Multiplexed Immunohistochemical Detection of Tumor Markers in Breast Cancer Tissue Using Laser Ablation Inductively Coupled Plasma Mass Spectrometry

Abstract

We optimized multiplexed immunohistochemistry (IHC) on breast cancer tissue. Up to 20 tumor markers are routinely evaluated for one patient, and thus, a common analysis results in a series of time consuming staining procedures. As an alternative, we used lanthanides for labeling of primary antibodies, which are applied in IHC. Laser ablation (LA) ICPMS was elaborated as a detection tool for multiplexed IHC of tissue sections. In this study, we optimized sample preparation steps and LA ICPMS parameters to achieve a sufficient signal-to-background ratio. The results prove the high selectivity of applied antibodies, which was sustained after labeling. Up to three tumor markers (Her 2, CK 7, and MUC 1) were detected simultaneously in a single multiplex analysis of a 5 μm thin breast cancer tissue at a laser spot size of 200 μm. Furthermore, the LA ICPMS results indicate a significantly higher expression level of MUC 1 compared to Her 2 and CK 7, which was not obvious from the conventionally stained tissue sections.

Immunodiagnostics and Immunosensor Design

“…There are efforts to further improve the immunoassays and immunohistochemical tests for CEA [176]. Some further examples are: -carcinoembryonic antigen in lung cancer [177] -tumour markers [178] -gynecological cancer [179] -pancreatic cancer [180] -breast cancer [181,182] Brought to you by | University of Kent Authenticated Download Date | 12/14/17 7:10 PM -breast cancer differential diagnosis [183] -undifferentiated tumours [184], leukemias [185,186], and soft-tissue tumours [187] -renal neoplasms [188] -oral cancer [189] …”

Immunodiagnostics and immunosensor design (IUPAC Technical Report)

“…There are efforts to further improve the immunoassays and immunohistochemical tests for CEA [176]. Some further examples are: -carcinoembryonic antigen in lung cancer [177] -tumour markers [178] -gynecological cancer [179] -pancreatic cancer [180] -breast cancer [181,182] -breast cancer differential diagnosis [183] -undifferentiated tumours [184], leukemias [185,186], and soft-tissue tumours [187] -renal neoplasms [188] -oral cancer [189] …”

Laser ablation ICP-MS for quantitative biomedical applications

“…A further application of LA-ICP-MS in this research area is the use of element-tagged immunoassays for imaging purposes, which provides a new strategy for not only mapping peptides and proteins in different types of tissues, but also for their possible quantification [13,16,88,89]. In this vein, Hutchinson et al [16] reported the imaging of amyloid precursor protein and Aβ peptide in histological sections of a transgenic mouse model of Alzheimer's disease using Eu-and Ni-coupled antibodies.…”

“…The analytical potential of immunohistochemistry combined with LA-ICP-MS has also been demonstrated for the imaging of cancer biomarkers [88,89]. The distributions of two breast cancer-associated proteins (MUC-1 and HER2) were studied based on multiple line scanning of tissue sections by the laser beam and the subsequent measurement of relevant Au/Ag-tagged antibodies bound to the tissue.…”