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3D printed high-NA ultra-thin catadioptric condenser to minimize background contribution from cross-phase modulation in stimulated Raman scattering microscopy (Conference Presentation)
Author(s): Andrea Bertoncini; Carlo Liberale
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Paper Abstract

To avoid the unwanted non-resonant background contribution induced by cross-phase modulation (XPM) in Stimulated Raman Scattering (SRS) microscopy [1], the collection of forward transmitted signal is normally done with a microscope objective having a numerical aperture (NA) higher than the objective used for excitation. However, while high NA microscope objectives are usually bulky and expensive, because a complex design is needed to achieve good optical imaging performances, only a capability to collimate highly divergent beams is needed for forward detection of signal in SRS microscopy. Additionally, because high NA microscope objectives have a short working distance and are bulky, their use as forward collecting optical element is not compatible with tightly closed top-stage incubators, as used in live-cell experiments. Here we show the use of a high NA 3D printed ultra-thin optical lens, composed of micro-reflective and -refractive elements, to replace commercial high NA microscope objectives for forward collection of signal in Stimulated Raman Scattering microscopy. The lens is fabricated on a 170µm thick coverslip with direct laser writing based on Two-Photon Lithography with a commercial system (Nanoscribe) and using the proprietary IP-S photoresist. It has a thickness of 300 µm and a diameter of 1 cm. Thanks to its compactness, this optical element can easily fit inside top-stage microscope incubators. The resulting NA of this catadioptric condenser lens is 1.2 when working in water immersion. We show the complete removal of the non-resonant XPM contribution from SRS spectra of incubated cells. [1] Ji-Xin Cheng, Xiaoliang Sunney Xie, Coherent Raman scattering microscopy, CRC press, 2016.

Paper Details

Date Published: 4 March 2019
Proc. SPIE 10882, Multiphoton Microscopy in the Biomedical Sciences XIX, 1088229 (4 March 2019); doi: 10.1117/12.2510085
Show Author Affiliations
Andrea Bertoncini, King Abdullah Univ. of Science and Technology (Italy)
Carlo Liberale, King Abdullah Univ. of Science and Technology (Saudi Arabia)

Published in SPIE Proceedings Vol. 10882:
Multiphoton Microscopy in the Biomedical Sciences XIX
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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