3D printed high-NA ultra-thin catadioptric condenser to minimize background contribution from cross-phase modulation in stimulated Raman scattering microscopy (Conference Presentation)

Andrea Bertoncini; Carlo Liberale

doi:10.1117/12.2510085

4 March 2019 3D printed high-NA ultra-thin catadioptric condenser to minimize background contribution from cross-phase modulation in stimulated Raman scattering microscopy (Conference Presentation)

Andrea Bertoncini, Carlo Liberale

Author Affiliations +

Proceedings Volume 10882, Multiphoton Microscopy in the Biomedical Sciences XIX; 1088229 (2019) https://doi.org/10.1117/12.2510085
Event: SPIE BiOS, 2019, San Francisco, California, United States

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.

Conference Presentation

Citation Download Citation

Andrea Bertoncini and Carlo Liberale "3D printed high-NA ultra-thin catadioptric condenser to minimize background contribution from cross-phase modulation in stimulated Raman scattering microscopy (Conference Presentation)", Proc. SPIE 10882, Multiphoton Microscopy in the Biomedical Sciences XIX, 1088229 (4 March 2019); https://doi.org/10.1117/12.2510085

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KEYWORDS

Microscopy

Raman scattering

Combined lens-mirror systems

Microscopes

Modulation

Objectives

Optical components

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