Brain slices were imaged via two-photon excitation microscopy using an Olympus BX51 microscope with a 60× (1.0 NA) objective. Infrared excitation light was provided via a Ti:sapphire laser system (Mai Tai HP1040; Spectra Physics), and non-de-scanned fluorescence emission was acquired via dual photomultiplier tubes. The microscope was equipped with a dual scanhead; one x–y galvanometer mirror system controlled the imaging laser beam while another x–y galvanometer system independently positioned a photostimulation (405 nm; OBIS FP LX; Coherent) beam for photolysis of PA-Nic (Banala et al., 2018 (link)). For two-photon experiments, all aspects of imaging, photostimulation, and recording were conducted using Prairie View 5.4 software (Bruker Nano).
Mai tai hp1040
Manufactured by Spectra-Physics
The Mai Tai HP1040 is a tunable, ultrafast Ti:Sapphire laser from Spectra-Physics. It provides 1 watt of average output power and a pulse width of less than 40 femtoseconds across a tunable wavelength range of 690 to 1040 nanometers.
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2 protocols using mai tai hp1040
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Two-Photon Imaging and Photostimulation
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Two-Photon Imaging and Photostimulation
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Brain slices were imaged via two-photon excitation microscopy using an Olympus BX51 microscope with a 60× (1.0 NA) objective. Infrared excitation light was provided via a Ti:sapphire laser system (Mai Tai HP1040; Spectra Physics), and non-de-scanned fluorescence emission was acquired via dual photomultiplier tubes. The microscope was equipped with a dual scanhead; one x–y galvanometer mirror system controlled the imaging laser beam while another x–y galvanometer system independently positioned a photostimulation (405 nm; OBIS FP LX; Coherent) beam for photolysis of PA-Nic (Banala et al., 2018 (link)). For two-photon experiments, all aspects of imaging, photostimulation, and recording were conducted using Prairie View 5.4 software (Bruker Nano).
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