Label-Free Detection of Insulin and Glucagon within Human Islets of Langerhans Using Raman Spectroscopy

J. Hilderink, Cornelis Otto, Cornelis H. Slump, Aufrid T.M. Lenferink, M.A. Engelse, Clemens van Blitterswijk, E.J.P. de Koning, Hermanus Bernardus Johannes Karperien, Aart A. van Apeldoorn

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Abstract

Intrahepatic transplantation of donor islets of Langerhans is a promising therapy for patients with type 1 diabetes. It is of critical importance to accurately monitor islet quality before transplantation, which is currently done by standard histological methods that are performed off-line and require extensive sample preparation. As an alternative, we propose Raman spectroscopy which is a non-destructive and label-free technique that allows continuous real-time monitoring of the tissue to study biological changes as they occur. By performing Raman spectroscopic measurements on purified insulin and glucagon, we showed that the 520 cm-1 band assigned to disulfide bridges in insulin, and the 1552 cm-1 band assigned to tryptophan in glucagon are mutually exclusive and could therefore be used as indirect markers for the label-free distinction between both hormones. High-resolution hyperspectral Raman imaging for these bands showed the distribution of disulfide bridges and tryptophan at sub-micrometer scale, which correlated with the location of insulin and glucagon as revealed by conventional immunohistochemistry. As a measure for this correlation, quantitative analysis was performed comparing the Raman images with the fluorescence images, resulting in Dice coefficients (ranging between 0 and 1) of 0.36 for insulin and 0.19 for glucagon. Although the use of separate microscope systems with different spatial resolution and the use of indirect Raman markers cause some image mismatch, our findings indicate that Raman bands for disulfide bridges and tryptophan can be used as distinctive markers for the label-free detection of insulin and glucagon in human islets of Langerhans.
Original languageEnglish
Pages (from-to)e78148-e78148
JournalPLoS ONE
Volume8
Issue number10
DOIs
Publication statusPublished - 22 Oct 2013

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Raman spectroscopy
Raman Spectrum Analysis
glucagon
islets of Langerhans
Glucagon
Islets of Langerhans
Labels
insulin
Insulin
sulfides
Tryptophan
Disulfides
tryptophan
Islets of Langerhans Transplantation
insulin-dependent diabetes mellitus
monitoring
Medical problems
Type 1 Diabetes Mellitus
microscopes
immunohistochemistry

Keywords

  • IR-87563
  • METIS-298330

Cite this

Hilderink, J. ; Otto, Cornelis ; Slump, Cornelis H. ; Lenferink, Aufrid T.M. ; Engelse, M.A. ; van Blitterswijk, Clemens ; de Koning, E.J.P. ; Karperien, Hermanus Bernardus Johannes ; van Apeldoorn, Aart A. / Label-Free Detection of Insulin and Glucagon within Human Islets of Langerhans Using Raman Spectroscopy. In: PLoS ONE. 2013 ; Vol. 8, No. 10. pp. e78148-e78148.
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Label-Free Detection of Insulin and Glucagon within Human Islets of Langerhans Using Raman Spectroscopy. / Hilderink, J.; Otto, Cornelis; Slump, Cornelis H.; Lenferink, Aufrid T.M.; Engelse, M.A.; van Blitterswijk, Clemens; de Koning, E.J.P.; Karperien, Hermanus Bernardus Johannes; van Apeldoorn, Aart A.

In: PLoS ONE, Vol. 8, No. 10, 22.10.2013, p. e78148-e78148.

Research output: Contribution to journalArticleAcademicpeer-review

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AU - Hilderink, J.

AU - Otto, Cornelis

AU - Slump, Cornelis H.

AU - Lenferink, Aufrid T.M.

AU - Engelse, M.A.

AU - van Blitterswijk, Clemens

AU - de Koning, E.J.P.

AU - Karperien, Hermanus Bernardus Johannes

AU - van Apeldoorn, Aart A.

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N2 - Intrahepatic transplantation of donor islets of Langerhans is a promising therapy for patients with type 1 diabetes. It is of critical importance to accurately monitor islet quality before transplantation, which is currently done by standard histological methods that are performed off-line and require extensive sample preparation. As an alternative, we propose Raman spectroscopy which is a non-destructive and label-free technique that allows continuous real-time monitoring of the tissue to study biological changes as they occur. By performing Raman spectroscopic measurements on purified insulin and glucagon, we showed that the 520 cm-1 band assigned to disulfide bridges in insulin, and the 1552 cm-1 band assigned to tryptophan in glucagon are mutually exclusive and could therefore be used as indirect markers for the label-free distinction between both hormones. High-resolution hyperspectral Raman imaging for these bands showed the distribution of disulfide bridges and tryptophan at sub-micrometer scale, which correlated with the location of insulin and glucagon as revealed by conventional immunohistochemistry. As a measure for this correlation, quantitative analysis was performed comparing the Raman images with the fluorescence images, resulting in Dice coefficients (ranging between 0 and 1) of 0.36 for insulin and 0.19 for glucagon. Although the use of separate microscope systems with different spatial resolution and the use of indirect Raman markers cause some image mismatch, our findings indicate that Raman bands for disulfide bridges and tryptophan can be used as distinctive markers for the label-free detection of insulin and glucagon in human islets of Langerhans.

AB - Intrahepatic transplantation of donor islets of Langerhans is a promising therapy for patients with type 1 diabetes. It is of critical importance to accurately monitor islet quality before transplantation, which is currently done by standard histological methods that are performed off-line and require extensive sample preparation. As an alternative, we propose Raman spectroscopy which is a non-destructive and label-free technique that allows continuous real-time monitoring of the tissue to study biological changes as they occur. By performing Raman spectroscopic measurements on purified insulin and glucagon, we showed that the 520 cm-1 band assigned to disulfide bridges in insulin, and the 1552 cm-1 band assigned to tryptophan in glucagon are mutually exclusive and could therefore be used as indirect markers for the label-free distinction between both hormones. High-resolution hyperspectral Raman imaging for these bands showed the distribution of disulfide bridges and tryptophan at sub-micrometer scale, which correlated with the location of insulin and glucagon as revealed by conventional immunohistochemistry. As a measure for this correlation, quantitative analysis was performed comparing the Raman images with the fluorescence images, resulting in Dice coefficients (ranging between 0 and 1) of 0.36 for insulin and 0.19 for glucagon. Although the use of separate microscope systems with different spatial resolution and the use of indirect Raman markers cause some image mismatch, our findings indicate that Raman bands for disulfide bridges and tryptophan can be used as distinctive markers for the label-free detection of insulin and glucagon in human islets of Langerhans.

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KW - METIS-298330

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M3 - Article

VL - 8

SP - e78148-e78148

JO - PLoS ONE

JF - PLoS ONE

SN - 1932-6203

IS - 10

ER -