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2 Janelia Publications

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    Ji Lab
    04/15/18 | Three-photon fluorescence microscopy with an axially elongated Bessel focus.
    Rodriguez C, Liang Y, Lu R, Ji N
    Optics Letters. 2018 Apr 15;43(8):1914-1917. doi: 10.1364/OL.43.001914

    Volumetric imaging tools that are simple to adopt, flexible, and robust are in high demand in the field of neuroscience, where the ability to image neurons and their networks with high spatiotemporal resolution is essential. Using an axially elongated focus approximating a Bessel beam, in combination with two-photon fluorescence microscopy, has proven successful at such an endeavor. Here, we demonstrate three-photon fluorescence imaging with an axially extended Bessel focus. We use an axicon-based module that allowed for the generation of Bessel foci of varying numerical apertures and axial lengths, and apply this volumetric imaging tool to image mouse brain slices and for in vivo imaging of the mouse brain.

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    Ji Lab
    04/01/18 | 50 Hz volumetric functional imaging with continuously adjustable depth of focus.
    Lu R, Tanimoto M, Koyama M, Na J
    Biomedical Optics Express. 2018 Apr;9(4):1964-76. doi: 10.1364/BOE.9.001964

    Understanding how neural circuits control behavior requires monitoring a large population of neurons with high spatial resolution and volume rate. Here we report an axicon-based Bessel beam module with continuously adjustable depth of focus (CADoF), that turns frame rate into volume rate by extending the excitation focus in the axial direction while maintaining high lateral resolutions. Cost-effective and compact, this CADoF Bessel module can be easily integrated into existing two-photon fluorescence microscopes. Simply translating one of the relay lenses along its optical axis enabled continuous adjustment of the axial length of the Bessel focus. We used this module to simultaneously monitor activity of spinal projection neurons extending over 60 µm depth in larval zebrafish at 50 Hz volume rate with adjustable axial extent of the imaged volume.

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