Method for preparing thin sections of untreated equine hoof horn for electron microscopic examination.

The study explores a new method for preparing thin sections of the equine hoof horn for electron microscopic examination. The proposed technique promises to maintain tissue ultrastructure, save time and resources, and improve the quality of microscopic images.

Problem Statement

• The researchers highlight the difficulty in preparing hard tissues such as the equine hoof horn for electron microscopic examination.
• Issues relate to the penetration of fixatives and chemicals used during fixation and embedding, which often compromise the tissue’s ultrastructure and antigencity.

Objectives and Methodology

• The study aimed to identify and implement a new method that would enable a very high-quality preparation of thin sections of hoof horn and other hard tissues, with the condition of maintaining the tissue’s ultrastructure and suitability for immuno-labeling.
• This implied developing a method that differs from the commonly used fixation and embedding techniques, focusing more on using untreated materials for the preparation of these thin sections.

Findings and Benefits

• The researchers found that the new preparation method saves time and resources while providing equivalent ultrastructural information as the traditional methods.
• They also observed that thin sections from untreated material are significantly larger and more homogeneous. These sections exhibit high resistance to electron rays, hence are better suited for sectioning.
• Electron microscopic images obtained from this method can be compared to results achieved from fixed and embedded material, thus providing a clear picture of the state of the tissues.
• The method avoids fixation and embedding artifacts, thus enabling clearer interpretation of the electron microscopic findings.
• Significant gains were observed when immune-histochemical techniques were applied to untreated horn specimens, mainly because fixations invariably reduce a tissue’s antigenicity. Thus, the study found that with the avoidance of fixation, clearer and accurate observations could be made.