A549 cells were collected from growth media after treating with 70 pM Au NRs for 4h and washed three times with 0.01 M PBS. The cells were incubated for 30 min under growth conditions with 500 μL of prewarmed medium containing 2 μmol/L LysoSensor Green DND-189 dye (Invitrogen). After washing, the cells were resuspended in 0.01 M PBS and immediately analyzed by an Attune® acoustic focusing cytometer (Applied Biosystems, Life Technologies, Carlsbad, CA). The cytometer was purchased jointly with the Nanotechnology lab for bioapplications, and was installed by Life Technologies Corp. in the National Center for Nanoscience and Technology, China. The green fluorescence was collected within 1 min from a population of 20 000 cells.
Lysosensor green dnd 189 dye
Manufactured by Thermo Fisher Scientific
Sourced in United States
LysoSensor Green DND-189 dye is a fluorescent probe designed for labeling acidic compartments within living cells. It exhibits a pH-dependent shift in fluorescence intensity, allowing its use as a tool for monitoring intracellular pH levels.
Automatically generated - may contain errors
2 protocols using lysosensor green dnd 189 dye
1
Lysosomal Labeling of Au NRs-Treated A549 Cells
2
Measuring Lysosomal pH and Acidic Particles
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Acidic particles in cells were measured using LysoTracker™ Deep Red dye (Invitrogen, USA) and Lysosomal pH was measured using LysoSensor™ Green DND-189 dye (Invitrogen, USA). Cells were cultured in ibidi glass slides and treated with respective compounds. After incubation, the media was replaced with media containing the respective dye, along with Hoesht 33342 dye (Invitrogen, USA) to stain the nucleus. Samples were imaged using Ziess LSM900 Confocal Microscope. Images were analyzed using Fiji. Signal was quantified per field of view and normalized to Hoesht.
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