I am reading a text about cell cycle and I have a doubt. I have read: "The fluorescence intensity of sub G0 cell fraction representes the apoptotic cell population".
In this text it is used iodure propidium to analyse the cell cycle. However, I am not completely sure but I think that only with iodure propidium the G0 phase is not visible (the visible phase is G0/G1).
Furthermore, in my mind G0 phase include other types of cells but not only apoptotic ones.
For this technique, cells are permeabilised and then propidium iodide is used to determine DNA content of the cells. Thus, cells in the G0/G1 phase will have equivalent amounts of DNA since their DNA is not being replicated. Cells in the S phase will have increasing amounts of DNA and those in the G2 phase will have duplicated the amount of DNA present.
Since apoptosis involves DNA fragmentation, these fragments are leaked out of the cell when permeabilised, so a fluorescent signal will be observed for lower DNA contents than in cells in the G0/G1 phase (what they call this sub G0/G1 fraction). Check figure of Sub G1 Analysis at: http://www.icms.qmul.ac.uk/flowcytometry/uses/apoptosis/dnafragmentation/
G0 refers to cells in a quiescent state. I'm not an expert on the subject, but I'm pretty sure they can also undergo apoptosis.
For this technique, cells are permeabilised and then propidium iodide is used to determine DNA content of the cells. Thus, cells in the G0/G1 phase will have equivalent amounts of DNA since their DNA is not being replicated. Cells in the S phase will have increasing amounts of DNA and those in the G2 phase will have duplicated the amount of DNA present.
Since apoptosis involves DNA fragmentation, these fragments are leaked out of the cell when permeabilised, so a fluorescent signal will be observed for lower DNA contents than in cells in the G0/G1 phase (what they call this sub G0/G1 fraction). Check figure of Sub G1 Analysis at: http://www.icms.qmul.ac.uk/flowcytometry/uses/apoptosis/dnafragmentation/
G0 refers to cells in a quiescent state. I'm not an expert on the subject, but I'm pretty sure they can also undergo apoptosis.
Hi, Guan, as G0 one can define non-cycling cells (cell which exit cell cycle) and undergo differentiation. This cells, usually, have similar DNA contents as G1 cells, but on the flow cytometry histogram peak may shift to "lower stain". This is not because of teh less DNA contents, but because of less DNA stain-ability. Ea in the case of differentiated cells chromatin was more condense to compare with G1 cells, so, it have lees ability to bind propidium iodine. In many case y differentiated cells can more easy undergo apoptosis, In that vase you can see on flow cytometry some peak corresponding to apoptotic cells, but this is not G0 cells. You can look for some of my paper about G0. Good luck!
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