Multiplex bioimaging of proteins-related to neurodegenerative diseases in eye sections by laser ablation - Inductively coupled plasma – Mass spectrometry using metal nanoclusters as labels

Ana Lores-Padín, Beatriz Fernández*, Lydia Álvarez, Héctor González-Iglesias, Imre Lengyel, Rosario Pereiro

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)
11 Downloads (Pure)

Abstract

Simultaneous determination of proteins with micrometric resolution is a significant challenge. In this study, laser ablation (LA) inductively coupled plasma - mass spectrometry (ICP-MS) was employed to quantify the distribution of proteins associated to the eye disease age-related macular degeneration (AMD) using antibodies labelled with three different metal nanoclusters (MNCs). PtNCs, AuNCs and AgNCs contain hundreds of metal atoms and were used to detect metallothionein 1/2 (MT1/2), complement factor H (CFH) and amyloid precursor protein (APP) in retina, ciliary body, retinal pigment epithelium (RPE), choroid and sclera from human cadaveric eye sections. First, the labelling of MNCs bioconjugated primary antibodies (Ab) was optimised following an immunolabelling protocol to avoid the non-specific interaction of MNCs with the tissue. Then, the LA and ICP-MS conditions were studied to obtain high-resolution images for the simultaneous detection of the three labels at the same tissue section. A significant signal amplification was found when using AuNCs, AgNCs and PtNCs labelled Ab of 310, 723 and 1194 respectively. After the characterisation of MNCs labelled immunoprobes, the Ab labelling was used for determination of MT1/2, CFH and APP in the RPE-choroid-sclera, where accumulation of extracellular deposits related to AMD was observed. Experimental results suggest that this method is fully suitable for the simultaneous detection of at least three different proteins.

Original languageEnglish
Article number121489
Number of pages10
JournalTalanta
Volume221
Early online date05 Aug 2020
DOIs
Publication statusPublished - 01 Jan 2021

Keywords

  • Age-related macular degeneration
  • Bioimaging
  • Immunohistochemistry
  • Laser ablation ICP-MS
  • Metal nanoclusters
  • Multiplex analysis of proteins

ASJC Scopus subject areas

  • Analytical Chemistry

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