A diffraction-limited scanning system providing broad spectral range for laser scanning microscopy

Abstract

Diversified research interests in scanning laser microscopy nowadays require broadband capability of the optical system. Although an all-mirror-based optical design with a suitable metallic coating is appropriate for broad-spectrum applications from ultraviolet to terahertz, most researchers prefer lens-based scanning systems despite the drawbacks of a limited spectral range, ghost reflection, and chromatic aberration. One of the main concerns is that the geometrical aberration induced by off-axis incidence on spherical mirrors significantly deteriorates image resolution. Here, we demonstrate a novel geometrical design of a spherical-mirror-based scanning system in which off-axis aberrations, both astigmatism and coma, are compensated to reach diffraction-limited performance. We have numerically simulated and experimentally verified that this scanning system meets the Marechal condition and provides high Strehl ratio within a 3 degrees x 3 degrees scanning area. Moreover, we demonstrate second-harmonic-generation imaging from starch with our new design. A greatly improved resolution compared to the conventional mirror-based system is confirmed. This scanning system will be ideal for high-resolution linear/nonlinear laser scanning microscopy, ophthalmoscopic applications, and precision fabrications.

Multiphoton imaging to identify grana, stroma thylakoid, and starch inside an intact leaf

“…The experimental setup is shown in Figure 5, which is similar to our previous reports [23,38]. This setup allows the simultaneous measurement of SHG and 2PF in the forward and backward directions.…”