Background: One of the limitations when establishing an electrophysiology setup, particularly in low resource settings, is the high cost of microscopes. The average cost for a microscope equipped with the optics for infrared (IR) contrast or microfluorometry is $40,000. We hypothesized that optical elements and features included in commercial microscopes are not necessary to IR video-visualize neurons or for microfluorometry.
New method: We present instructions for building a low-cost epifluorescence upright microscope suitable for visualized patch-clamp recording and fluorescence detection using mostly catalog-available parts.
Results: This microscope supports applications such as visualized whole-cell recording using IR oblique illumination (IR-OI), or more complex applications such as microfluorometry using a photodiode. In both IR-OI and fluorescence, actual resolution measured with 2-μm latex beads is close to theoretical resolution. The lack of movable parts to switch configurations ensures stability when doing intracellular recording.
Comparison with existing methods: The low cost is a significant advantage of this microscope compared to existent custom-built microscopes. The cost of the simplest configuration with IR-OI is ∼$2000, whereas the cost of the configuration with epifluorescence is ∼$5000. Since this design does not use pieces discarded from commercial microscopes, it is completely reproducible.
Conclusions: We suggest that this microscope is a viable alternative for doing in vitro electrophysiology and microfluorometry in low-resource settings. Characteristics such as an open box design, easy assembly, and low-cost make this microscope a useful instrument for science education and teaching for topics such as optics, biology, neuroscience, and for scientific "hands-on" workshops.
Keywords: Calcium imaging; DIC IR; Fluo-4; Fluorescence; IR-OI; Microfluorometry; Visualized whole-cell recording.
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