Basic NeuroscienceA time and cost efficient approach to functional and structural assessment of living neuronal tissue

In this manuscript, we describe a protocol for capturing both physiological and structural properties of living neuronal tissue. An essential aspect of this method is its flexibility and fast turnaround time. It is a streamlined process that includes recording of electrophysiological neuronal activity, calcium imaging, and structural analysis. This is accomplished by placing intact tissue on a modified Millicell Biopore insert. The Biopore membrane suspends the tissue in the perfusion solution, allowing for complete access to nutrients, oxygen, and pharmacological agents. The ring that holds the membrane ensures its structural stability; forceps can be used to grip the ring without contacting the filter or the tissue, for easy transfer between multiple setups. We show that tissue readily adheres to the surface of the membrane, its entire surface is visible in transmitted light and accessible for recording and imaging, and remains responsive to physiological stimuli for extended periods of time.

► The described technique allows for an affordable and fast comprehensive physiological, anatomical, and immunological characterization of living neuronal tissue. ► Intact tissue is placed on a modified Millicell Biopore insert, which reduces physical damage to the tissue, maintains its structural integrity, and allows easy transfer across multiple setups. ► The approach uses affordable alternatives to common techniques to achieve the same results.