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.

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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 565RedNone+++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* 505525Green
++
None
Use at 25-50 uM.
Annexin V-FITC494518Green++NoneUse according to PIS (test 10 ul Annexin V-FITC if signal is dim).
Acridine Orange500526Green+++NoneFor Jurkat cells suggest using 0.25 – 0.5 uM (test titration of the dye to ensure no overlap into the red channel)
Calcein AM494517Green++++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.