An Eclipse Ti inverted microscope (Nikon) with a TIRF system and iXon electron-multiplying charged-coupled device camera (Andor Technology) was used for imaging. TIRF microscopy was performed with a Nikon 100 × 1.49 numerical aperture oil-immersion TIRF objective, a TIRF illuminator, a Perfect Focus system, a motorized stage, and a U-N4S four-laser unit as a laser source (Nikon). The laser unit has the solid-state lasers for the 488 nm, 561 nm, and 640 nm channels and was controlled using a built-in acousto-optic tunable filter. The laser powers of 488 nm laser, 561 nm laser, and 640 nm laser were set to 5.2, 6.9, and 7.8 mW respectively measured with the field aperture fully opened. The 405/488/561/638 nm Quad TIRF filter set (Chroma Technology Corp.), along with supplementary emission filters of 525/50 m, 600/50 m, 700/75 m for 488 nm, 561 nm, 640 nm channels, respectively, were used. Images are acquired using the Nikon NIS-Elements software with the exposure time of 50 ms.
100 1.49 numerical aperture oil immersion tirf objective
Manufactured by Nikon
The Nikon 100 × 1.49 numerical aperture oil-immersion TIRF objective is a high-performance microscope objective designed for Total Internal Reflection Fluorescence (TIRF) imaging. It has a magnification factor of 100× and a numerical aperture of 1.49, which enables it to capture high-resolution images with a shallow depth of field.
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Lab products found in correlation
Tirf system, by Oxford Instruments (2 mentions)
561 638 nm quad tirf filter set, by Chroma Technology (2 mentions)
Eclipse ti inverted microscope, by Nikon (2 mentions)
Nis elements software, by Nikon (2 mentions)
Perfect focus system, by Nikon (2 mentions)
Ixon electron multiplying charged coupled device camera, by Oxford Instruments (1 mentions)
Tirf illuminator, by Nikon (1 mentions)
2 protocols using 100 1.49 numerical aperture oil immersion tirf objective
1
Multicolor TIRF Microscopy Protocol
2
Multi-channel TIRF Microscopy Imaging
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An Eclipse Ti inverted microscope (Nikon) with a TIRF system and iXon EMCCD (Electron Multiplying Charged-Coupled Device) camera (Andor Technology) was used for imaging. TIRF microscopy was performed with a Nikon 100 × 1.49 numerical aperture oil-immersion TIRF objective, a TIRF illuminator, a Perfect Focus system, a motorized stage and a U-N4S 4-laser unit as a laser source (Nikon). The laser unit has the solid state lasers for the 488 nm, 561 nm and 640 nm channels, and was controlled using a built-in acousto-optic tunable filter (AOTF). The laser powers of 488 nm laser, 561 nm laser, and 640 nm laser were set to 5.2, 6.9, and 7.8mW measured with the field aperture fully opened. The 405/488/561/638 nm Quad TIRF filter set (Chroma Technology Corp., Rockingham, Vermont) along with supplementary emission filters of 525/50 m, 600/50 m, 700/75 m for 488 nm, 561 nm, 640 nm channel was used respectively. Images are acquired using the Nikon NIS-Elements software with the exposure time of 50 milliseconds.
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