Protocols / How To: Confocal Microscopy

Before samples are screened by powerful lasers, which could bleach the GFP signal, the mercury based transmission light might be used for browsing the sample. After the right sample layer was focused, the source of lightening was switched to the lasers and the pinhole value needed to be adjusted. Three pictures were taken: one in the green channel, one in the red channel and one of in pseudo-transmission light. GFP molecules gave a green signal, while the chlorophyll visualized the chloroplasts as red dots. In order to compare the pictures, a scale bar was added in the pictures.