OSA Optical Design and Fabrication Congress 2021 ©

IM3A.5.pdf
OSA 2021

Laser Scanning Microscope with Large Field and High NA.

David Kessler
Kessler Optics & Photonics Solutions, Ltd.
Dave.Kessler@KesslerOptics.com

Abstract: The design is telecentric, all reflective, highly symmetrical, with no off-axis aberrations
and has a large working distance. It uses a free form element at the input and a highly efficient
Offner-like collector. © 2021 David Kessler

1. Summary
1.1. Background
There is a need [1,2] for a large field, efficient and low noise system for Two Photon Microscopy (TPM) especially
in the area of brain research. There is also an advantage for a reflective objective to minimize pulse broadening [3].
The Apparatus described herein provides for a 30 mm field or more, keeping the NA at or above 0.5. The design is
highly symmetrical about the scanning mirror and thus there are no off-axis aberrations and no field changes due to
objective dispersion. The system is essentially on-axis for all scanning angles. The on-axis aberrations are corrected
by a free-form pre-scan compensator placed on the input beam. An arcuate scan line is produced at the sample plane,
concentric with the axis of symmetry of the scan mirror. The whole system is linearly translated in the cross-scan
direction to capture the full two-dimensional field.
1.2 The scanning system
Fig. 1 shows the toroidal mirror and the axis of symmetry on which the scanning mirror is placed. The scan mirror is
shown at three scan angles. Since the toroidal mirror power is larger in the cross-scan direction, the beam coming
from bellow towards the scan mirror, needs to be anamorphic, namely, with a smaller dimension along the axis of
symmetry. The toroidal is totally symmetrical about the axis shown and has a number of aspherical coefficients in
the cross-scan direction. This toroidal can also be regarded as a segment of an even-aspheric surface with its vertex
on the axis of symmetry. The scanning beam is image-telecentric. At the image, a rectangular cranial cover glass is
shown covering a depth of 300 microns of water, representing the brain.

Fig. 1. The scanning system showing the scan mirror and the toroidal primary mirror

1.3 The input beam pre-compensator

The toroidal mirror introduces aberrations which are pre-compensated prior to the scan mirror as shown on Fig 2.
The compensator shown in this example has two refractive elements: a cylinder lens to generate the anamorphic
beam shape at the scan mirror, and a free-form element which is a polynomial element with asymmetrical
coefficients in the cross-scan direction. The refractive compensator can be replaced with a reflective one, so the
whole system is reflective and thus with minimal pulse broadening due to dispersion. The pre-compensator can also
include a dynamic Adaptive Optics (AO) element to allow exploration of different depths at the image as is done
for example in [4].
1.4 Collection with an Offner-like collector.
The common collection method in laser scanning is to collect the emitted light by retroreflecting towards the input
and split it off with a dichroic splitter onto a small low noise detector. This can be used also with the system above.
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Fig. 2. The pre-compensator shown with the scan mirror and the toroidal primary.

Another option is to take advantage of the arcuate scan line and recognize that it can be imaged with high
collection efficiency into a thin arcuate detector using an Offner-like system [5] (the arcuate detector can be
implemented with a small round detector and a round to line converter such as a fiber-based one) . Fig. 3 shows the
scanning microscope with the Offner-like collector. The input beam (a) shown in red is modified by the
compensator (b), reflected off the folding mirror (c) and the scan mirror (d). Fig. 4 is a cross section of the single
element comprising the objective and the collector. This is an even-aspheric element where its bottom surface (e) is
dichroically coated to reflect the IR scanning beam onto the 30-mm-long arcuate scan line at the sample (f). This
bottom surface (e) transmits the emitted, shorter wavelength light, shown in blue, towards the Offner concave
primary mirror (g), then onto the convex Offner secondary (h) and again onto the primary (i) and towards a thin
arcuate detector (j). Multiple scan lines can be simultaneously imaged by the Offner system onto multiple arcuate
detectors.

Fig. 3. The full system with the collector. Fig. 4. The objective/collector element

2. Acknowledgements:
This work began thanks to suggestions and insightful input from Dr. Justin Little at NYU Langone Health.

3. References
[1] Jonathan R. Bumsted et. al. “Designing a large field of view two photon microscope using optical invariant,” Neurophotonics 5(2)
025001 (Apr-Jun 2018).
[2] Robert S. Tsai et al. “Ultra-large field -of-view two photon microscopy,” Optics Express Vol. 23, No.11 Jun 2015).
[3] J. Diels and W. Rudolf, Ultrashort Laser Pulse Phenomena, 2nd ed. (Academic Press, Cambridge, MA 2006).
[4] Congping Chen, et. al., “Adaptive optics two-photon microscopy for in vivo imaging of cortex and
hippocampus in mouse brain,” Proc. SPIE Vol. 10865,108650M (1 March 2019)
[5] Abe Offner, “Unit power Imaging Catoptric Anastigmat,” USA Patent 3,748,015, July 1973.