Ultrafast photon counting applied to resonant scanning STED microscopy |
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| Authors: | XUNDONG WU LIGIA TORO ENRICO STEFANI YONG WU |
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| Affiliation: | 1. Division of Molecular Medicine, Department of Anesthesiology, David Geffen School of Medicine, University of California, Los Angeles, California, U.S.A.;2. Department of Molecular & Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, California, U.S.A.;3. Cardiovascular Research Laboratory, David Geffen School of Medicine, University of California, Los Angeles, California, U.S.A.;4. Department of Physiology, David Geffen School of Medicine, University of California, Los Angeles, California, U.S.A. |
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| Abstract: | To take full advantage of fast resonant scanning in super‐resolution stimulated emission depletion (STED) microscopy, we have developed an ultrafast photon counting system based on a multigiga sample per second analogue‐to‐digital conversion chip that delivers an unprecedented 450 MHz pixel clock (2.2 ns pixel dwell time in each scan). The system achieves a large field of view (～50 × 50 μm) with fast scanning that reduces photobleaching, and advances the time‐gated continuous wave STED technology to the usage of resonant scanning with hardware‐based time‐gating. The assembled system provides superb signal‐to‐noise ratio and highly linear quantification of light that result in superior image quality. Also, the system design allows great flexibility in processing photon signals to further improve the dynamic range. In conclusion, we have constructed a frontier photon counting image acquisition system with ultrafast readout rate, excellent counting linearity, and with the capacity of realizing resonant‐scanning continuous wave STED microscopy with online time‐gated detection. |
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| Keywords: | Fluorescence microscopy photon counting resonant scanning STED super‐resolution |
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