High-throughput multiplex bioelectric clamp profiling of acutely isolated blastemal progenitors
June 26
The whole-cell microelectrode clamp remains the gold-standard for interrogating excitable embryonic and regenerative cells. Nonetheless, its conventional execution is labor-intensive and inherently low throughput. Although recently introduced robotic clamp platforms have accelerated compound screening, they have largely centered on heterologous systems overexpressing single ion channels or receptors.
We have established an automated, high-throughput bioelectric clamp strategy that enables simultaneous, unbiased characterization of acutely dissociated blastemal progenitors in their native physiological state. To curate and interrogate the resulting large-scale datasets, we created open-source software featuring an intuitive graphical interface that fits each cell’s currents with biophysical models, thereby generating a functional fingerprint for every individual progenitor.
Here, we delineate a streamlined protocol encompassing: (1) enzymatic and mechanical dissociation of progenitor populations from regenerating tissue; (2) design and execution of multiplex clamp recordings on the robotic platform; and (3) automated analysis employing predefined, unbiased filtering parameters. The methodology is applicable to diverse objectives, from mapping developmental bioelectric profiles to high-content screening for pro-regenerative therapeutics. The complete workflow—cell preparation, data acquisition and computational analysis—requires 6–18 h. Practitioners should possess graduate-level proficiency in tissue dissection, electrophysiology and biophysical modeling.