Tali® Dye Compatibility
The Tali® Image-Based Cytometer can be used to detect green and red fluorescence of a variety of dyes and stains. This table ranks the fluorescence intensity of various dyes when examined on the Tali® Image-Based Cytometer. Unless otherwise indicated, cells were labeled with the indicated fluorescent dye or stain at concentrations suggested in the Product Information Sheet and analyzed on the Tali® Image-Based Cytometer using the “Green+Red Setting” setting. In some cases, there was spectral overlap of the fluorescence emission (commonly known as bleedthrough) into the opposite channel. This information is also included in the table. |
Red Channel Dyes
| Dye, Product Tested | Excitation (nm) | Emission (nm) | Expected Emission Channel | Green Channel Signal | Red Channel Signal | Recommendations |
|---|---|---|---|---|---|---|
| 7-AAD (7-aminoactinomycin D) | 549 | 645 | Red | None | +++ | Use 1-2 ul per ml. |
| R-PE (R-phycoerythrin), CD3-RPE conjugate | 564 | 577 | Red | None | ++ | Test titration of antibody conjugate. |
| CellROX Orange Reagent | 545 | 565 | Red | None | +++ | Use according to PIS. |
| MitoTracker® Orange CM-H2TMRos | 551 | 575 | Red | None | ++ | Use according to PIS. |
| Ethidium homodimer-1 | 528 | 618 | Red | None | +++ | Use 1 uM. |
| Rhodamine Red-X anti-mouse secondary | 573 | 591 | Red | None | + | basically undetectable |
Green Channel Dyes
| Dye, Product Tested | Excitation (nm) | Emission (nm) | Expected Emission Channel | Green Channel Signal | Red Channel Signal | Recommendations |
|---|---|---|---|---|---|---|
| SYTOX® Green stain | 504 | 523 | Green | +++ | + | Use 12.5 nM. |
| LIVE/DEAD® Fixable Green Dead Cell Stain | 495 | 520 | Green | +++ | ++ | Spectral overlap into red channel at lowest concentration tested (0.25 µL/mL) |
| CellTrace™ Green | 492 | 517 | Green | +++ | + | Use at 50 nM–250 nM |
| CellEvent® Green reagent | 502 | 530 | Green | ++ | None | 10 µM final concentration |
| SYTO® 9 stain | 485 | 498/501 | Green | +++ | None | Use 100 nM |
| TO-PRO®-1 | 515 | 531 | Green | ++ | ++ | Due to spectral overlap can not be used for multiplexing experiments |
| FM® 1-43X | 510 | 626 | Green | +++ | +++ | Due to spectral overlap can not be used for multiplexing experiments |
| CellTracker™ Green CFDA-AM | 492 | 517 | Green | +++ | None | Use according to PIS. |
| 5-(and-6)-carboxy-2',7' -dichlorofluorescein diacetate (carboxy-DCFDA) *mixed isomers* | 505 | 525 | Green | ++ | None | Use at 25-50 uM. |
| Annexin V-FITC | 494 | 518 | Green | ++ | None | Use according to PIS (test 10 ul Annexin V-FITC if signal is dim). |
| Acridine Orange | 500 | 526 | Green | +++ | None | For Jurkat cells suggest using 0.25 – 0.5 uM (test titration of the dye to ensure no overlap into the red channel) |
| Calcein AM | 494 | 517 | Green | +++ | + | Use at 0.1 - 1 uM (test titration of dye to ensure no overlap into red channel). |
"+" undetectable to barely detectable on the histogram, but can see spots on the Tali images.
"++" can detect peak on histogram, but may be difficult to separate positive and negative peaks.
"+++" easily detectable peaks on histogram. Positive and negative populations can be easily resolved.
