Low-cost educational stochastic optical reconstruction microscopy (eSTORM)

Abstract

A stochastic optical reconstruction microscope was built and demonstrated for under $20,000, enabling hands-on learning of single-molecule localization concepts in teaching laboratories. This was accomplished by replacing the most expensive parts of $500,000 commercial instruments, namely the laser, camera, and objective, with lower cost alternatives. Since lower cost also comes with higher noise, we characterized the optical and noise characteristics of the microscope. A new sample protocol, consisting of microspheres labeled with streptavidin-Alexa 647 conjugates, was developed to test the system, compare the image quality of two reconstruction programs (QuickPALM and rapidSTORM), and evaluate trade-offs in camera selection. Finally, by imaging defined actin features in 3T3 cells, the instrument was estimated to have a sub-diffraction resolution between 70 -100 nm.