415:, due to the progressive halving of CFSE fluorescence within daughter cells following each cell division. The only limitation is that CFSE at high concentrations can be toxic for cells. However, when CFSE labelling is performed optimally, approximately 7-8 cell divisions can be identified before the CFSE fluorescence is too low to be distinguished above the autofluorescence background. Thus CFSE represents an extremely valuable fluorescent dye for immunological studies, allowing lymphocyte proliferation, migration and positioning to be simultaneously monitored. By the use of fluorescent antibodies against different lymphocyte cell surface markers it is also possible to follow the proliferation behaviour of different lymphocyte subsets. In addition, unlike other methods, CFSE-labeled viable cells can be recovered for further analysis.
31:
198:
325:
435:, are division dependent. Sophisticated mathematical models have also been developed to analyse CFSE data and probe various aspects of immune responses. Furthermore, the use of CFSE has extended beyond the immune system, with the dye being used to monitor the proliferation of many other cell types such as
418:
Since the initial description of CFSE it has been used in thousands of immunological studies, an example of an early proliferation study in animals being described by Kurts et al. However, perhaps the most important CFSE investigations have been those demonstrating that many of the effector functions
462:
Detailed protocols are now available that can be used to label lymphocytes (and other cell types) with a high degree of reliability and precision. One of the most important parameters, however, is to ensure that the cell population being studied has not been too heavily labelled with CFSE, as such
380:
residues and other amine sources. Due to this covalent coupling reaction, fluorescent CFSE can be retained within cells for extremely long periods. Also, due to this stable linkage, once incorporated within cells, the dye is not transferred to adjacent cells.
119:
1298:
Hermans IF, Silk JD, Yang J, et al. (February 2004). "The VITAL assay: a versatile fluorometric technique for assessing CTL- and NKT-mediated cytotoxicity against multiple targets in vitro and in vivo".
388:(CFDA-SE), although they are not strictly the same molecule; CFDA-SE, due to its acetate groups, is highly cell permeable, while CFSE is much less so. As CFDA-SE, which is non-fluorescent, enters the
592:
Fazekas de St Groth B, Smith AL, Koh WP, Girgis L, Cook MC, Bertolino P (December 1999). "Carboxyfluorescein diacetate succinimidyl ester and the virgin lymphocyte: a marriage made in heaven".
1426:
Quah BJ, Warren HS, Parish CR (2007). "Monitoring lymphocyte proliferation in vitro and in vivo with the intracellular fluorescent dye carboxyfluorescein diacetate succinimidyl ester".
1015:
Hawkins ED, Hommel M, Turner ML, Battye FL, Markham JF, Hodgkin PD (2007). "Measuring lymphocyte proliferation, survival and differentiation using CFSE time-series data".
338:
385:
1469:
Parish CR, Glidden MH, Quah BJ, Warren HS (February 2009). "Use of the intracellular fluorescent dye CFSE to monitor lymphocyte migration and proliferation".
247:
221:
InChI=1S/C25H15NO9/c27-13-2-5-16-19(10-13)33-20-11-14(28)3-6-17(20)25(16)18-9-12(1-4-15(18)24(32)34-25)23(31)35-26-21(29)7-8-22(26)30/h1-6,9-11,27-28H,7-8H2
231:
InChI=1/C25H15NO9/c27-13-2-5-16-19(10-13)33-20-11-14(28)3-6-17(20)25(16)18-9-12(1-4-15(18)24(32)34-25)23(31)35-26-21(29)7-8-22(26)30/h1-6,9-11,27-28H,7-8H2
1251:"New CFSE-based assay to determine susceptibility to lysis by cytotoxic T cells of leukemic precursor cells within a heterogeneous target cell population"
1058:
Sukkar MB, Stanley AJ, Blake AE, et al. (October 2004). "'Proliferative' and 'synthetic' airway smooth muscle cells are overlapping populations".
522:
Weston SA, Parish CR (October 1990). "New fluorescent dyes for lymphocyte migration studies. Analysis by flow cytometry and fluorescence microscopy".
919:
De Boer RJ, Ganusov VV, MilutinoviΔ D, Hodgkin PD, Perelson AS (July 2006). "Estimating lymphocyte division and death rates from CFSE data".
1130:"High-resolution tracking of cell division suggests similar cell cycle kinetics of hematopoietic stem cells stimulated in vitro and in vivo"
1101:
Khil LY, Kim JY, Yoon JB, et al. (December 1997). "Insulin has a limited effect on the cell cycle progression in 3T3 L1 fibroblasts".
1488:
17:
403:
and track their migration within animals for many months. Subsequent studies revealed that the dye can be used to monitor lymphocyte
212:
1471:
1394:
1377:
428:
176:
637:"Class I-restricted cross-presentation of exogenous self-antigens leads to deletion of autoreactive CD8(+) T cells"
345:
686:"Cell division regulates the T cell cytokine repertoire, revealing a mechanism underlying immune class regulation"
833:
Nordon RE, Nakamura M, Ramirez C, Odell R (December 1999). "Analysis of growth kinetics by division tracking".
1525:
444:
30:
1530:
1129:
16:
This article is about
Carboxyfluorescein succinimidyl ester. For Crystal field stabilization energy, see
1535:
396:, intracellular esterases remove the acetate groups and convert the molecule to the fluorescent ester.
1336:"An in vivo cytotoxicity threshold for influenza A virus-specific effector and memory CD8(+) T cells"
697:
436:
373:
43:
1210:"Tumor-specific CD4+ T cells have a major "post-licensing" role in CTL mediated anti-tumor immunity"
1169:"Flow cytometric analysis of Lactobacillus plantarum to monitor lag times, cell division and injury"
193:
1375:
Lyons AB, Doherty KV (February 2004). "Flow cytometric analysis of cell division by dye dilution".
77:
479:
Parish CR (December 1999). "Fluorescent dyes for lymphocyte migration and proliferation studies".
1502:
1451:
1408:
1280:
1083:
1040:
997:
954:
901:
858:
617:
504:
876:
Gett AV, Hodgkin PD (September 2000). "A cellular calculus for signal integration by T cells".
1494:
1484:
1443:
1400:
1390:
1357:
1316:
1272:
1231:
1190:
1149:
1110:
1075:
1032:
989:
946:
893:
850:
815:
766:
725:
666:
609:
574:
539:
496:
139:
1476:
1435:
1382:
1347:
1308:
1262:
1221:
1180:
1141:
1067:
1024:
981:
936:
928:
885:
842:
805:
797:
756:
715:
705:
656:
648:
601:
566:
531:
488:
456:
270:
94:
1145:
87:
557:
Lyons AB, Parish CR (May 1994). "Determination of lymphocyte division by flow cytometry".
197:
972:
Callard R, Hodgkin P (April 2007). "Modeling T- and B-cell growth and differentiation".
701:
810:
785:
661:
636:
393:
316:
761:
744:
399:
CFSE was originally developed as a fluorescent dye that could be used to stably label
1519:
1185:
1168:
1071:
985:
846:
720:
685:
605:
570:
535:
492:
1412:
1087:
1044:
1001:
164:
1506:
1455:
1284:
958:
905:
862:
621:
508:
786:"B cell differentiation and isotype switching is related to division cycle number"
1480:
1386:
1226:
1209:
1352:
1335:
404:
365:
1267:
1250:
690:
Proceedings of the
National Academy of Sciences of the United States of America
1312:
1249:
Jedema I, van der Werff NM, Barge RM, Willemze R, Falkenburg JH (April 2004).
932:
440:
400:
301:
130:
710:
389:
1498:
1447:
1439:
1404:
1361:
1320:
1276:
1235:
1153:
1079:
1036:
1028:
993:
950:
897:
854:
613:
500:
1194:
1114:
819:
801:
770:
729:
670:
652:
578:
543:
255:
O=C(C5=C4C=C(C(ON6C(CCC6=O)=O)=O)C=C5)OC24C3=C(C=C(O)C=C3)OC1=CC(O)=CC=C12
420:
369:
447:
and even bacteria. Another novel application of CFSE is its use for the
151:
941:
432:
424:
377:
315:
Except where otherwise noted, data are given for materials in their
1167:
Ueckert JE, Nebe von-Caron G, Bos AP, ter Steeg PF (October 1997).
889:
118:
108:
635:
Kurts C, Kosaka H, Carbone FR, Miller JF, Heath WR (July 1997).
745:"Helper T cell differentiation is controlled by the cell cycle"
463:
cells, although remaining viable, proliferate sub-optimally.
1208:
Marzo AL, Kinnear BF, Lake RA, et al. (December 2000).
372:
dye. CFSE is cell permeable and covalently couples, via its
181:
743:
Bird JJ, Brown DR, Mullen AC, et al. (August 1998).
376:, to intracellular molecules, notably, to intracellular
333:
1334:Stambas J, Doherty PC, Turner SJ (February 2007).
1128:Oostendorp RA, Audet J, Eaves CJ (February 2000).
163:
386:carboxyfluorescein diacetate succinimidyl ester
93:
86:
48:2,5-Dioxopyrrolidin-1-yl 3,6-dihydroxy-3-oxo-3
52:-spirobenzofuran-1,9β²-xanthene]-6-carboxylate
8:
784:Hodgkin PD, Lee JH, Lyons AB (July 1996).
196:
138:
22:
1351:
1266:
1225:
1184:
940:
809:
760:
719:
709:
660:
24:6-Carboxyfluorescein succinimidyl ester
471:
252:
217:
192:
1146:10.1182/blood.V95.3.855.003k41_855_862
358:Carboxyfluorescein succinimidyl ester
224:Key: VDABVNMGKGUPEY-UHFFFAOYSA-N
7:
790:The Journal of Experimental Medicine
641:The Journal of Experimental Medicine
684:Gett AV, Hodgkin PD (August 1998).
234:Key: VDABVNMGKGUPEY-UHFFFAOYAV
154:
18:Crystal field stabilization energy
14:
1301:Journal of Immunological Methods
1186:10.1046/j.1472-765x.1997.00225.x
1072:10.1111/j.0818-9641.2004.01275.x
986:10.1111/j.1600-065X.2006.00498.x
921:Bulletin of Mathematical Biology
847:10.1046/j.1440-1711.1999.00869.x
606:10.1046/j.1440-1711.1999.00871.x
559:Journal of Immunological Methods
524:Journal of Immunological Methods
493:10.1046/j.1440-1711.1999.00877.x
323:
288:
282:
29:
1472:Current Protocols in Immunology
1173:Letters in Applied Microbiology
384:CFSE is commonly confused with
319:(at 25 Β°C , 100 kPa).
1378:Current Protocols in Cytometry
291:
276:
1:
762:10.1016/S1074-7613(00)80605-6
1481:10.1002/0471142735.im0409s84
1387:10.1002/0471142956.cy0911s27
1227:10.4049/jimmunol.165.11.6047
571:10.1016/0022-1759(94)90236-4
536:10.1016/0022-1759(90)90322-M
1353:10.4049/jimmunol.178.3.1285
1060:Immunology and Cell Biology
835:Immunology and Cell Biology
594:Immunology and Cell Biology
481:Immunology and Cell Biology
1552:
1268:10.1182/blood-2003-06-2070
15:
1313:10.1016/j.jim.2003.10.017
933:10.1007/s11538-006-9094-8
313:
263:
243:
208:
70:
61:CFSE; Carboxyfluorescein
58:
42:
37:
28:
1381:. Chapter 9: Unit 9.11.
445:hematopoietic stem cells
419:of lymphocytes, such as
711:10.1073/pnas.95.16.9488
1475:. Chapter 4: Unit4.9.
1440:10.1038/nprot.2007.296
1029:10.1038/nprot.2007.297
1340:Journal of Immunology
1214:Journal of Immunology
974:Immunological Reviews
802:10.1084/jem.184.1.277
653:10.1084/jem.186.2.239
457:cytotoxic lymphocytes
44:Preferred IUPAC name
1103:Molecules and Cells
702:1998PNAS...95.9488G
437:smooth muscle cells
309: gΒ·mol
98: (5/6 mixture)
65:-succinimidyl ester
25:
374:succinimidyl group
346:Infobox references
23:
878:Nature Immunology
455:determination of
354:Chemical compound
352:
351:
177:CompTox Dashboard
120:Interactive image
1543:
1511:
1510:
1466:
1460:
1459:
1428:Nature Protocols
1423:
1417:
1416:
1372:
1366:
1365:
1355:
1331:
1325:
1324:
1295:
1289:
1288:
1270:
1246:
1240:
1239:
1229:
1205:
1199:
1198:
1188:
1164:
1158:
1157:
1125:
1119:
1118:
1098:
1092:
1091:
1055:
1049:
1048:
1017:Nature Protocols
1012:
1006:
1005:
969:
963:
962:
944:
916:
910:
909:
873:
867:
866:
830:
824:
823:
813:
781:
775:
774:
764:
740:
734:
733:
723:
713:
681:
675:
674:
664:
632:
626:
625:
589:
583:
582:
554:
548:
547:
519:
513:
512:
476:
336:
330:
327:
326:
308:
293:
290:
284:
278:
271:Chemical formula
201:
200:
185:
183:
167:
156:
142:
122:
97:
90:
33:
26:
1551:
1550:
1546:
1545:
1544:
1542:
1541:
1540:
1516:
1515:
1514:
1491:
1468:
1467:
1463:
1425:
1424:
1420:
1397:
1374:
1373:
1369:
1333:
1332:
1328:
1297:
1296:
1292:
1248:
1247:
1243:
1220:(11): 6047β55.
1207:
1206:
1202:
1166:
1165:
1161:
1127:
1126:
1122:
1100:
1099:
1095:
1057:
1056:
1052:
1014:
1013:
1009:
971:
970:
966:
918:
917:
913:
875:
874:
870:
832:
831:
827:
783:
782:
778:
742:
741:
737:
696:(16): 9488β93.
683:
682:
678:
634:
633:
629:
591:
590:
586:
556:
555:
551:
521:
520:
516:
478:
477:
473:
469:
429:class switching
427:, and antibody
355:
348:
343:
342:
341: ?)
332:
328:
324:
320:
306:
296:
287:
281:
273:
259:
256:
251:
250:
239:
236:
235:
232:
226:
225:
222:
216:
215:
204:
186:
179:
170:
157:
145:
125:
112:
101:
80:
66:
54:
53:
21:
12:
11:
5:
1549:
1547:
1539:
1538:
1533:
1528:
1526:Flow cytometry
1518:
1517:
1513:
1512:
1490:978-0471142737
1489:
1461:
1434:(9): 2049β56.
1418:
1395:
1367:
1346:(3): 1285β92.
1326:
1290:
1261:(7): 2677β82.
1241:
1200:
1159:
1120:
1093:
1050:
1023:(9): 2057β67.
1007:
964:
927:(5): 1011β31.
911:
868:
825:
776:
735:
676:
627:
584:
549:
514:
487:(6): 499β508.
470:
468:
465:
423:production by
353:
350:
349:
344:
322:
321:
317:standard state
314:
311:
310:
304:
298:
297:
294:
285:
279:
274:
269:
266:
265:
261:
260:
258:
257:
254:
246:
245:
244:
241:
240:
238:
237:
233:
230:
229:
227:
223:
220:
219:
211:
210:
209:
206:
205:
203:
202:
194:DTXSID20376335
189:
187:
175:
172:
171:
169:
168:
160:
158:
150:
147:
146:
144:
143:
135:
133:
127:
126:
124:
123:
115:
113:
106:
103:
102:
100:
99:
91:
83:
81:
76:
73:
72:
68:
67:
60:
56:
55:
47:
46:
40:
39:
35:
34:
13:
10:
9:
6:
4:
3:
2:
1548:
1537:
1534:
1532:
1531:Fluorone dyes
1529:
1527:
1524:
1523:
1521:
1508:
1504:
1500:
1496:
1492:
1486:
1482:
1478:
1474:
1473:
1465:
1462:
1457:
1453:
1449:
1445:
1441:
1437:
1433:
1429:
1422:
1419:
1414:
1410:
1406:
1402:
1398:
1392:
1388:
1384:
1380:
1379:
1371:
1368:
1363:
1359:
1354:
1349:
1345:
1341:
1337:
1330:
1327:
1322:
1318:
1314:
1310:
1306:
1302:
1294:
1291:
1286:
1282:
1278:
1274:
1269:
1264:
1260:
1256:
1252:
1245:
1242:
1237:
1233:
1228:
1223:
1219:
1215:
1211:
1204:
1201:
1196:
1192:
1187:
1182:
1178:
1174:
1170:
1163:
1160:
1155:
1151:
1147:
1143:
1140:(3): 855β62.
1139:
1135:
1131:
1124:
1121:
1116:
1112:
1108:
1104:
1097:
1094:
1089:
1085:
1081:
1077:
1073:
1069:
1065:
1061:
1054:
1051:
1046:
1042:
1038:
1034:
1030:
1026:
1022:
1018:
1011:
1008:
1003:
999:
995:
991:
987:
983:
979:
975:
968:
965:
960:
956:
952:
948:
943:
938:
934:
930:
926:
922:
915:
912:
907:
903:
899:
895:
891:
890:10.1038/79782
887:
884:(3): 239β44.
883:
879:
872:
869:
864:
860:
856:
852:
848:
844:
840:
836:
829:
826:
821:
817:
812:
807:
803:
799:
796:(1): 277β81.
795:
791:
787:
780:
777:
772:
768:
763:
758:
755:(2): 229β37.
754:
750:
746:
739:
736:
731:
727:
722:
717:
712:
707:
703:
699:
695:
691:
687:
680:
677:
672:
668:
663:
658:
654:
650:
647:(2): 239β45.
646:
642:
638:
631:
628:
623:
619:
615:
611:
607:
603:
599:
595:
588:
585:
580:
576:
572:
568:
564:
560:
553:
550:
545:
541:
537:
533:
529:
525:
518:
515:
510:
506:
502:
498:
494:
490:
486:
482:
475:
472:
466:
464:
460:
458:
454:
450:
446:
442:
438:
434:
430:
426:
425:T lymphocytes
422:
416:
414:
410:
406:
405:proliferation
402:
397:
395:
391:
387:
382:
379:
375:
371:
367:
363:
359:
347:
340:
335:
318:
312:
305:
303:
300:
299:
275:
272:
268:
267:
262:
253:
249:
242:
228:
218:
214:
207:
199:
195:
191:
190:
188:
178:
174:
173:
166:
162:
161:
159:
153:
149:
148:
141:
137:
136:
134:
132:
129:
128:
121:
117:
116:
114:
110:
105:
104:
96:
92:
89:
85:
84:
82:
79:
75:
74:
69:
64:
57:
51:
45:
41:
36:
32:
27:
19:
1536:Succinimides
1470:
1464:
1431:
1427:
1421:
1376:
1370:
1343:
1339:
1329:
1307:(1): 25β40.
1304:
1300:
1293:
1258:
1254:
1244:
1217:
1213:
1203:
1179:(4): 295β9.
1176:
1172:
1162:
1137:
1133:
1123:
1109:(6): 742β8.
1106:
1102:
1096:
1066:(5): 471β8.
1063:
1059:
1053:
1020:
1016:
1010:
977:
973:
967:
924:
920:
914:
881:
877:
871:
841:(6): 523β9.
838:
834:
828:
793:
789:
779:
752:
748:
738:
693:
689:
679:
644:
640:
630:
600:(6): 530β8.
597:
593:
587:
565:(1): 131β7.
562:
558:
552:
530:(1): 87β97.
527:
523:
517:
484:
480:
474:
461:
452:
448:
417:
412:
408:
398:
383:
361:
357:
356:
71:Identifiers
62:
59:Other names
49:
441:fibroblasts
401:lymphocytes
366:fluorescent
264:Properties
95:117548-22-8
1520:Categories
1396:0471142956
980:: 119β29.
942:1874/22578
467:References
302:Molar mass
131:ChemSpider
107:3D model (
88:92557-81-8
78:CAS Number
390:cytoplasm
1499:19235770
1448:17853860
1413:20204336
1405:18770808
1362:17237374
1321:14871532
1277:14630824
1236:11086036
1154:10648396
1088:25686588
1080:15479432
1045:13550456
1037:17853861
1002:27565949
994:17367338
951:16832737
898:10973282
855:10571673
749:Immunity
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