Spatial light modulators have become an essential tool for advanced microscopy, enabling breakthroughs in 3D, phase, and super-resolution imaging. However, continuous spatial-light modulation that is capable of capturing sub-millisecond microscopic motion without diffraction artifacts and polarization dependence is challenging. Here we present a photothermal spatial light modulator (PT-SLM) enabling fast phase imaging for nanoscopic 3D reconstruction. The PT-SLM can generate a step-like wavefront change, free of diffraction artifacts, with a high transmittance and a modulation efficiency independent of light polarization. We achieve a phase-shift > π and a response time as short as 70 µs with a theoretical limit in the sub microsecond range. We used the PT-SLM to perform quantitative phase imaging of sub-diffractional species to decipher the 3D nanoscopic displacement of microtubules and study the trajectory of a diffusive microtubule-associated protein, providing insights into the mechanism of protein navigation through a complex microtubule network.

Institute of Biotechnology of the Czech Academy of Sciences BIOCEV Vestec Prague West 25250 Czech Republic

Institute of Photonics and Electronics of the Czech Academy of Sciences Prague 18251 Czech Republic

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Surpassing the Diffraction Limit in Label-Free Optical Microscopy

Electro-detachment of kinesin motor domain from microtubule in silico

Fast photothermal spatial light modulation for quantitative phase imaging at the nanoscale

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figshare
10.6084/m9.figshare.14170622