Measurement of green fluorescent protein concentration in single cells by image analysis |
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| Authors: | Furtado Agnelo Henry Robert |
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| Institution: | Cooperative Research Centre for Molecular Plant Breeding, Centre for Plant Conservation, Genetics, Southern Cross University, Military Road, Lismore, NSW 2480, Australia. |
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| Abstract: | The gene encoding the green fluorescent protein (GFP) has been widely used in studies of gene expression. The GFP can be detected nondestructively in living cells or tissues by the green fluorescence of the protein under blue light. Solutions of enhanced GFP (EGFP) of known concentration were filled in glass capillaries and used to calibrate a method for quantitative determination of EGFP or GFP-S65T in plant cells. Images captured by a digital camera were analyzed to determine the linear range for measurement of EGFP expression. The value of the method was illustrated by analysis of the relative levels of GFP expression under control of different promoters in aleurone cells of barley. |
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| Keywords: | Image analysis GFP Fluorescence Promoter analysis Aleurone |
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