Speakers
Description
Multi photon scanning microscopy gives access to high axial resolution, high penetration depth and
the third spatial dimension at the cost of imaging speed as each pixel must be probed individually.
The image acquisition time of conventional scanning systems is inertia limited by the oscillation
frequency of two galvanometric mirrors, making observation of biological processes that occur in
three-dimensional space on a milli second timescale inaccessible.
We present the prototype in development of a mobile, small footprint Spectro-Temporal Laser
Imaging by Diffractive Excitation (SLIDE) system that aims to overcome this limitation. This multi-
photon scanning microscope utilizes a fast tunable Fourier Domain Mode Locking (FDML) Laser with
a center wavelength of 1065 nm and a pulse repetition rate up to 1 GHz in combination with a
diffractive element to achieve frame rates of up to 1.8 kHz and live volume imaging of a 250
x 250 x 100 μm³ volume with a volume rate of up to 20 volumes per second.
At these scanning speeds, corresponding to a pixel dwell time of ~1.2 ns each pixel only gets excited
a single time per acquisition, limiting the photon budget. We compare this Imaging modality to a
conventional multi photon system and demonstrate its feasibility for in vitro and in vivo imaging.
