Dual Independent Enhanced Scan Engines for Large field-of-view Two-Photon imaging (Diesel2p)

Type: Optics / Microscopy,

Keywords: Two-Photon, Multiphoton, Microscopy, Microscope, Mouse, Neuroimaging, Subcellular, Open-Source, Mesoscope, Brain

Diesel2p is a large field-of-view two-photon microscope capable of subcellular resolution, with dual independent scan engines for highly flexible, asymmetric multi-point sampling of distributed neural circuitry.

Diesel2p is a custom two-photon microscope system with dual scan engines that can operate completely independently. Each arm has optical access to the same large imaging volume (~25 mm^2 FOV) over which subcellular resolution is maintained in scattering tissue to typical 2-photon imaging depths. Each arm can use multiple sources simultaneously, for example, in imaging and photoactivation experiments.

*Two-Photon system with a ~25 mm^2 FOV to encompass multiple cortical areas and provides subcellular resolution throughout.
*Simultaneous two-region imaging using two scan engine arms, with each arm being completely independent and reconfigurable.
*Equipped with adaptive optics to compensate aberration caused by tissue scattering and to enable rapid volumetric imaging.
*Dual excitation light sources for dual-color imaging or simultaneous all-optic imaging and optogenetics.
*8-mm working distance air-immersion objective facilitates animal imaging and compatibility with other probe instruments.

Diesel2p is a newly developed imaging system to enable flexible, simultaneous, multi-region, multiphoton excitation in scattering tissue. The imaging pathways allow for neural recording in two selected portions of the imaging volume simultaneously. This new system is also designed to facilitate experiments with behaving animals. The objective lens is air immersion, so no water interface is required, and it has an 8-mm-long working distance, to enable a variety of headplate designs and other instrumentation that needs to be close to the imaged area (e.g., electrode arrays).

*In vivo large field neural imaging of neurons and neurites.
*Simultaneous two distant region imaging with independent imaging parameters.
*Simultaneous all-optic imaging and optogenetics.


*Large field-of-view of ~25 mm^2
*Dual independent scan engines
*Adaptive optics for wavefront shaping and rapid volumetric imaging
*Capable of subcellular resolution
*Achromatic for dual excitation light sources
*Open source

While suitable for imaging in small mammals, two-photon imaging depth limits some experiments in larger mammals.

The system requires a laser capable of two-photon excitation, as well the manufacture of custom open-source optics.

Yu et al. 2021, Diesel2p mesoscope with dual independent scan engines for flexible capture of dynamics in distributed neural circuitry. Nature Communications. doi: https://doi.org/10.1038/s41467-021-26736-4


Spencer Smith, Associate Professor


University of California, Santa Barbara, Santa Barbara, CA



Che-Hang Yu, Postdoc, University of California, Santa Barbara
Jeffrey N. Stirman, Director of Imaging, LifeCanvas Technologies
Yiyi Yu, Postdoc, University of California, Santa Barbara
Riichiro Hira, Postdoc, University of California, Santa Barbara
Spencer L. Smith, Associate Professor, University of California, Santa Barbara



McKnight Foundation Human Frontier Science Program (RGP0027/2016)
NSF (NeuroNex #1934288 and BRAIN EAGER #1450824)
NIH (R01NS091335 and R01EY024294)