Atomic force microscopy of histological sections using a chemical etching method.
Physiology and pathology have a big deal on tissue morphology, and the intrinsic spatial resolution of an atomic force microscope (AFM) is able to observe ultrastructural details. In order to investigate cellular and subcellular structures in histological sections with the AFM, we used a new simple method for sample preparation, i.e. chemical etching of semithin sections from epoxy resin-embedded specimens: such treatment appears to melt the upper layers of the embedding resin; thus, removing the superficial roughness caused by the edge of the microtome knife and bringing into high relief the biological structures hidden in the bulk. Consecutive ultrathin sections embedded in epoxy resin were observed with a transmission electron microscope (TEM) to compare the different imaging properties on the same specimen sample. In this paper we report, as an example, our AFM and TEM images of two different tissue specimens, rat pancreas and skeletal muscle fibres, showing that most of the inner details are visible with the AFM. These results suggest that chemical etching of histological sections may be a simple, fast and cost-effective method for AFM imaging with ultrastructural resolution.
Tiribilli
B
B
Biophotonics Laboratory, National Institute of Applied Optics, Florence, Italy. tiribilli@inoa.it
Bani
D
D
Quercioli
F
F
Ghirelli
A
A
Vassalli
M
M
D000818:Animals; D004853:Epoxy Resins; D000432:Methanol; D018625:Microscopy, Atomic Force; D046529:Microscopy, Electron, Transmission; D018482:Muscle, Skeletal / Q000648:ultrastructure; D010179:Pancreas / Q000648:ultrastructure; D051381:Rats; D013048:Specimen Handling / Q000379:methods*
D004853:Epoxy Resins; D000432:Methanol