Two-photon in vivo physiology

About the infrastructure

Two-photon in vivo physiology offers a combination of multiphoton and imaging systems at high spatio-temporal resolution, combined with other physiological equipment for electrophysiology and imaging, in the living organism (mammals, zebrafish, etc), but solutions also for in vitro experiments can be discussed.

This research platform is currently unique in Scandinavia in providing researchers access to the most advanced imaging tools for study and perturb in a controlled way cellular dynamics at high spatio-temporal resolution in vivo but also in vitro (e.g. organoids), combined with technical assistance and indication about data analysis options. With its superior penetration possibility, reduced photo damage and out of focus bleaching, and with the high speed, our multiphoton systems can be used for high resolution quantitative image analysis of biological processes at cellular and subcellular resolution in different organs and physiological and pathological conditions.

Two-photon in vivo physiology has the potential to boost biomedical research in neuroscience (also in behaving animals), retinal and ophthalmologic research, endocrine physiology, tumour and cell biology in investigating and analysing their sample preparations with a spatial and temporal resolution not attainable by standard optical equipment. Research can be combined with electrophysiology and high resolution histology. Two-photon imaging can be applied in the living organism by the use of chronic optical windows, but also used in vitro (organoids, cell cultures etc.).

The research infrastructure is located in the open area in the UCCB building, with possibility to store animals with chronic windows for longitudinal imaging in a dedicated room within the same building so to move them to the imaging/physiology labs easily in multiple sessions for chronic/longitudinal imaging.

Equipment and services

• A conventional two photon system (Scientifica, UK, laer: 700-1000 nm, green and red PMTs); and two high speed two-photon imaging systems (resonant scanning, with 700-1300 nm laser, with green/red/blue  PMTs; and a 3D-acousto-optic modulator system, both from Femtonics; Hungery) for functional calcium imaging.
• For awake behaving studies we have a virtual reality system that can be connected with the two-photon microscope as well a fiberscopic imaging and photostimulation system for optogenetics in freely moving animals (Inscopix nVoke).
• In one system (Femtonics Resonant), a 3D two-photon holographic module for photostimulation of functionally identified cells (e.g. by calcium imaging) is also installed.
• Intra and extracellular electrophysiology is combined in our multiphoton set ups.
• Camera-based imaging (intrinsic signal and voltage-sensitive dye) that can be used to target virus injections in a BSL-2 hood to express calcium indicators and opsins in a stereotaxic apparatus for brain injections but also in other organs.
• Dedicated set-up for retinal physiology (electroretinography, pattern and flash) that is combined with a retinal imaging microscope. The system includes a transpupillary laser
• Dedicated objectives for imaging of clarified tissues, as well as 3D reconstruction software.
• Research support in form av consultation for data analysis.

Access 

Access to two-photon in vivo physiology is limited, please contact Anushree Tripathi or Paolo Medini to learn more and get access to the infrastructure.