Figure 1. Specifications for the 2p-RAM.
(A) Schematic of the dorsal surface of the mouse cortex (from Allen Brain Atlas, Brain Explorer 2). The overlaid circle corresponds to the specified FOV (∅ 5 mm). MOp: primary somatomotor, MOs: secondary somatomotor, SSp-ul: primary somatosensory upper limb, SSp-ll: primary somatosensory lower limb, SSp-un: primary somatosensory unassigned, SSp-tr: primary somatosensory trunk, SSp-bfd: primary somatosensory barrel field, PTLp: posterior parietal association, RSP: retrosplenial, VISam: anteromedial visual, VISpm: posteromedial visual, VISp: primary visual. (B) Schematic of a neuron (approximate soma diameter, 10 μm), and a diffraction limited point spread function (PSF) corresponding to NA 0.6 and 970 nm. Subcellular resolution implies that the PSF must be contained within a single neuron. (C) Key specifications for the mesoscope. (D) Imaging parameters used in the experiments. In all cases a fast resonant scan (time per scan line, 42 µs; 24 kHz line rate) is made over patches 608 µm wide. The time per patch is 42 µs x number of lines. The entire FOV can be covered in nine stripes. A slow (0.7 Hz), high-resolution scan with 3500 lines per stripe was used to cover a large portion the FOV (i). A fast (4.3 Hz), low-resolution scan across the entire scan range has up to 500 lines per stripe (iv). More typical scans will involve sampling multiple smaller patches at high resolution and frame rates (v). See implementation for discussion of the scanning patterns. (E) Schematic showing a typical use-case. The entire imaging volume is a cylinder with a 5 mm diameter and 1 mm depth. The fast scan is made over patches in different parts of the volume.