Atomic force microscopy in histology and cytology.
This review briefly introduces the principles of the atomic force microscope (AFM) and shows our own results of AFM application to biological samples. The AFM, invented in 1986, is an instrument that traces the surface topography of the sample with a sharp probe while monitoring the interaction forces working between the probe and sample surface. Thus, the AFM provides three-dimensional surface images of the sample with high resolution. The advantage of the AFM for biologists is that AFM can visualize non-conductive materials in a non-vacuous (i.e., air or liquid) environment. AFM images of the plasmid DNA are comparable to those by transmission electron microscopy using a rotary shadowing technique, and have the advantage of examining directly the molecule without staining nor coating. The surface structure of human metaphase chromosomes and mouse collagen fibrils demonstrated in air by the non-contact mode AFM is comparable to that obtained by scanning electron microscopy. Quantitative information on the heights of structures is further obtainable from the AFM images. Embedment-free thin tissue-sections are useful for observing intracellular structures by AFM. The present review also shows AFM images of living cultured cells which have been collected in a contact mode in liquid. This technique afforded us three-dimensional observation of the cellular movement with high resolution. Although there are some innate limitations for AFM imaging, the AFM has great potential for providing valuable new information in histology and cytology.
Ushiki
T
T
Department of Anatomy, Niigata University School of Medicine, Japan.
Hitomi
J
J
Ogura
S
S
Umemoto
T
T
Shigeno
M
M
D000818:Animals; D002875:Chromosomes / Q000648:ultrastructure; D003094:Collagen / Q000648:ultrastructure; D003584:Cytological Techniques / Q000639:trends*; D004247:DNA / Q000648:ultrastructure; D006652:Histological Techniques / Q000639:trends*; D006801:Humans; D051379:Mice; D018625:Microscopy, Atomic Force / Q000295:instrumentation*
D003094:Collagen; D004247:DNA