Generation of Three-dimensional Printed Biological Innovative Mold for Optically Transparent Tissue Specimens

This article has been accepted and is currently in production

Abstract

Histology continues to evolve in modern day experimentation. Recently, groups have been able to make tissue and bones transparent by removing lipids and other biological species that prevent deep tissue imaging. Now, groups can robustly "clear" the tissue or specimen and perform a wide array of microscopy such as single-photon, multiphoton, and light-sheet to evaluate their biological target of interest. This has eliminated the need to freeze, cyro-slice, and cyro-preserve tissue, greatly limiting human prone error. However, current tissue clearing methods still lack a robust system to mount and structurally support unique specimens. This leads to issues with tissue movement during imaging, the lack of reliability when re-staining and re-evaluating specimens, and working with abnormally shaped specimens such as tumors. To overcome these obstacles, the innovative Mold (iMold) was generated. iMolds are first created from images of the specimen followed by three-dimensional (3D) printed molds that support the unique tissue structure being imaged and re-imaged. This leads to a dramatic reduction in tissue movement while imaging and allows for scientists to robustly re-evaluate areas of interest that have been re-stained or need to be re-imaged. Furthermore, iMolds can be created in a short time period and generated for multiple samples in any imaging format (i.e. slide, cell culture plate). This allows users to scan multiple organs or specimens on one slide or cell culture dish. In closing, iMolds can be utilized by any research laboratory studying cleared specimens while also reducing costs, time, and tissue movement compared to other current methodologies.