3D culture of isolated cells: a fast and efficient method for optimizing their histochemical and immunocytochemical analyses

The rapid development of three-dimensional (3D) culture systems and engineered cell-based tissue models gave rise to an increasing need of new techniques, allowing the microscopic observation of cell behavior/morphology in tissue-like structures, as clearly signalled by several authors during the last decennium. With samples consisting of small aggregates of isolated cells grown in suspension, it is often difficult to produce an optimal embedded preparation that can be further successfully processed for classical histochemical investigations. In this work, we describe a new, easy to use, efficient method that enables to embed an enriched "preparation" of isolated cells/small 3D cell aggregates, without any cell stress or damage. As for after tissue-embedding procedures, the cellular blocks can be further suitably processed for efficient histochemical as well as immunohistochemical analyses, rendering more informative-and attractive-studies onto 3D cell-based culture of neo-tissues. Microsc. Res. Tech., 2015. © 2015 Wiley Periodicals, Inc.