Two-photon calcium imaging was conducted via a Bergamo II multi-photon microscope (THORLABS). Ti:Saphire pulsed laser (Coherent) with wavelength of 920 nm and power of ~80 mW (measured at the tissue) was used to excite the calcium indicators. Scaning of the field of view was done by Galvo-Resonant X-Y mirrors. A ×16 water-immersion objective lens (Nikon) with numerical aperture of 0.8 was used for imaging. The emitted light from calcium indicators was collected via a GaAsP photomultiplier tube (Hamamatsu). The field of view size was 835×835 µm, and frames were captured at spatial resolution of 800×800 pixels and temporal resolution of 19.6 Hz. The depths of imaging were aimed between 110 and 190 µm (layers II/III).
Bergamo 2 multi photon microscope
Manufactured by Thorlabs
The Bergamo II multi-photon microscope is a high-performance imaging system designed for advanced fluorescence microscopy applications. The core function of this microscope is to enable multi-photon excitation and imaging of samples, allowing for deep tissue penetration and reduced photodamage compared to traditional single-photon microscopy techniques.
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Lab products found in correlation
2 protocols using bergamo 2 multi photon microscope
1
Two-photon calcium imaging of neuronal activity
2
Multiphoton Imaging of Superficial RSC Neurons
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Population of RSC neurons at 100-200 mm from dorsal surface (for superficial agranular RSC) were imaged using a Bergamo II multiphoton microscope (Thorlabs). A Ti:Sapphire excitation laser (Coherent) was operated at 920 nm ($20-120 mW laser power at the sample) through a 16x lens (NA = 0.8, Nikon). Green fluorescence from GCaMP6s was collected and measured with a GaAsP photomultiplier tubes (PMTs). Blackout fabric was used to prevent stray light from entering the objective and PMTs. Images were collected at $20 frames per second. Imaging window was centered at around 2.5 mm from the traverse sinus and 0.5 mm from the midline sinus. The field of view was 835 3 835 mm in size.
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