Hi everyone,
Here I am working on neurodegeneration model and I am in a need to visualize apoptosis in early as well as late phase. I am aware of TUNEL assay, but I am more interested in experimenting acridine orange/ethidium bromide staining to see apoptosis. My experimental model is "in vivo", how I am supposed to perform this staining experiment on brain tissue?
As I am naive for this experiment I would love to get elaborated explanation.
Thanks in advance!!!
Hi I am sending EtBr/AO staining protocol used in my lab
First make cell suspension in PBS.
Now mixed the cell suspension with EtBr/AO staining solution (1 mg/ml of AO in PBS + 1 mg/ml of EtBr in PBS) and incubate for 2 minute on ice.
Now mount the mixture on slide and visualize the cells under fluorescence microscope.
Capture the images using TRITC,FITC and & DIC. After merging the image using any software u can differentiate the apoptotic cells.
Hi I am sending EtBr/AO staining protocol used in my lab
First make cell suspension in PBS.
Now mixed the cell suspension with EtBr/AO staining solution (1 mg/ml of AO in PBS + 1 mg/ml of EtBr in PBS) and incubate for 2 minute on ice.
Now mount the mixture on slide and visualize the cells under fluorescence microscope.
Capture the images using TRITC,FITC and & DIC. After merging the image using any software u can differentiate the apoptotic cells.
Hello, in my experiment I use ao to count live cell. And you can use FACS to see apoptosis, by using the same protocol mentioned above or just follow the data sheet provided with reagent.
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