Cells were imaged on an inverted Nikon Ti-E (Nikon, Melville, NY) with a Yokogawa CSU-X confocal scanning head (Yokogawa Electric, Tokyo, Japan) and laser merge model with 491, 561, and 642nm laser lines (Spectral Applied Research, Ontario, Canada). Images were collected on a Zyla 4.2 sCMOS Camera (Andor, Belfast, UK). Optogenetic recruitment utilized a Mosaic digital micromirror device (Andor) coupled to a 405-nm laser. A 60× 1.49 NA ApoTIRF oil immersion objective (Nikon) or a 60× 1.2 Plan Apo water (Nikon) objective was used to collect images. MetaMorph Automation and Image Analysis Software (Molecular Devices, Sunnvyale, CA) controlled all hardware.
1.49 na apotirf oil immersion objective
Manufactured by Nikon
The 60× 1.49 NA ApoTIRF oil immersion objective is a high-performance microscope objective designed for Total Internal Reflection Fluorescence (TIRF) microscopy. It has a numerical aperture (NA) of 1.49, which provides a high-resolution and high-contrast imaging capability. The objective is designed for use with oil immersion, which helps to maximize the light collection efficiency and optimize the optical performance.
Automatically generated - may contain errors
Lab products found in correlation
Zyla 4.2 scmos camera, by Oxford Instruments (1 mentions)
Mosaic digital micromirror device, by Oxford Instruments (1 mentions)
Metamorph automation and image analysis software, by Molecular Devices (1 mentions)
Csu x confocal scanning head, by Yokogawa (1 mentions)
A1r hd25 confocal microscope, by Nikon (1 mentions)
2 protocols using 1.49 na apotirf oil immersion objective
1
Confocal Microscopy and Optogenetic Recruitment
2
Confocal Imaging and Analysis of GUVs
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Imaging was conducted using a Nikon A1R-HD25 Confocal Microscope using a 60 × 1.49 NA APO TIRF oil immersion objective (Nikon Instruments, Melville, NY). Secondary analysis of GUVs was conducted with NIH ImageJ. After background subtraction, the total fluorescence intensity of individual GUVs was measured by analyzing the integrated density of each GUV membrane. A circle surrounding the perimeter of each GUV membrane was first measured and then subtracted from measurement of a circle drawn around the interior of each GUV membrane. The total fluorescence intensity of each GUV membrane was normalized to its diameter to account for any disparities in GUV size.
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