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As for the apoptosis experiment, first of all, we are going to discuss the advantages of this kind of experiment. Firstly, it can be used to study on the different types of pathological specimens, such as, tumor cells and tissues. Secondly, it also can be used in clinical diagnosis, drug development, bio-products development and cancer chemotherapy, as well as the gene treatment of tumor. And the last one is to diagnosis disease at the earlier stage.
Double labeling fluorescent probe
In this experiment, we choose 1μg / ml three harringtonine HT make HL-60 cells apoptosis in vitro. And then some cells are apoptosised. And then we use the Hoechst33342 and propidium iodide,PI to stain cells, which can separate the dead cells and live cells. HT works well in acute monocytic leukemia and according to the research, HT can lead HL-60 cells to apoptosis and show the features of apoptosis. What’s more, membrane is a kind of selective biofilm. General biofuels, such as, PI, can not go through it. However, when the cell died, the membrane is not complete, so PI can get through it to stain it.
Annexin V
Phosphatidylserine, PS is located inside of the membrane. At the early stage of apoptosis, PS can get out of the membrane and exist on the surface. Annexin-V is a pro-phospholipid protein, which molecular weight is 35~36 KD. It can combine with PS. Using the Annexin-V, which has been marked by FITC, PE or biotin as a fluorescent probe to detect the apoptosis with the help of flow cytometry or fluorescence microscopy. Propidine iodide---PI is a nucleic acid dye, which cannot pass through the membrane, however, the apoptosis cells will let the PI in. So the Annexin-V with the help of PI can be used to detect the live cells and dead cells.
These arE theories about different methods to analyze apoptosis.
Double labeling fluorescent probe
In this experiment, we choose 1μg / ml three harringtonine HT make HL-60 cells apoptosis in vitro. And then some cells are apoptosised. And then we use the Hoechst33342 and propidium iodide,PI to stain cells, which can separate the dead cells and live cells. HT works well in acute monocytic leukemia and according to the research, HT can lead HL-60 cells to apoptosis and show the features of apoptosis. What’s more, membrane is a kind of selective biofilm. General biofuels, such as, PI, can not go through it. However, when the cell died, the membrane is not complete, so PI can get through it to stain it.
Annexin V
Phosphatidylserine, PS is located inside of the membrane. At the early stage of apoptosis, PS can get out of the membrane and exist on the surface. Annexin-V is a pro-phospholipid protein, which molecular weight is 35~36 KD. It can combine with PS. Using the Annexin-V, which has been marked by FITC, PE or biotin as a fluorescent probe to detect the apoptosis with the help of flow cytometry or fluorescence microscopy. Propidine iodide---PI is a nucleic acid dye, which cannot pass through the membrane, however, the apoptosis cells will let the PI in. So the Annexin-V with the help of PI can be used to detect the live cells and dead cells.
These arE theories about different methods to analyze apoptosis.