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Turret Position | Filter Set | Excitation Filter | Dichroic Mirror | Emission Filter | Example Fluorophores |
|---|---|---|---|---|---|
1 | A | BP340-380 | RKP400 | LP425 | Hoechst/DAPI |
2 | I3 | BP450-490 | RKP510 | LP515 | FITC/GFP |
3 | N2.1 | BP515-560 | RKP580 | LP590 | Texas Red |
4 | BF | N/A | N/A | N/A | Transmitted Light |
5 | N/A | N/A | N/A | N/A | Empty |
6 | Pol | N/A | N/A | N/A | Polariser for DIC and polarised light microscopy |
Positions 4 to 6 are not used for fluorescence microscopyirrelevant for fluorescence.
Lasers and dichroic mirrors
The SPE3 has the following lasers. Note that it differs from the other SPEs in that it has a 561 nm laser instead of a 532 nm laser.
Laser Line | Example Fluorophores |
|---|---|
405 nm | DAPI, Hoechst |
488 nm | FITC, GFP |
561 nm | Texas Red, mCherry |
635 nm | Cy5, Alexa 647 |
Dichroic mirrors separate the excitation light from the emitted fluorescence. Without dichroic mirrors or other beam splitting technologies your image would be contaminated with laser light.
Dichroic Mirror | Description |
|---|---|
QD 405/488/561/635 | Quad band that reflects all four laser lines. This is the one you're most likely to use |
DD 488/561 | Double dichroic. Only reflects the 488 and 561 lines, which means the other two lines will not be blocked |
RT 30/70 | A beam splitter for back scatter or reflection imaging. Not useful for most fluorescence. |