Friday 8th of July - 16:10
nTCscope

Recently, we presented a non-telecentric (nTC) ultra-low-cost solution for 3D random access mesoscale imaging in two-photon microscopy [1]. Using a standard x20 objective, this nTC- approach allowed increasing the field of view from typically 0.5 mm to 3.5 mm, with a corresponding elongation of the point spread function (PSF). With the addition of an electrically tunable lens, we further achieved a z-range of 600 μm within 2 ms, all without strongly distorting the excitation volume or efficiency. While this approach has already been useful in a range of experimental works [1-5], the issue remained that the PSF would grow with the expansion of the field of view, thus limiting spatial resolution.

Here, we present a next step in the optimisation of nTC: expansion of the field of view without affecting PSF size. Still using the same x20 objective, we maintain a diffraction limited resolution up to a field of view of ~0.8 mm. This for example allows us to image the entire length of a larval zebrafish brain while comfortably maintaining single cell resolution.

[1] FK Janiak, P Bartel, MR Bale, et al. Non-telecentric two-photon microscopy for 3D random access mesoscale imaging. Nat Commun 13, 544 (2022)

[2] T Yoshimatsu, P Bartel, C Schröder, FK Janiak, F St-Pierre, P Berens, T Baden, “Ancestral circuits for vertebrate color vision emerge at the first retinal synapse”, Science Advances Vol 7, Issue 42 (2021)

[3] M Zhou, J Bear, PA Roberts, FK Janiak, J Semmelhack, T Yoshimatsu, T Baden, “Zebrafish Retinal Ganglion Cells Asymmetrically Encode Spectral and Temporal Information across Visual Space”, Current Biology 30, issue 15, P2927- 2942.E7, August 03, 2020

[4] P Bartel, FK.Janiak, D Osorio and T Baden, “Colourfulness as a possible measure of object proximity in the larval zebrafish brain” Current Biology 31(5), R235-R236, March 8, 2021

[5] P Bartel, T Yoshimatsu, FK Janiak, T Baden, Spectral inference reveals principal cone-integration rules of the zebrafish inner retina (2021), Current Biology 31(23), 5214-5226,

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