Ribbons of semithin sections: an advanced method with a new type of diamond knife

Abstract

Complete series of semithin sections are imperative for 3-D reconstruction, but with traditional microtomy techniques it is difficult and time-consuming to trace stained and labeled structures. In the present study we introduce a method for making and collecting ribbons of semithin sections with a new, commercial available diamond knife (histo-jumbo-diamond knife, Diatome AG, Biel, Switzerland). The special feature of the diamond knife is the large water bath (boat) into which a glass slide can be dipped. The method has distinct advantages and the handling is simple. The resin block is trimmed into a truncated pyramid. Contact glue is applied to the leading face of the pyramid, which makes sections stick together to form a ribbon. Following sectioning, the ribbons are mounted onto glass slides and aligned in parallel. Stretching out and drying the ribbons on a hot plate is the final step of the method. Major advantages of this method are the perfect alignment of sections with identical orientation of structures, the completeness of series, and the significant saving of time. This facilitates tracing of stained and labeled structures, yielding quick 3-D reconstruction. Semithin sections can be cut from 0.5 to 2 micro m and several ribbons can be mounted side by side onto the slide. Two examples are presented to illustrate the advantages of the method.