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Label-free fluorescence detection in capillary and microchip electrophoresis

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Abstract

Herein, we summarize the current status of native fluorescence detection in microchannel electrophoresis, with a strong focus on chip-based systems. Fluorescence detection is a powerful technique with unsurpassed sensitivity down to the single-molecule level. Accordingly fluorescence detection is attractive in combination with miniaturised separation techniques. A drawback is, however, the need to derivatize most analytes prior to analysis. This can often be circumvented by utilising excitation light in the UV spectral range in order to excite intrinsic fluorescence. As sensitive absorbance detection is challenging in chip-based systems, deep-UV fluorescence detection is currently one of the most general optical detection techniques in microchip electrophoresis, which is especially attractive for the detection of unlabelled proteins. This review gives an overview of research on native fluorescence detection in capillary (CE) and microchip electrophoresis (MCE) between 1998 and 2008. It discusses material aspects of native fluorescence detection and the instrumentation used, with particular focus on the detector design. Newer developments, featured techniques, and their prospects in the future are also included. In the last section, applications in bioanalysis, drug determination, and environmental analysis are reviewed with regard to limits of detection.

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Abbreviations

CE:

Capillary electrophoresis

MCE:

Microchip electrophoresis

HPLC:

High-performance liquid chromatography

LIF:

Laser-induced fluorescence

MS:

Mass spectrometry

trp:

Tryptophan

LED:

Light-emitting diode

fq:

Frequency-quadrupled

fd:

Frequency-doubled

NdYAG:

Neodymium-doped yttrium aluminium garnet

FS:

Fused silica

COC:

Cyclic olefin copolymer

PP:

Polypropylene

PE:

Polyethylene

PDMS:

Polydimethylsiloxane

NA:

Numerical aperture

PMT:

Photomultiplier tube

APD:

Avalanche photodiode

OPE:

One-photon excitation

MPE:

Multiphoton excitation

PAH:

Polycyclic aromatic hydrocarbons

References

  1. Zubritsky E (2002) Anal Chem 74:22A–26A

    CAS  Google Scholar 

  2. Dolnik V (2008) Electrophoresis 29:143–156

    Article  CAS  Google Scholar 

  3. Kleparnik K, Bocek P (2007) Chem Rev 107:5279–5317

    Article  CAS  Google Scholar 

  4. Sinville R, Soper SA (2007) J Sep Sci 30:1714–1728

    Article  CAS  Google Scholar 

  5. Horsman KM, Bienvenue JM, Blasier KR, Rainer M, Bonn GK (2007) J Forensic Sci 52:784–799

    Article  CAS  Google Scholar 

  6. Manz A, Harrison DJ, Verpoorte EMJ, Fettinger JC, Paulus A, Ludi H, Widmer HM (1992) J Chromatogr 593:253–258

    Article  CAS  Google Scholar 

  7. Ludwig M, Belder D, Wang LW, Reetz MT (2006) Angew Chem Int Ed 45:2463–2466

    Article  Google Scholar 

  8. Breadmore MC (2007) Electrophoresis 28:254–281

    Article  CAS  Google Scholar 

  9. Götz S, Karst U (2007) Anal Bioanal Chem 387:183–192

    Article  Google Scholar 

  10. Dittrich PS, Manz A (2005) Anal Bioanal Chem 382:1771–1782

    Article  CAS  Google Scholar 

  11. Kuswandi B, Nuriman, Huskens J, Verboom W (2007) Anal Chim Acta 601:141–155

    Article  CAS  Google Scholar 

  12. Matysik FM (2008) Microchim Acta 160:1–14

    Article  CAS  Google Scholar 

  13. Du Y, Wang EK (2007) J Sep Sci 30:875–890

    Article  CAS  Google Scholar 

  14. Lazar LM, Grym J, Foret F (2005) Mass Spectrom Rev 25:573–594

    Article  Google Scholar 

  15. Wu YT, Chen YC (2005) Anal Chem 77:2071–2077

    Article  CAS  Google Scholar 

  16. Klampfl CW (2006) Electrophoresis 27:3–34

    Article  CAS  Google Scholar 

  17. Campa C, Coslovi A, Flamigni A, Rossi M (2006) Electrophoresis 27:2027–2050

    Article  CAS  Google Scholar 

  18. Viskari PJ, Landers JP (2006) Electrophoresis 27:1797–1810

    Article  CAS  Google Scholar 

  19. Xue Q, Foret F, Dunayevskiy YM, Zavracky PM, McGruer NE, Karger BL (1997) Anal Chem 69:426–430

    Article  CAS  Google Scholar 

  20. Herrero M, Ibanez E, Cifuentes A (2007) Electrophoresis 28:2148–2160

    Article  Google Scholar 

  21. Ramsey RS, Ramsey JM (1997) Anal Chem 69:1174–1178

    Article  CAS  Google Scholar 

  22. Foret F, Kusy P (2006) Electrophoresis 27:4877–4887

    Article  CAS  Google Scholar 

  23. Hoffmann P, Häusig U, Schulze P, Belder D (2007) Angew Chem Int Ed 46:4913–4916

    Article  CAS  Google Scholar 

  24. Kelly RT, Tang K, Irimia D, Toner M, Smith RD (2008) Anal Chem 80:3824–3831

    Article  CAS  Google Scholar 

  25. Zamfir AD, Blindila L, Lion N, Allen M, Girault HH, Peter-Katalinic J (2005) Electrophoresis 26:3650–3673

    Article  CAS  Google Scholar 

  26. Kuban P, Hauser PC (2008) Anal Chim Acta 607:15–29

    Article  CAS  Google Scholar 

  27. Pumera M (2007) Talanta 74:358–364

    Article  CAS  Google Scholar 

  28. Bendrysheva SN, Proskurnin MA, Pyell U, Faubel W (2006) Anal Bioanal Chem 385:1492–1503

    Article  CAS  Google Scholar 

  29. Yamauchi M, Mawatari K, Hibara A, Tokeshi M, Kitamori T (2006) Anal Chem 78:2646–2650

    Article  CAS  Google Scholar 

  30. Tamaki E, Hibara A, Tokeshi M, Kitamori T (2004) Lab Chip 5:129–131

    Article  Google Scholar 

  31. Hiki S, Mawatari K, Hibara A, Tokeshi M, Kitamori T (2006) Anal Chem 78:2859–2863

    Article  CAS  Google Scholar 

  32. Gooijer C, Kok SJ, Ariese F (2000) Analusis 28:679–685

    Article  CAS  Google Scholar 

  33. Lacroix M, Poinsot V, Fournier C, Couderc F (2005) Electrophoresis 26:2608–2621

    Article  CAS  Google Scholar 

  34. Johnson ME, Landers JP (2004) Electrophoresis 25:3513–3527

    Article  CAS  Google Scholar 

  35. Wabuyele MB, Ford SM, Stryjewski W, Barrow J, Soper SA (2004) Electrophoresis 22:3939–3948

    Article  Google Scholar 

  36. Shear JB (1999) Anal Chem 71:598A–605A

    CAS  Google Scholar 

  37. Jameson DM, Weber G (1981) J Phys Chem 85:953–958

    Article  CAS  Google Scholar 

  38. Parpura V, Tong W, Yeung ES, Haydon PG (1998) J Neurosci Methods 82:151–158

    Article  CAS  Google Scholar 

  39. Kok SJ, Hoornweg GP, de Ridder T, Brinkman UAT, Velthorst NH, Gooijer C (1998) J Chromatogr A 806:355–360

    Article  CAS  Google Scholar 

  40. Kok SJ, Isberg ICK, Gooijer C, Brinkman UAT Velthorst NH (1998) Anal Chim Acta 360:109–118

    Article  CAS  Google Scholar 

  41. Park YH, Zhang X, Rubakhin S, Sweedler JV (1999) Anal Chem 71:4997–5002

    Article  CAS  Google Scholar 

  42. Horstkötter C, Blaschke G (2001) J Chromatogr B 754:169–178

    Article  Google Scholar 

  43. Hellmich W, Pelargus C, Leffhalm K, Ros A, Anselmetti D (2005) Electrophoresis 26:3689–3696

    Article  CAS  Google Scholar 

  44. Schulze P, Ludwig M, Kohler F, Belder D (2005) Anal Chem 77:1325–1329

    Article  CAS  Google Scholar 

  45. Lapainis T, Scanlan C, Rubakhin SS, Sweedler JV (2007) Anal Bioanal Chem 387:97–105

    Article  CAS  Google Scholar 

  46. Sluszny C, He Y, Yeung ES (2005) Electrophoresis 26:4197–4203

    Article  CAS  Google Scholar 

  47. Lee J, Tripathi A (2007) Biotechnol Prog 23:1506–1512

    Article  CAS  Google Scholar 

  48. Taniyasu Y, Kasu M, Makimoto T (2006) Nature 441:325–328

    Article  CAS  Google Scholar 

  49. Burgess DS (2006) Photonics Spectra 40:119–119

    Google Scholar 

  50. Williams DC, Soper SA (1995) Anal Chem 67:3427–3432

    Article  CAS  Google Scholar 

  51. Timperman AT, Khatib K, Sweedler JV (1995) Anal Chem 67:139–144

    Article  CAS  Google Scholar 

  52. Lakowicz JR (2006) Principles of fluorescence spectroscopy. Springer, Berlin Heidelberg New York

    Google Scholar 

  53. Shan JW, Lang DB, Dimotakis PE (2004) Exp Fluids 36:268–273

    Article  CAS  Google Scholar 

  54. Schulze P, Ludwig M, Belder D, Electrophoresis, in press

  55. Hellmich W, Greif D, Pelargus C, Anselmetti D, Ros A (2006) J Chromatogr A 1130:195–200

    Article  CAS  Google Scholar 

  56. Timpermann AT, Oldenburg KE, Sweedler JV (1995) Anal Chem 67:3421–3426

    Article  Google Scholar 

  57. Denk W, Strickler JH, Webb WW (1990) Science 248:73–76

    Article  CAS  Google Scholar 

  58. Zugel SA, Burke BJ, Regnier FE, Lytle FE (2000) Anal Chem 72:5731–5735

    Article  CAS  Google Scholar 

  59. Birch DJS, McLoskey D, Sanderson A, Suhling K, Holmes AS (1994) J Fluoresc 4:91–102

    Article  CAS  Google Scholar 

  60. Okerberg E, Shear JB (2001) Anal Biochem 292:311–313

    Article  CAS  Google Scholar 

  61. Wise DD, Shear JB (2006) J Chromatogr A 1111:153–158

    Article  CAS  Google Scholar 

  62. Paul UP, Li L, Lee ML, Farnsworth PB (2005) Anal Chem 77:3690–3693

    Article  CAS  Google Scholar 

  63. Schulze P, Schüttpelz M, Sauer M, Belder D (2007) Lab Chip 7:1841–1844

    Article  CAS  Google Scholar 

  64. Yeung ES (1999) J Chromatogr A 830:243–262

    Article  CAS  Google Scholar 

  65. Cannon DM, Winograd N, Ewing AG (2000) Annu Rev Biophys Biomol Struct 29:239–263

    Article  CAS  Google Scholar 

  66. Tagliaro F, Bortolotti F (2008) Electrophoresis 29:260–268

    Article  CAS  Google Scholar 

  67. Chiu TC, Lin YW, Huang YF, Chang HT (2006) Electrophoresis 27:4792–4807

    Article  CAS  Google Scholar 

  68. Couderc F, Causse E, Bayle C (1998) Electrophoresis 19:2777–2790

    Article  CAS  Google Scholar 

  69. Chan C, Muschik GM, Issaq HJ (2000) Electrophoresis 21:2062–2066

    Article  CAS  Google Scholar 

  70. Miao H, Rubakhin SS, Sweedler JV (2003) Anal Bioanal Chem 377:1007–1013

    Article  CAS  Google Scholar 

  71. Fuller RR, Moroz LL, Gilette R, Sweedler JV (1998) Neuron 20:173–181

    Article  CAS  Google Scholar 

  72. Squires LN, Talbot KN, Rubakhin SS, Sweedler JV (2007) J Neurochem 103:174–180

    CAS  Google Scholar 

  73. Squires LN, Jakubowski JA, Stuart JN, Rubakhin SS, Hatcher NG, Kim WS, Chen K, Shih JC, Seif I, Sweedler JV (2006) J Biol Chem 281:13463–13470

    Article  CAS  Google Scholar 

  74. Beekman MC, Lingeman H, Brinkman AUT Gooijer C (1999) J Microcolumn Sep 11:347–352

    Article  CAS  Google Scholar 

  75. Kuijt J, Garcia-Ruiz C, Stroomberg GJ, Marina ML, Ariese F, Brinkmann UAT, Gooijer C (2001) J Chromatogr A 907:291–299

    Article  CAS  Google Scholar 

  76. Tolba K, Belder D (2007) Electrophoresis 28:2934–2941

    Article  CAS  Google Scholar 

  77. Zaugg S, Zhang X, Sweedler J, Thormann W (2001) J Chromatogr B 752:17–31

    Article  CAS  Google Scholar 

  78. Huhn C, Pütz M, Martin N, Dahlenburg R, Pyell U (2005) Electrophoresis 26:2391–2401

    Article  CAS  Google Scholar 

  79. Soetebeer UB, Schierenberg MO, Schulz H, Grünefeld G, Andresen P, Blaschke G (2000) J Chromatogr B 745:271–278

    Article  CAS  Google Scholar 

  80. Alnajjar A, Butcher JA, McCord B (2004) Electrophoresis 25:1592–1600

    Article  CAS  Google Scholar 

  81. Plenert ML, Shear JB (2003) Proc Natl Acad Sci USA 100:3853–3857

    Article  CAS  Google Scholar 

  82. Burns MA, Johnson BN, Brahmasandra SN, Handique K, Webster JR, Krishnan M, Sammarco TS, Man PM, Jones D, Heldsinger D, Mastrangelo CH, Burke DT (1998) Science 282:484–487

    Article  CAS  Google Scholar 

  83. Yao B, Luo G, Wang LD, Gao YD, Lei GT, Ren KN, Chen LX, Wang YM, Hu Y, Qiu Y (2005) Lab Chip 5:1041–1047

    Article  Google Scholar 

  84. Miyaki K, Guo YL, Shimosaka T, Nakagama T, Nakajima H, Uchiyama K (2005) Anal Bioanal Chem 382:810–816

    Article  CAS  Google Scholar 

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  1. Institute of Analytical Chemistry, Universitat Leipzig, Johannisallee 29, 04103, Leipzig, Germany

    Philipp Schulze & Detlev Belder

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  1. Philipp Schulze
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  2. Detlev Belder
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Correspondence to Detlev Belder.

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Schulze, P., Belder, D. Label-free fluorescence detection in capillary and microchip electrophoresis. Anal Bioanal Chem 393, 515–525 (2009). https://doi.org/10.1007/s00216-008-2452-7

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