Method: Live Cell Imaging
About: Simply saying Live Cell Imaging is a technique which utilize advanced microscopy methods to analyse, observe and record behaviour of the living organisms, cells, organelles and even single proteins. Whole cell organisms (and organelles) are usually visualized with phase contrast and differential interference contrast (DIC) techniques which allow to investigate theri optical properties, dynamics, and morphology. When single proteins are investigated, a fluorescent fusion of the protein of interest has to be expressed in the cell to visualize it (fluorescent and confocal microscopy).
What: U2OS cells (osteosarcoma) expressing RFP-H2B (Red Fluorescent Protein - Histone 2B fusion, DNA visualization) and GFP-centrin (Green Fluorescent Protein - Centrin fusion, Centrosome visuzalization).
How: Live cell imaging using a Delta Vision Microscope. Cells were followed for 24h.
What you can see on the picture below is a few U2OS cells expressing histone H2B fused to RFP (red coulour) and centrin fused to GFP (green colour, see picture below).
Histone H2B and centrin are DNA binding/packing and centrosome proteins, respectively (for more information click here Histones, DNA packing proteins and here Fluorescent Micriscopy and Centrosome Staining). DNA will appears as a round shape like structure where centrosome is just a distinct spot (you may see it only in some of the cells, because not all of them express GFP-centrin or centrosome not always stay in focus of the camera). You will see few cells dividing (see below, an example of cell that will divide during the film is indicated with blue frame).
And now the movie:
Movie taken by Helen Dodson, PhD
I hope you enjoy it:)
Have a nice day,
Maciek GGSTEAM
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