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Quad-band filters forĀ fastĀ switching to the Zyla Camera. High speed but risk of cross-talk.
Filter ID | Excitation Wavelength | Dichroic Mirror | Emission Filter (in quad) | Example Fluorophores |
---|---|---|---|---|
DAPI | 385 nm | 86012v2 Dapi/FITC/TxRed/Cy5 | ~ 455nm - 485nm | DAPI, Hoechst |
FITC | 470 nm | 86012v2 Dapi/FITC/TxRed/Cy5 | ~ 510nm - 545nm | GFP, FITC |
TxRed | 550 nm | 86012v2 Dapi/FITC/TxRed/Cy5 | ~ 590nm - 630nm | RFP, mCherry |
Cy5 | 635 nm | 86012v2 Dapi/FITC/TxRed/Cy5 | ~ 670nm - 740nm | Cy5 |
Sedat quad-band dichroic and single-band emission filters for slower switching. Lower speed but reduced cross-talk between channels.
Filter ID | Excitation Wavelength | Dichroic Mirror | Emission Filter (in wheel) | Example Fluorophores |
---|---|---|---|---|
DAPI | 385 nm | 89000 Sedat Quad | 435 - 485 | DAPI, Hoechst |
GFP | 470 nm | 89000 Sedat Quad | 500 - 550 | GFP, FITC |
RFP | 550 nm | 89000 Sedat Quad | 590 - 650 | RFP, mCherry |
Cy5 | 660 nm | 89000 Sedat Quad | 662 - 737 | Cy5 |
CFP | 470 nm | 89002 CFP/YFP | 460 - 500 | CFP |
eYFP | 500 nm | 89002 CFP/YFP | 520 - 550 | YFP |
iSIM Filters
The iSIM has the following quad-band dichroic and emission filter in its scan head:. The emission filter is Chroma part number 88100bs from Pinkel set 88000v2.
Laser | Dichroic Mirror (reflection bands) | Emission Filter | Example Fluorophores |
---|---|---|---|
405 | 400 - 410 | 430 - 485 | DAPI, Hoechst |
488 | 483 - 493 | 520 - 550 | GFP, FITC |
561 | 556 - 566 | 590 - 630 | RFP, mCherry |
640 | 635 - 645 | 680 - 740 | Cy5 |
The iSim has the following emission filters in Optospin Wheel 0
Position | Laser | Emission band | Example Fluorophores |
---|---|---|---|
1 | Any | 88000v2 Quad | Most |
2 | 405 | 410 - 510 | DAPI, Hoechst |
3 | 488 | 475 - 575 | GFP, FITC |
4 | 561 | 550 - 650 | tdTomato, TRITC |
5 | 561 | 555 - 705 | mCherry, Texas Red |
6 | 642 | 625 - 775 | Cy5 |
Camera Specifications
- Zyla 4.2 Plus specifications (Andor website). Note that the Zyla on GFP4 is a Camera Link model and not USB 3.0.
- Hamamatsu Orca-Flash4.0 V3 specifications (Hamamatsu website)