After a 2-hour coculture of WI-38 cells with PKH67-labeled plasma EVs (derived from 100 µl plasma) and the No EV control, the culture medium was discarded, and the cells on the plate were washed twice with 500 µl HBSS (Gibco). The cells were fixed with 100 µl 4% PFA for 15 minutes at 37°C, then washed three times with HBSS. WI-38 cell membranes were stained with Wheat Germ Agglutinin (WGA, ThermoFisher Scientific) and washed twice with HBSS. The cells were permeabilized with 0.2% Triton-X-100 (MilliporeSigma) for 5 minutes, followed by cell nuclei staining with DAPI (MilliporeSigma). The cells were washed twice with HBSS. Immunofluorescence images were captured on an Olympus IX70 Inverted Phase Contrast DIC Fluorescence Microscope accompanied with X-Cite 120LED Boost High-Power LED illumination System (EXCELITAS Technologies) and analyzed with cellSens Standard Software (Olympus).
X cite 120led boost high power led illumination system
Manufactured by Excelitas
The X-Cite 120LED Boost is a high-power LED illumination system designed for laboratory applications. It provides a stable and uniform illumination source with a wide range of wavelength options. The product's core function is to deliver controlled and consistent light output for various microscopy, fluorescence, and imaging techniques.
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Lab products found in correlation
Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (2 mentions)
Wheat germ agglutinin wga, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Cellsens standard software, by Olympus (1 mentions)
Zen acquisition software, by Zeiss (1 mentions)
Lambda 10 3 optical filter changer, by Sutter Instruments (1 mentions)
Orca flash4.0 v3 digital cmos camera, by Hamamatsu Photonics (1 mentions)
Ix81 motorized inverted microscope, by Olympus (1 mentions)
2 protocols using x cite 120led boost high power led illumination system
1
Imaging Plasma EV Uptake in WI-38 Cells
2
Multicolor Fluorescence Microscopy Imaging
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Epifluorescent images were acquired with an Olympus ix-81 motorized inverted microscope with CellSens acquisition software v1.0 (Olympus, Center Valley, PA), with a manual Olympus stage and software-based z-step, using an X-Cite 120LED Boost High-Power LED illumination System (Excelitas Technologies, Waltham, MA). Excitation/emission wavelengths were controlled using Lambda 10-3 Optical Filter Changer (Sutter Instrument, Novato, CA) equipped with the following filter excitation wheels located at the light source: AT350/50x, ET402/15x, ET490/20x, ET555/25x, ET645/30x (Chroma, Bellows Falls, VT). The emission wheel located on the camera port contained the following: ET455/50m, ET525/36m, ET605/52m, ET705/72m (Chroma). Between both wheels contained the 89100bs dichroic (Chroma). An Olympus UPlanFL N 60x/1.25 Oil Iris objective lens was used for acquisition. Detector used was an ORCA-Flash4.0 V3 Digital CMOS camera (Hamamatsu, Hamamatsu City, Japan); 2 × 2 binning, 6.5µmx6.5µm pixel size, 2048 × 2048 chip size, with rolling shutter. Confocal images were captured on an LSM 880 with Airyscan with ZEN acquisition software (Zeiss, Jena, Germany). z-stacks were acquired and processed using the Airyscan Processing algorithm included in ZEN.
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