a new tool to study the fine structures of marine bacteria. This revised version was published online in July 2006 with corrections to the Cover Date.