Design and evaluation of low-cost, DIY programmable tissue processor for solvent exchange in biological sample preparation.

Abstract

Imaging techniques are fundamental tools in biology for examining cell growth and responses to the environment. Many tissues require fixing, staining, and/or clearing before they can be visualized under a microscope. However, these protocols, such as those using propidium iodide (PI), a fluorescent cationic stain widely used across biological specimens including plant, mammalian, and bacterial, often require laborious dehydration and rehydration steps to facilitate stain penetration. These stepwise solvent exchanges, for example, by passing tissues through a graded ethanol series, are time-consuming and manually intensive. While automated tissue processors offer an alternative, they are outside of the budget for many labs. Here, we present an open-source, low-cost (~$400) automated tissue processor that performs sequential dehydration and rehydration of biological tissues, significantly reducing hands-on labor. The processor is made of readily available, standardized parts and includes custom software that allows users to define and save protocols. We demonstrate the use of the processor by automating a multi-day PI staining protocol across multiple plant species, tissue morphologies, and users, and by comparing tissue quality with hand-processed samples. Our design provides a low-cost, accessible alternative to expensive commercial tissue processors, offering a practical solution for a wide range of biology laboratories.