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Table 1. Laser light sources, dichroic mirrors and filters used to visualize photosynthetic pigments/complexes or dyes in the Nikon PCM2000 confocal microscope The dichroic mirror is used to separate the laser excitation light from the fluorescence. Its cut-off wavelength should be longer than the laser excitation laser wavelength, but shorter than the emission maximum of the fluorophore, if possible. An emission filter is used to select the fluorescence wavelengths to be observed.
| Pigment/complex |
Organism |
Laser |
Wavelength |
Dichroic |
Emission filter |
Transmission |
|
| Intact phycobilisomes |
Cyanobacteria |
Red HeNe |
633 nm |
650 nm |
Schott RG665 |
>665 nm |
| Phycocyanin |
Cyanobacteria |
Red HeNe |
633 nm |
650 nm |
Interference edge filter |
>650 nm |
| Phycoerythrin |
Cyanobacteria |
Green HeNe |
543 nm |
565 nm |
Interference band-pass |
(605±15) nm |
| Photosystem II/IsiA |
Cyanobacteria |
Argon |
457 nm |
475 nm |
Schott RG665 |
>665 nm |
| Photosystem II/LHCII |
Plants |
Argon |
488 nm |
505 nm |
Schott RG665 |
>665 nm |
| GFP |
Any |
Argon |
488 nm |
505 nm |
Interference band- pass |
(515±15) nm |
| BODIPY FL C12 |
Any |
Argon |
488 nm |
505 nm |
Interference band-pass |
(515±15) nm |
|
|
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