Cryoprocessing, upgraded: A scalable solution for tissue and organoid research

Understanding the molecular landscape of cells is key to studying health and disease, with biomarkers—measurable indicators of biological activity—playing a central role in both research and diagnostics. Identifying such cellular biomarkers is, for example, essential for advancing both fundamental research and critical diagnostic applications, such as analyzing biopsies for potential cancer markers. Traditionally, tissue sectioning, which produces ultra-thin slices for downstream analysis, has been the gold standard for assessing protein and RNA expression through techniques like immunohistochemistry and in situ hybridization. However, a key limitation of immunohistochemistry is time and cost, which restrict its broader use.   

Now, Daniel Reumann and Jürgen Knoblich at the Institute of Molecular Biotechnology (IMBA) of the Austrian Academy of Sciences unveil a novel technique in which newly developed multiplexed tissue molds (MTMs) are used to process over 100 tissue specimens simultaneously. Published on April 21 in Cell Reports Methods, this method reduces processing and analysis costs by as much as 96%, while minimizing the ecological footprint of sample processing and storage: MTMs are re-usable, save reagents and drastically reduce the need for freezer capacity.

With the growing demand for large-scale organoid and 3D tissue studies, ensuring that experimental conditions remain uniform across all samples is more critical than ever. Even small experimental steps can introduce heterogeneity into the experiment; therefore, it is crucially important to process samples simultaneously under the same conditions. In a proof-of-principle study, the researchers successfully processed an array of 19 heterogeneous mouse tissues ranging from soft to hard and conducted a time-course experiment monitoring protein expression during neurogenesis in over 100 neural organoids. This analysis would have cost well above 1.000€ in reagents with conventional methodology. Employing the new processing technique using MTMs, the researchers brought the costs down to less than 50€. Furthermore, the simultaneous processing minimizes variability and paves the way for more reproducible immunohistochemical analyses in both research and clinical diagnostics.

By streamlining immunohistochemical workflows and ensuring remarkable reproducibility across heterogeneous samples, this technology paves the way for transformative advances in both research and clinical diagnostics - and enables diagnostics where cost is a prohibitive factor.

“This scalable solution drastically reduces cost and labor, helping research and diagnostic labs to more easily use immunohistochemistry as a readout. At the same time, our solution maintains tissue integrity and improves consistency”, says Daniel Reumann, previously a PhD student in the lab of Jürgen Knoblich at IMBA and now postdoc at the Kavli Institute of Nanoscience Discovery at the University of Oxford. Jürgen Knoblich, deputy scientific director at IMBA, adds: “This new method enables researchers to analyze tissue samples at a large scale, while enhancing the reproducibility of experiments.”