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Fluorescence microscopy
From 2008.igem.org
(New page: Fluorescence microscopy is a common technique for examining fluorophores (fluorescent molecules). In this case, it was used to show the expression level of two reporter genes, ''gfp'' and ...)
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Revision as of 14:09, 19 September 2008
Fluorescence microscopy is a common technique for examining fluorophores (fluorescent molecules). In this case, it was used to show the expression level of two reporter genes, gfp and mcherry. A light beam from a mecury vapour lamp is focused onto a dichroic mirror, which reflects a adjustable wavelength range onto the sample. This should be the optimum excition wavelength for the fluorophore being observed. In the case of GFP, this is blue light (~395nm). For mCherry, this is yellow light (~590nm). This 'excitation' light causes the fluorophore to emit its own wavelength (~510nm for GFP and ~615nm for mCherry) which is focused onto the camera. In this way, cells (or specific parts of cells) can be visualised depending on where the reporter protein is localised.
