We have designed and built a time-gated continuous wave stimulated emission depletion (CW-STED) nanoscopy to visualize microstructures beyond the diffraction limit. An off-line time-gating detection was performed with the help of time-correlated single-photon counting technique. Experimental results showed that before time-gating, the resolution of our system was about 75 nm with a depletion beam (592 nm) power of 200 mW. By using the off-line time-gating detection, the resolution was further improved to 38 nm. Biological samples were also used to test the performance of our time-gated CW-STED, and a resolution of 70 nm was achieved with a depletion beam (592 nm) power of 85 mW. Detailed principles of time-gated CW-STED were discussed in the text. The time-gated STED provides a better resolution with finite laser power.
Stimulated emission depletion microscopy (STED) has been proved to be a feasible and straightforward method of breaking the conventional diffraction barrier in the far-field. In this paper, we design and setup a home-built high speed STED microscope. Both the excitation beam (488nm) and the depletion beam (592nm) are provided by continuous wave lasers. By using a pair of galvo mirrors (TILL Yanus IV Digital Scan Head), we realize a pixel dwell time down to 2 μs in the experiment which enables an acquisition speed of 2 frames per second in an imaging field of 5*5 μm with an individual pixel size of 10nm. The image acquisition process is controlled by the software Imspector. In the manuscript, we give a clear description on how to build the microscope and also conduct several experiments to evaluate its performance in practice. A spatial resolution of <100nm, which is well beyond the diffraction barrier has been demonstrated in both nanoparticles and biomedical samples. Featuring a superresolution together with a high imaging speed, our STED microscope has big potential to be widely applied in related scientific researches.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.