Lifetime Flow Cytometry
SETA BioMedicals has developed some proprietary fluorescent dyes, tandems, and labels (fluorochromes) optimized to enable robust, fluorescence lifetime (FLT)-based, single- and multiple-cell analyses with Single Cell Fluorescence Lifetime Flow Cytometry (SC-FLFC) or High-Throughput Fluorescence Lifetime Imaging Flow Cytometry (HTFLIM-FC) [1]. These FLT-based reporters span the full spectral range from UV to near-IR, with luminescent lifetimes ranging from ps to tens of ns, and are compatible with the Danube Fluorescence Lifetime Flow Cytometer (Kinetic River Corp., USA) [2], Swabian Instrument FLFC [3], and other non-commercial FLT and FLIM FCs.
Excitation
laser line
[nm] |
Free dye / Ab conjugate‡
in phosphate buffer pH 7.4 |
Environment
sensitive
FLT |
λmax Ab
[nm] |
λmax Fl
[nm] |
Brightness at the excitation laser line
[M–1cm–1] |
FLT†
[ns] |
|
380
|
Seta-375 / IgG (D/P = 5)
|
No
|
372 / 372
|
476 / 480
|
20,500 / 50,000
|
3.2 / 3.5
|
|
SeTau-380 / BSA
|
No
|
381 / 383
|
480 / 468
|
900 / 3,000
|
32.5 / 30.6
|
|
405
|
SeTau-405 / IgG (D/P = 2)
|
No
|
405 / 405
|
518 / 518
|
11,000 / 6,900
|
9.3 / 8.9
|
|
436
|
SeTau-425 / BSA (D/P = 2)
|
n.d.*
|
425 / 427
|
545 /532
|
1,500 / 1,000
|
26.2 / n.d.*
|
|
Seta-470 / Glycerol:Water 10 cP, +65 °C / 700 cP, -6 °C
|
Yes, Viscosity
|
460 / 460 / 460
|
521 / 525 / 525
|
560 / n.d.*/ n.d.*
|
0.21 / 1.7 / 17.3
|
|
488
|
SeTau-488 / IgG (D/P = 2)
|
Yes
|
486 / 484
|
532 / 530
|
22,800 / 71,600
|
1.2 / 2.3
|
|
SeTau-500 / IgG (D/P = 2)
|
Yes
|
502 / 500
|
568 / 563
|
31,200 / 53,100
|
1.7 / 2.6
|
|
Seta-470 / Glycerol:Water 10 cP, +65 °C / 700 cP, -6 °C
|
Yes, Viscosity
|
460 / 460 / 460
|
521 / 525 / 525
|
560 / n.d.*/ n.d.*
|
0.21 / 1.7 / 17.3
|
|
532
|
Seta-555 / (D/P = 2)
|
No
|
555/ 556
|
570/ 571
|
3,400 / n.d.*
|
0.2 / n.d.*
|
|
594
|
Seta-633 / IgG (D/P = 2)
|
Yes
|
633 /637
|
644 /647
|
3,600 / 23,700
|
0.25 / 1.49
|
|
Seta-640 / IgG (D/P = 2)
|
Yes
|
636 / 648
|
646 / 656
|
4,600 / 19,900
|
0.7 / 2.7
|
|
635
|
Seta-650
|
No
|
647
|
665
|
50,300
|
1.0
|
|
Seta-660 / IgG (D/P = 2)
|
n.d.*
|
661 / 665
|
672 / 675
|
4,900 / 38,900
|
1.5 / n.d.*
|
|
SeTau-647 / IgG (D/P = 2)
|
No
|
649 / 648
|
695 / 694
|
74,000 / 128,600
|
3.2 / 3.1
|
|
650
|
Seta-650
|
No
|
651
|
671
|
65,400
|
1.0
|
|
Seta-660 / IgG (D/P = 2)
|
n.d.*
|
661 / 665
|
672 / 675
|
10,700 / 83,800
|
1.5 / n.d.*
|
|
SeTau-665 / IgG (D/P = 2)
|
No
|
664 / 662
|
712 / 716
|
47,900 / 54,200
|
3.1 / n.d.*
|
|
SeTau-670 / IgG (D/P = 4)
|
No
|
673 / 672
|
694 / 693
|
39,600 /61,600
|
1.62 / 1.75
|
|
Square 650-pH at pH 5.6 / pH 9.0
|
Yes, pH§
|
653 / 535
|
671 / 663
|
20,900 / 260 (3,600 at λ* = 532 nm)
|
1.2 / 0.5
|
|
Square-660/Oligonucleotides
|
Yes
|
657/676
|
676/695
|
n.d.* / n.d.*
|
0.29/2.73
|
|
680
|
Seta-680
|
No
|
676
|
694
|
50,400
|
1.0
|
|
SeTau-680
|
No
|
683
|
705
|
122,300
|
2.9
|
|
Seta-670 / IgG (D/P = 5) / BSA (D/P = 1.5)
|
Yes
|
667 / 670 / 681
|
688 / 693 / 696
|
8,200 / 11,700 / 47,300
|
0.42 / 0.85 / 2.43
|
|
700
|
Zy7
|
No
|
749
|
774
|
8,400
|
0.57
|
|
750
|
Seta-750 / IgG (D/P = 2)
|
No
|
753 / 756
|
780 / 783
|
31,600 / 68,700
|
0.6 / n.d.*
|
|
780
|
Seta-780
|
No
|
777
|
807
|
2,600
|
0.6
|
† Acquired with ISS Chronos FD at 25 °C
‡ Fluorochrome conjugated to antibody (protein). D/P is dye-to-protein ratio
* Not determined
§ pH-sensitive dye, excitable at 650 nm under acidic pH and at 532 nm at basic pH, emitting at 660–670 nm under all conditions [4].
References
[1] Nat Commun 15, 7376 (2024).
[2] https://www.kineticriver.com/wp-content/uploads/2022/05/KRC-Danube-Data-Sheet.pdf
[3] https://www.swabianinstruments.com/applications/fluorescence-lifetime-flow-cytometry/
[4] Analytical Biochemistry, 2009, V390, 136-140.
Please contact us if you are interested in a sample for testing.
