The aforementioned transgenic larvae from the Cu2+-stressed group and the control group (100 larvae/sample) at 68 hpf were dissociated separately into single cells as mentioned above, followed by incubation with the Fluorometric Intracellular ROS kit (Deep Red Fluorescence, Sigma-Aldrich, Cat# MAK142) according to the manual instructions. Finally, the isolated and stained cells were analyzed by FACS (CytoFLEX S, Beckman Coulter, USA). In this study, the FACS gate was determined based on the background fluorescence using a sample that contains the embryonic cells without the ROS Deep Red Fluorescence reagent staining.
Fluorometric intracellular ros kit deep red fluorescence
Manufactured by Merck Group
Sourced in Germany
The Fluorometric Intracellular ROS kit (Deep Red Fluorescence) is a lab equipment product designed to detect and measure reactive oxygen species (ROS) within cells. This kit utilizes a deep red fluorescent probe that specifically binds to ROS, allowing for quantification of intracellular ROS levels.
Automatically generated - may contain errors
2 protocols using fluorometric intracellular ros kit deep red fluorescence
1
Quantifying Cellular ROS Levels in Transgenic Larvae
2
Quantifying Intracellular ROS Levels
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Intracellular ROS was detected using Fluorometric intracellular ROS kit (Deep red fluorescence, Sigma, Taufkirchen, Germany). The unbiased tracking software of ZEN 3.0 automatically detected and counted fluorescent cells. Microscopic settings were identical throughout the entire analysis of all images. Data are shown as number of cells divided by total number of cells counted. Cell death was detected by propidium iodide permeabilization. Fluorescence was detected using Axio Observer microscope and ZEN 3.0 (blue edition) software (Zeiss, Oberkochen, Germany) or using microplate reader NOVOstar (BMG Labtech, Offenburg, Germany).
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