Invigilo™-3 Wide-Field Imaging System

Introducing Invigilo-3: improved illumination, enhanced software functions

Invigilo-3 is the only commercial solution optimized for wide-field imaging in awake-behaving mice.  The system facilitates simultaneous multi-channel imaging and behavioral observation by integrating optical components with locomotion tracking and automated stimuli delivery.

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      Invigilo-3 offers a cortex-wide window into the brain of an awake and behaving mouse.

      The system is tailored for neuroscientists seeking an efficient, adaptable, and cost-effective imaging solution.

      Imaging of neuronal or glial calcium activity alongside hemodynamics with Invigilo™-3

      Recent advances in fluorescent probes and craniotomy preparations have enhanced wide-field imaging capabilities. As a result, we can now perform minimally invasive recordings of calcium signals across the entire cortex with a significantly higher sampling rate than that of the laser-scanning techniques. Some of the options for the new cranial preparations include the “crystal skull” described by Kim et al., 2016, “transparent polymer skull” by Ghanbari et al., 2019 or “transparent skull” by Vesuna S. et al., 2020. All of these can be implemented in the Invigilo™-3.

      Moreover, Invigilo-3 captures optical BOLD (blood oxygenation level-dependent) signal simultaneously with fluorescence imaging in multiple cortical areas. Besides providing a functional readout for blood flow, the BOLD signal enables the correction of fluorescence signal distortions (e.g., GCaMP6-based Ca2+ signal) due to activity-dependent light adsorption by hemoglobin. For more on the need for BOLD correction see West S.L. et al., 2022 or Valley M.T. et al., 2020.

      BOLD-corrected calcium signal - wide field imaging in awake mice

      Integrating functional imaging and behavioral observation

      Wide-field imaging integrated with behavior in the Invigilo system

      Optimized for awake, head-fixed mice, Invigilo-3 facilitates the integration of wide-field imaging and behavioral experiments as part of a holistic neuroscience approach. Its robust frame and head fixation combined with an air-lifted flat-floored cage, enable two-dimensional locomotion, reduce stress, and provide maximal mechanical stability.

      Behavioral monitoring relies on a magnetic high-speed (200 fps), high spatial resolution (<0,5 mm) locomotion-tracking device built into the air dispenser. Tracking works both in the dark and in light conditions. What is more, locomotion tracking facilitates other behavioral readouts (open field, object recognition, social interactions, maze navigation). The tracking software supports automated stimulus delivery and enables closed-loop experiments.

      What gives us confidence that the Inviglo™-3 is optimal for integrating imaging and behavioral research? – The system was developed by the same team that developed the Mobile HomeCage, which is used by 220+ labs worldwide. Moreover, Neurotar’s contract research team uses Invigilo™-3 for service provision to the pharmaceutical industry. CRO team’s feedback was invaluable for the development of the system.

      Invigilo-3 features an LED ring around the objective for even illumination, ideal for longitudinal brain imaging with transparent skull preparations. The ring was developed in response to the challenges of ensuring even illumination experienced by our and other academic research teams (see e.g., Doran et al., 2023).

      Software integration for data acquisition and automated functional connectivity analysis

      Invigilo’s software streamlines the workflow by synchronizing all hardware components through a user-friendly interface. It simplifies managing settings for sCMOS cameras and LED light sources. The software’s new advanced analysis functions include automatic correction for hemodynamics,  ΔF/F measurements of changes in fluorescence intensity, and functional connectivity analysis. All of these make it easier to draw meaningful conclusions about the brain’s functional connectivity from complex datasets.

      The images above illustrate the steps in the functional connectivity analysis, starting with the segmentation of data into active locomotion vs rest (top left), calcium-activity-based cluster mapping (top middle), correlation of locomotor and calcium activity (bottom left), and changes in the calcium activity correlation between the clusters (bottom middle and right).

      Invigilo™-3 highlights

      • Multi-channel imaging: Includes an epifluorescent channel (red or green), two backscatter channels (red and green), and LSCI for specific configurations.
      • Effortless focusing: The integrated motorized vertical breadboard mounts the optical assembly, including the objective, LEDs, and camera, simplifying the focusing process.
      • LED ring for even illumination: The Invigilo-3 features an LED ring around the objective, ensuring even and reproducible illumination for backscatter imaging.
      • Stable recordings: A robust aluminum frame paired with an adjustable head-fixation system ensures mechanical stability and reduces stress on the mouse.
      • Innovative cage design: The air-lifted cage with a flat floor encourages natural exploratory behavior, minimizing stress and enhancing the quality of imaging.
      • Near-panoramic view: A 270-degree unobstructed view allows for versatile applications, such as visual stimulation, virtual reality, behavior video recording, or pupil dilation monitoring.
      • Advanced software capabilities: New functions for automated analysis of the brain’s functional connectivity enhance research efficiency and accuracy.

      What Invigilo™ stands for?

      Invigilo is related to vigilant; both are derived from the Lat. verb vigilare, which means ‘to stay/keep awake’. 

      Is Invigilo™-3 good for you?

      Want to start imaging in vivo on a budget?

      At a fraction of the price of a two-photon microscope, Invigilo-3 is a cost-effective solution for you.

      Want to supplement two-photon imaging data with a dynamic picture of the entire cortex?

      Invigilo-3 is complementary because it offers a broader, cortex-wide field of view.

      Want to augment functional and morphological readouts with behavioral observation?

      Invigilo-3 is a way to start with a ready-to-go solution integrating multi-channel imaging and behavior in awake head-fixed mice.

      Want to learn more or get a quote? – Contact us to schedule an online meeting.