Simple buffers for 3D STORM microscopy

Authors:

Olivier, N., Keller, D., Rajan, V. S., Gönczy, P., & Manley, S.

Keywords:

High numerical aperture optics, Laser dyes, Structured illumination microscopy, Super resolution microscopy, Three dimensional imaging, Three dimensional microscopy

Abstract:

3D STORM is one of the leading methods for super-resolution imaging, with resolution down to 10 nm in the lateral direction, and 30–50 nm in the axial direction. However, there is one important requirement to perform this type of imaging: making dye molecules blink. This usually relies on the utilization of complex buffers, containing different chemicals and sensitive enzymatic systems, limiting the reproducibility of the method. We report here that the commercial mounting medium Vectashield can be used for STORM of Alexa-647, and yields images comparable or superior to those obtained with more complex buffers, especially for 3D imaging. We expect that this advance will promote the versatile utilization of 3D STORM by removing one of its entry barriers, as well as provide a more reproducible way to compare optical setups and data processing algorithms.

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