Simultaneous imaging of confocal fluorescence and Raman spectrum

The advantage of confocal fluorescence microscopy is the ability to acquire high resolution images of fluorescent specimens non-invasively. On the other hand, the advantage of Raman microscopy is the ability to provide the chemical characteristics of specimens from spectroscopy. To obtain simultaneously high resolution images and chemical characteristics of specimens, fluorescence signals from stained cell and Raman spectrum from cell itself should be separated. By separating two kinds of signals, confocal fluorescence image and Raman spectrum are acquired simultaneously at the same position. In this paper, we demonstrate a confocal fluorescence microscopy combined with the Raman microscopy. And we propose a method that eliminates Raman spectrum of fluorophore itself from Raman spectrum of the stained cell and that obtains simultaneously confocal fluorescence image from stained cell and Raman spectrum of cell itself without replacing a cell.