Journal: Nature Protocols

Article Title: Design and construction of a multiwavelength, micromirror total internal reflectance fluorescence microscope

doi: 10.1038/nprot.2014.155

Figure Lengend Snippet: Figure 3 | The components of the mmTIRF system and their assembly. (a) A diagram of the mmTIRF system (Mad City Labs) showing how each of the components interface. The system is designed around an objective holder fixed to a metal plate. A Nano-View/M 200-3 embedded-style xy-micrometer stage is then attached to this same plate. The nanopositioner itself is recessed into the micropositioner. A z-positioner with a dampening mass and a clamp to hold the U-shaped microscope slide holder is secured to the top plate of the nanopositioner. (b) The assembled micromirror TIRF system showing the objective platform, stages, legs and micromirror platform assemblies.

Article Snippet: PAC061AR.14, PAC055AR.14 (2×) and PAC074AR.14) Long-pass dichroic mirrors for dual-view (DM1 and DM2; Chroma Technology, cat. no. DCXR (2×)) Broadband dielectric mirrors for dual-view (M4 and M5; Newport Corporation, cat. no. 20Z20BD.1 (2×)) Band-pass, long-pass and notch emission filters (EFs; e.g., Semrock cat. no. FF03-525/50-25 488-nm band-pass filter) High-speed filter wheel (FW; Sutter Instrument, Lambda 10-3) Single photon detection EM-CCD camera (Andor, iXon3 897) Alignment laser (JDS, cat. no. 1508-2) Precision optical rail for mounting lenses, dual-view, filter wheel and EM-CCD camera (Newport Corporation, cat. no. PRL3 (3×)) Rail carrier (Newport Corporation, PRC series) Stainless steel optical posts for mounting optical components, 1/2-inch diameter (Thorlabs, TR series posts) Post holders for mounting optical components, 1/2-inch diameter (Thorlabs, PH series post holders) Kinematic cage mounts for mounting xy-translation mounts containing doublet lenses (Thorlabs, cat. no. KC1-T (3×)) xy-translation mounts for housing doublet lenses (Thorlabs, cat. no. LM1XY (3×)) Solid aluminum bread board, 8 inch × 10 inch, with 1/4-inch-20 tapped holes on 1-inch centers for dual-view platform (Thorlabs, MB series) Lens tubes (Thorlabs, SM30 series) Post-mounted iris diaphragms (Thorlabs, cat. no. ID25 (4×)) Threaded iris diaphragms for mounting in lens tubes (Thorlabs, cat. no. SM1D25) Three axes micropositioner (Newport Corporation, cat. no. MT-XYZ) Adjustable height V-mount for mounting final focusing lens (Thorlabs, cat. no. VG100) Optical support rod for mounting EM-CCD camera (Newport Corporation, cat. no. 70) Rack-and-pinion rod clamp part of EM-CCD camera mount (Newport Corporation, cat. no. 370-RC) Large-angle bracket for mounting camera (Thorlabs, cat. no. AP90RT) Linear translation stages for fine translation of mounted camera (Newport Corporation, cat. no. 423) • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • © 20 14 N at u re A m er ic a, In c. A ll ri g h ts r es er ve d . 2322 | VOL.9 NO.10 | 2014 | nature protocols proceDure arrangement and alignment of excitation path and installation of filters, continuous attenuators and beam expanders ● tIMInG 2 weeks 1| Draw the layout of the components of the microscope on the optical table (Fig. 5).

Techniques: Microscopy

Journal: Nature Protocols

Article Title: Design and construction of a multiwavelength, micromirror total internal reflectance fluorescence microscope

doi: 10.1038/nprot.2014.155

Figure Lengend Snippet: Figure 5 | A diagram of the position of the components of the microscope on the optical table.

Article Snippet: PAC061AR.14, PAC055AR.14 (2×) and PAC074AR.14) Long-pass dichroic mirrors for dual-view (DM1 and DM2; Chroma Technology, cat. no. DCXR (2×)) Broadband dielectric mirrors for dual-view (M4 and M5; Newport Corporation, cat. no. 20Z20BD.1 (2×)) Band-pass, long-pass and notch emission filters (EFs; e.g., Semrock cat. no. FF03-525/50-25 488-nm band-pass filter) High-speed filter wheel (FW; Sutter Instrument, Lambda 10-3) Single photon detection EM-CCD camera (Andor, iXon3 897) Alignment laser (JDS, cat. no. 1508-2) Precision optical rail for mounting lenses, dual-view, filter wheel and EM-CCD camera (Newport Corporation, cat. no. PRL3 (3×)) Rail carrier (Newport Corporation, PRC series) Stainless steel optical posts for mounting optical components, 1/2-inch diameter (Thorlabs, TR series posts) Post holders for mounting optical components, 1/2-inch diameter (Thorlabs, PH series post holders) Kinematic cage mounts for mounting xy-translation mounts containing doublet lenses (Thorlabs, cat. no. KC1-T (3×)) xy-translation mounts for housing doublet lenses (Thorlabs, cat. no. LM1XY (3×)) Solid aluminum bread board, 8 inch × 10 inch, with 1/4-inch-20 tapped holes on 1-inch centers for dual-view platform (Thorlabs, MB series) Lens tubes (Thorlabs, SM30 series) Post-mounted iris diaphragms (Thorlabs, cat. no. ID25 (4×)) Threaded iris diaphragms for mounting in lens tubes (Thorlabs, cat. no. SM1D25) Three axes micropositioner (Newport Corporation, cat. no. MT-XYZ) Adjustable height V-mount for mounting final focusing lens (Thorlabs, cat. no. VG100) Optical support rod for mounting EM-CCD camera (Newport Corporation, cat. no. 70) Rack-and-pinion rod clamp part of EM-CCD camera mount (Newport Corporation, cat. no. 370-RC) Large-angle bracket for mounting camera (Thorlabs, cat. no. AP90RT) Linear translation stages for fine translation of mounted camera (Newport Corporation, cat. no. 423) • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • © 20 14 N at u re A m er ic a, In c. A ll ri g h ts r es er ve d . 2322 | VOL.9 NO.10 | 2014 | nature protocols proceDure arrangement and alignment of excitation path and installation of filters, continuous attenuators and beam expanders ● tIMInG 2 weeks 1| Draw the layout of the components of the microscope on the optical table (Fig. 5).

Techniques: Microscopy

Journal: Nature Protocols

Article Title: Design and construction of a multiwavelength, micromirror total internal reflectance fluorescence microscope

doi: 10.1038/nprot.2014.155

Figure Lengend Snippet: Figure 7 | The components and layout of the emission path optics. (a) Location of the micromirrors, iris and the 45° mirror under the microscope objective. (b) Mounting of the EM-CCD camera. (c) The dual-view optics as seen looking toward the EM-CCD camera. (d) The dual-view optics as viewed looking toward the microscope objective.

Article Snippet: PAC061AR.14, PAC055AR.14 (2×) and PAC074AR.14) Long-pass dichroic mirrors for dual-view (DM1 and DM2; Chroma Technology, cat. no. DCXR (2×)) Broadband dielectric mirrors for dual-view (M4 and M5; Newport Corporation, cat. no. 20Z20BD.1 (2×)) Band-pass, long-pass and notch emission filters (EFs; e.g., Semrock cat. no. FF03-525/50-25 488-nm band-pass filter) High-speed filter wheel (FW; Sutter Instrument, Lambda 10-3) Single photon detection EM-CCD camera (Andor, iXon3 897) Alignment laser (JDS, cat. no. 1508-2) Precision optical rail for mounting lenses, dual-view, filter wheel and EM-CCD camera (Newport Corporation, cat. no. PRL3 (3×)) Rail carrier (Newport Corporation, PRC series) Stainless steel optical posts for mounting optical components, 1/2-inch diameter (Thorlabs, TR series posts) Post holders for mounting optical components, 1/2-inch diameter (Thorlabs, PH series post holders) Kinematic cage mounts for mounting xy-translation mounts containing doublet lenses (Thorlabs, cat. no. KC1-T (3×)) xy-translation mounts for housing doublet lenses (Thorlabs, cat. no. LM1XY (3×)) Solid aluminum bread board, 8 inch × 10 inch, with 1/4-inch-20 tapped holes on 1-inch centers for dual-view platform (Thorlabs, MB series) Lens tubes (Thorlabs, SM30 series) Post-mounted iris diaphragms (Thorlabs, cat. no. ID25 (4×)) Threaded iris diaphragms for mounting in lens tubes (Thorlabs, cat. no. SM1D25) Three axes micropositioner (Newport Corporation, cat. no. MT-XYZ) Adjustable height V-mount for mounting final focusing lens (Thorlabs, cat. no. VG100) Optical support rod for mounting EM-CCD camera (Newport Corporation, cat. no. 70) Rack-and-pinion rod clamp part of EM-CCD camera mount (Newport Corporation, cat. no. 370-RC) Large-angle bracket for mounting camera (Thorlabs, cat. no. AP90RT) Linear translation stages for fine translation of mounted camera (Newport Corporation, cat. no. 423) • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • © 20 14 N at u re A m er ic a, In c. A ll ri g h ts r es er ve d . 2322 | VOL.9 NO.10 | 2014 | nature protocols proceDure arrangement and alignment of excitation path and installation of filters, continuous attenuators and beam expanders ● tIMInG 2 weeks 1| Draw the layout of the components of the microscope on the optical table (Fig. 5).

Techniques: Microscopy