Definition of a sectioning plane and place for a section containing hoped-for regions using a spare counterpart specimen

Histological examination of targets in regions of interest in histological sections is one of the most frequently used tools in biomedical research. However, it is a technical challenge to secure a multitarget section for inspection of the structure’s mutual relationship of targets or a longitudinally filamentous- or tubular-formed tissue section for visitation of the overall morphological features. We present a method with a specified cutting plane and place, allowing researchers to cut directly at the multitarget centers accurately and quickly. The method is proven to be reliable with high accuracy and reproducibility and a low coefficient of variation, testing on repeat experiments of three target’s position-known models. With this method, we successfully yielded single sections containing whole intraorbital optical nerves, three aortic valves, or whole thoracic tracheas in their central positions. The adjoined custom-made tools used in the study, such as various tissue-specific formulated calibrated trimming and embedding guides, an organ-shaped cavity plaster mold, and a two-time embedding technique for optimal and identical trimming or embedding, also bear great potential to become a common supplemental tool for traditional histology and may contribute to the reduction of the labor, and the number of animals needed.