Method to be used in fluorescence microscopy
View Patent ↗A method using fluorescence microscopy for image evaluation using a laser scanning microscope in which an at least partially spectrally resolved detection of the fluorescence spectrum occurs. Reference spectra are used for spectral demixing. Temporally and/or spectrally variable dyes and/or dye combinations are employed for recording of the reference spectra. Finally, the recorded reference spectra are inspected for image evaluation.
1. A method using fluorescence microscopy for image evaluation, the method comprising the steps of:
using a laser scanning microscope in which an at least partially spectrally resolved detection of the fluorescence spectrum occurs;
using reference spectra for spectral demixing;
employing temporally and/or spectrally variable dyes and/or dye combinations for recording of the reference spectra; and
demixing the recorded reference spectra for image evaluation.
2. The method according to claim 1 wherein the method records organic processes.
3. The method according to claim 1 wherein the method records intracellular processes.
4. The method according to claim 2 wherein the method records intercellular processes.
5. The method according to claim 1 wherein the method records cells and/or cell populations.
6. The method according to one claim 1 in which recording of reference spectra of photoconvertible dyes occurs.
7. The method according to one claim 1 in which recording of reference spectra of photoactivatable dyes occurs.
8. The method according to claim 1 in which recording of reference spectra of indicator dyes occurs.
9. The method according to claim 1 in which recording of reference spectra of dyes occurs that change their spectra dynamically based on intracellular processes.
10. The method according to one claim 1 in which recording of reference spectra of dyes occurs with a different rise in fluorescence intensity.
11. The method according to claim 1 in which recording of reference spectra of the fluorescing protein Kaede occurs.
12. The method according to one claim 1 in which recording of reference spectra of PA-GFP occurs.