Dissected larval pancreata were mounted in fluorescent mounting medium (Dako) and imaged with a Nikon A1-si Laser Scanning Confocal Head mounted on a TiE inverted microscope through a Plan Apo 20× VC (N.A. 0.75) (Nikon). Acquisition and analysis was performed using NIS-Elements software (Ver 3.22, build 710). Cell number was quantified by counterstaining nuclei with DAPI (0.2 μg/ml) and counted by using NIS-Elements software (Nikon).
Plan apo 20 vc
Manufactured by Nikon
Sourced in Japan
The Plan Apo 20× VC is a high-performance objective lens designed for use in various laboratory applications. It offers a magnification of 20X and features Vibration Compensation (VC) technology to provide stable and clear images. The lens is optimized for plan-apochromatic performance, ensuring accurate and uniform image quality across the entire field of view.
Automatically generated - may contain errors
4 protocols using plan apo 20 vc
1
Quantifying Pancreatic Cell Abundance
2
Quantitative imaging of larval pancreata
3
Fluorescence Imaging for Enzyme Kinetics
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Fluorescence images were obtained using an electron multiplying CCD camera (ImagEM Enhanced C9100-13, Hamamatsu Photonics K. K. (Japan)) and an inverted microscope (ECLIPSE Ti, Nikon Corp. (Japan)) equipped with 20× and 100× objective lenses (Plan Apo VC 20× (Nikon) and Plan Apo λ 100× oil (Nikon)). The 100× lens was used only for time-lapse imaging. Excitation and emission wavelengths were 470 nm and 535 ± 15 nm for fluorescein, 555 nm and 593 ± 20 nm for resorufin, and 395 nm and 480 ± 20 nm for 4-MU, respectively. For enzyme kinetic measurements and digital counting of enzyme, exposure time was 200 ms for fluorescein and 400 ms for resorufin. In dual-color enzyme assay, exposure time was 300 ms for fluorescein, 500 ms for resorufin and 100 ms for 4-MU. The fluorescence images were analyzed using ImageJ software (National Institutes of Health (USA)).
4
Multicolor Fluorescence Imaging Protocol
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Fluorescence images were acquired by confocal fluorescence microscope unit (AX, Nikon) equipped with a 20× dry objective lens (Plan Apo VC 20×), LASER unit (LUA‐S4), detector unit (DUX‐ST), and a motorized stage. The assay was performed using a solution containing sTM[6 (link)
] (10 µM) as the internal standard, and the focus was adjusted using its fluorescence. Images were acquired in tile scan mode with perfect focus. The excitation wavelength and emission filters used were DAPI (Ex. = 405 nm, Em. = 429–474 nm), FITC (Ex. = 488 nm, Em. = 500–550 nm) and Cy3 (Ex. = 561 nm, Em. = 570–616 nm), respectively. Images were taken as sequential acquisition mode.
] (10 µM) as the internal standard, and the focus was adjusted using its fluorescence. Images were acquired in tile scan mode with perfect focus. The excitation wavelength and emission filters used were DAPI (Ex. = 405 nm, Em. = 429–474 nm), FITC (Ex. = 488 nm, Em. = 500–550 nm) and Cy3 (Ex. = 561 nm, Em. = 570–616 nm), respectively. Images were taken as sequential acquisition mode.
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