Automatic tube lens design from stock optics for microscope remote-refocusing systems

Abstract

The remote-refocusing approach of Botcherby et al. [Opt. Lett. 32, 2007 (2007)10.1364/OL.32.002007] has been applied widely to 2D and 3D fluorescence microscopes to enable rapid refocusing of the optical system without mechanically perturbing the sample. In order for this approach to operate correctly, it requires that the overall magnification of the first two microscope systems matches the ratio of the refractive indices in sample and intermedia image spaces. However, commercially available tube lenses are not always suitable to produce the desired overall magnification. Therefore, a practical approach to produce tube lenses with low expense and diffraction-limited performance is required. Tube lenses can be formed using a pair of stock achromatic doublets, however, selecting appropriate pairs of achromatic doublets from stock optics is a time-consuming process, as many combinations can be considered. In this paper, we present two software packages (Catalogue Generator and Doublet Selector) developed in MATLAB that use the application programming interface (ZOS-API) to the Zemax OpticStudio optical design software to realise an automatic search of stock achromatic doublets to produce microscope tube lenses with a specified focal length, entrance pupil diameter and maximum design field angle. An algorithm to optimise principal plane positions in versions of OpticStudio before 20.2 was also introduced to enable the use of older software versions. To evaluate the performance of Catalogue Generator and Doublet Selector, we used them to generate ten tube lens designs. All of the software-produced tube lenses have a better optical performance than those using manually selected pairs of stock doublets lenses.

Axial de-scanning using remote focusing in the detection arm of light-sheet microscopy

“…Following the footsteps of Hong et al 45 we considered how our off-axis remote-focusing design might affect the collection efficiency 46,47 and therefore the achievable DL range (Supplementary Fig.10). As it turns out, because in our current design the two images after the Obj2 are formed at the far end of the FOV, some of the rays always miss the Obj2 when we try to defocus.…”

“…Following the footsteps of Hong et al 45 we considered how our off-axis remote-focusing design might affect the collection efficiency 46,47 and therefore the achievable DL range (Supplementary Fig. 10).…”

Signal Improved ultra-Fast Light-sheet Microscope (SIFT) for large tissue imaging

“…The laser beam then passes through a pair of relay lenses and splits into two identical remote focusing [48][49][50] arms through a second PBS. The two relay lenses, 200 mm (ACT508-200-A-ML, ThorLabs) and 75 mm (AC508-075-A-ML) forms a 4f system and conjugates the pupil of objective 1 to the pupil of the remote focusing objective (Olympus UMP PlanFl 10x, NA 0.30) 51 . Each remote focusing arm consists of a QWP, an objective and a mirror.…”

Signal improved ultra-fast light-sheet microscope for large tissue imaging

“…The laser beam then passes through a pair of relay lenses and splits into two identical remote-focusing [59][60][61] arms through a second PBS. The two relay lenses, 200 mm (ACT508-200-A-ML, ThorLabs) and 75 mm (AC508-075-A-ML) form a 4 f system and conjugate the pupil of objective 1 to the pupil of the remote-focusing objective (Olympus UMP PlanFl 10×, NA 0.30) 62 . Each remote-focusing arm consists of a QWP, an objective and a mirror.…”