Journal: STAR Protocols

Article Title: Construction of a high-NFOM multiphoton microscope with large-angle resonant raster scanning

doi: 10.1016/j.xpro.2022.101330

Figure Lengend Snippet: Schematics of LAORS system with Zemax ray tracing Input beam: optical pulses from a pulsed laser source; resonant mirror: CRS 4 kHz (Cambridge Technology, USA); galvanometer mirror: 8320K (Cambridge Technology, USA); scan lens: LSM05-BB, effective focal length (EFL) of 110 mm (Thorlabs, USA); custom tube lens: a combination of 3 plano-convex lenses (Edmund Optics: 86-925, EFL of 500 mm), combined EFL of 166.7 mm; objective lens: Olympus XLUMPlanFl, 20×/0.95W. Note that multiple colors are utilized to better distinguish the rays under different scanning angles by the resonant and galvanometer scanning mirrors (see Figure 2 for more information on optical performance at different scanning angles).

Article Snippet: Refer to depicting the essential elements of a large-angle optical raster scanning (LAORS) system, primarily comprising: a. a typical pulsed laser source with an adequate repetition rate (Fidelity 2, Coherent, USA), b. a raster scanning unit with a resonant scanner (CRS 4 kHz, driver: 311-149887) for fast X-axis and a galvanometer scanner (8320K, driver: MicroMax 671) for slow Y-axis (both from Cambridge Technology, USA), c. a general scan lens (LSM05-BB, Thorlabs, USA) with an effective focal length (EFL) of 110 mm, d. a dedicated tube lens was custom designed combining three plano-convex lenses (Edmund Optics: 86-925), each with clear aperture and EFL of 73.5 mm and 500 mm, respectively, resulting in a combined EFL of 166.7 mm, e. a high-NA and low magnification objective lens (Olympus XLUMPlanFl, 20×/0.95W) with an EFL of 9 mm.

Techniques:

Journal: STAR Protocols

Article Title: Construction of a high-NFOM multiphoton microscope with large-angle resonant raster scanning

doi: 10.1016/j.xpro.2022.101330

Figure Lengend Snippet: Block diagram of the proposed control and data acquisition system The system comprises of- a computer: with i7-9800X 8-core processor and Nvidia Quadro RTX 8000 for CUDA-acceleration; a multifunction I/O card: PCIe-6341 (National Instruments Corporation, USA); a digitizer: ATS9440 (Alazar Technologies Inc., Canada); a 70 MHz pulsed laser source: Fidelity 2 Yb-Fiber Laser (Coherent Inc., USA); a resonant scanning system: CRS 4 kHz (driver: 311-149887), a galvanometer scanning system: 8320K (driver: MicroMax 671), Cambridge Technology, USA; a 16-bit digital to analog converter (DAC): 6757 (Cambridge Technology, USA); a voltage buffer: 50LD (Thorlabs, USA); a power splitter: ZFRSC-42-S+ (Mini-Circuits, USA); a high voltage power supply: C9525 (Hamamatsu Photonics, Japan); a transimpedance amplifier: C6438-01 (Hamamatsu Photonics, Japan); a multi-axis stage controller: SHOT-304GS, and three linear stages: 2 × TSDM40-15X and 1 × SGSP80-20ZF (Sigma Koki, Japan); a software: C++ written custom application.

Article Snippet: Refer to depicting the essential elements of a large-angle optical raster scanning (LAORS) system, primarily comprising: a. a typical pulsed laser source with an adequate repetition rate (Fidelity 2, Coherent, USA), b. a raster scanning unit with a resonant scanner (CRS 4 kHz, driver: 311-149887) for fast X-axis and a galvanometer scanner (8320K, driver: MicroMax 671) for slow Y-axis (both from Cambridge Technology, USA), c. a general scan lens (LSM05-BB, Thorlabs, USA) with an effective focal length (EFL) of 110 mm, d. a dedicated tube lens was custom designed combining three plano-convex lenses (Edmund Optics: 86-925), each with clear aperture and EFL of 73.5 mm and 500 mm, respectively, resulting in a combined EFL of 166.7 mm, e. a high-NA and low magnification objective lens (Olympus XLUMPlanFl, 20×/0.95W) with an EFL of 9 mm.

Techniques: Blocking Assay, Control, Software

Journal: STAR Protocols

Article Title: Construction of a high-NFOM multiphoton microscope with large-angle resonant raster scanning

doi: 10.1016/j.xpro.2022.101330

Figure Lengend Snippet: Entering a few important settings prior to imaging (A) The Settings GUI with multiple panels for resonant & galvanometer scanners, 3D stage, PMT high voltage (HV) supply, and additional parameters. (B and C) Calibration of resonant scanning angle, where B shows a simple method to estimate a scanning angle (optical) and C depicts a linear fitted plot for voltage vs optical scanning angle.

Article Snippet: Refer to depicting the essential elements of a large-angle optical raster scanning (LAORS) system, primarily comprising: a. a typical pulsed laser source with an adequate repetition rate (Fidelity 2, Coherent, USA), b. a raster scanning unit with a resonant scanner (CRS 4 kHz, driver: 311-149887) for fast X-axis and a galvanometer scanner (8320K, driver: MicroMax 671) for slow Y-axis (both from Cambridge Technology, USA), c. a general scan lens (LSM05-BB, Thorlabs, USA) with an effective focal length (EFL) of 110 mm, d. a dedicated tube lens was custom designed combining three plano-convex lenses (Edmund Optics: 86-925), each with clear aperture and EFL of 73.5 mm and 500 mm, respectively, resulting in a combined EFL of 166.7 mm, e. a high-NA and low magnification objective lens (Olympus XLUMPlanFl, 20×/0.95W) with an EFL of 9 mm.

Techniques: Imaging

Journal: STAR Protocols

Article Title: Construction of a high-NFOM multiphoton microscope with large-angle resonant raster scanning

doi: 10.1016/j.xpro.2022.101330

Figure Lengend Snippet:

Article Snippet: Refer to depicting the essential elements of a large-angle optical raster scanning (LAORS) system, primarily comprising: a. a typical pulsed laser source with an adequate repetition rate (Fidelity 2, Coherent, USA), b. a raster scanning unit with a resonant scanner (CRS 4 kHz, driver: 311-149887) for fast X-axis and a galvanometer scanner (8320K, driver: MicroMax 671) for slow Y-axis (both from Cambridge Technology, USA), c. a general scan lens (LSM05-BB, Thorlabs, USA) with an effective focal length (EFL) of 110 mm, d. a dedicated tube lens was custom designed combining three plano-convex lenses (Edmund Optics: 86-925), each with clear aperture and EFL of 73.5 mm and 500 mm, respectively, resulting in a combined EFL of 166.7 mm, e. a high-NA and low magnification objective lens (Olympus XLUMPlanFl, 20×/0.95W) with an EFL of 9 mm.

Techniques: Software