GoKawiilMicroscope hardware
A complete MOSAIC CAD model, bill of materials, and nearly 1,000 pages of installation, maintenance and operation instructions are available through a no-cost research license agreement with Howard Hughes Medical Institute (HHMI).
Excitation sources
Visible lasers 405 nm, 100 mW (TOPTICA, IBEAM-SMART-405-S-BZ), 445 nm, 100 mW (IBEAM-SMART-445-S-BZ), 488 nm, 500 mW (MPB, 2RU-VFL-P-500-488-B1R), 514 nm, 1,000 mW (MPB, 2RU-VFL-P-1000-514-B1R), 560 nm, 1,000 mW (MPB, 2RU-VFL-P-1000-560-B1R), 607 nm, 1,000 mW (MPB, 2RU-VFL-P-1000-607-B1R) and 642 nm, 2,000 mW (MPB, 2RU-VFL-P-2000-6224-B1R) were coaligned in a custom laser combiner and passed through an Acousto-Optic Tunable Filter (AOTF; AA Optoelectronic, AOTFnC-400.650-CPCh-TN) driven via a four-channel Multi-Purpose Digital Synthesizer (AA, MPDS4C-B66-22-52.111) which allowed rapid, independent control of their laser amplitudes. The output beam from the AOTF was then sent free-space to the downstream optics.
The femtosecond laser used for TPM (Coherent Chameleon LS II, tunable from 680 nm to 1,080 nm, peak power >3.5 W) was first corrected for group velocity dispersion (GVD) via a prism compressor (Thorlabs) before passing through an infrared Acousto-Optic Modulator System (AOM; AA, MT110-B50A1.5-IR-Hk driven via MPDS1C-B6-34-85.135) to control the output power delivered to the downstream optics.
Spatial light modulator
A sample-conjugate SLM (Meadowlark P1920-0635-HDMI, 1,920 × 1,152 pixels) served multiple roles in MOSAIC: generation of LLS and 3D-SIM excitation patterns, five-axis alignment of the LLS to the detection focal plane, laser blanking, excitation AO correction, 3D phase modulation, light-sheet collimation and chromatic correction, and patterned photoactivation.
Mask
A motorized custom glass wheel with a circumferential series of different photolithographically produced patterned metallic masks (Thorlabs) specific to different light sheets or SIM patterns was used to pass only the desired first-order diffracted light while blocking the unwanted zeroth and higher orders.
Dichroic stack
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