Mesoscale imaging (MVX-10 epifluorescence microscope; Olympus, Tokyo Japan) was performed to visualize stimulation-induced activity on a large spatial scale (3.7 × 3 mm) [24 (link), 29 , 36 (link)]. Time series images were captured at 20 frames per second (fps) using a charge-coupled device (CCD) camera (Retiga, R1 18 imaging). A two-photon microscope (Bruker, Madison, WI) equipped with an optical parametric oscillator (OPO) laser (Insight DS+, Spectra Physics, Menlo Park, CA) at 920 nm and a 16 × 0.8 numerical aperture (NA) water immersion objective (Nikon, Melville, NY) was employed to answer questions related to the different temporal calcium dynamics with cellular resolution (407 × 407 μm, 0.8 μm pixel−1; 30 fps).
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Mesoscale and Two-Photon Imaging Techniques
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Other organizations : University of Pittsburgh, Center for the Neural Basis of Cognition, Carnegie Mellon University, University of Wisconsin–Madison, Neurological Surgery, Wisconsin Institutes for Discovery, McGowan Institute for Regenerative Medicine
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Variable analysis
independent variables
- Stimulation-induced activity
- Stimulation-induced activity visualized on a large spatial scale (3.7 × 3 mm)
- Temporal calcium dynamics with cellular resolution (407 × 407 μm, 0.8 μm pixel^-1; 30 fps)
- Charge-coupled device (CCD) camera (Retiga, R1 18 imaging)
- Two-photon microscope (Bruker, Madison, WI) equipped with an optical parametric oscillator (OPO) laser (Insight DS+, Spectra Physics, Menlo Park, CA) at 920 nm and a 16 × 0.8 numerical aperture (NA) water immersion objective (Nikon, Melville, NY)
- Positive control: Not specified
- Negative control: Not specified
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