welcome again in Meet the chickens section. Today a specific labelling of chromosomes (actually sister chromatids) will be presented.
As you probably remember from previous post (Meet the chickens – chook chromosomes at hand), there is a way to examine karyotype (set of chromosomes) of chicken DT40 cell line (or any other cell line:). In this post we will combine this technique with a labelling procedure. This protocol will give us oportunity to see differential staining of sister chromatids.
Ok lets go.
In this protocol a nucleotide derivative, called BrdU (5-bromo deoxyuridine) is used to labell DNA for two cell cycles (see the picture below).
About: simply saying flow cytometry is a technique where properties of microscopic particles (such as cells or chromosomes) is examined. Flow cytometry is a very powerful method because it can measure many particles in the same time (up to thousands). Additionally, flow cytometry analysis is multiparametric, so different properties of particles can be measured at once.
What: Analysis of cell cycles distribution in different populations of cells.
How: By flow cytometry.
Ok, so let’s start with some short background on how the flow cytometer looks like (see picture below)
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