A guide to integrating immunohistochemistry and chemical imaging

Abstract

Chemical imaging provides new insight into the fundamental atomic, molecular, and biochemical composition of tissue and how they are interrelated in normal physiology. Visualising and quantifying products of pathogenic reactions long before structural changes become apparent also adds a new dimension to understanding disease pathogenesis. While chemical imaging in isolation is somewhat limited by the nature of information it can provide (e.g. peptides, metals, lipids, or functional groups), integrating immunohistochemistry allows simultaneous, targeted imaging of biomolecules while also mapping tissue composition. Together, this approach can provide invaluable information on the inner workings of the cell and the molecular basis of diseases. Key learning points(1) Chemical imaging must be redefined to encompass all analytical techniques that can spatially map the chemical composition of a sample.

Nanocatalytic Medicine

“…In addition to the aforementioned clinically used imaging modalities, histochemistry has also been extensively investigated as a diagnostic tool for visually assessing tissue morphology and underlying pathology . Histochemical assay takes advantage of one or more disease‐associated endogenous biochemicals that can react with certain dyes to form colored compounds, thus enabling the visualization of anatomical pathology based on the distribution of these pathological chemicals.…”

Quantitative mapping of specific proteins in biological tissues by laser ablation–ICP-MS using exogenous labels: aspects to be considered

“…There is no doubt that the combination of IHC with spectroscopic and MS techniques offers great possibilities for quantitative imaging of specific proteins in biological tissues, with detailed aid in experimental design provided in previous reviews [3]. Finally, it is worth to mention that research in this field can be extended to other cutting edge studies, such as the elucidation of mechanisms related with antibody-drug conjugates for oncology therapeutics [67,68], as well as for nucleic acids (DNA and RNA) assays [69].…”

“…In principle, such protocols can be similar than those used in conventional immunofluorescence and immunoperoxidase staining, excepting that the label (chemical reporter) should contain an element (or isotope) detectable by ICP-MS. In addition, special care should be taken when using conventional IHC protocols, because the microchemical composition can be altered during the sample preparation procedure (e.g., formalin fixation and paraffin embedding) [3,37]. surfactant is added to the slides.…”

“…Each of both alternatives has inherent advantages: labelled primary antibodies offer higher specificity and greater potential for multiplexing, while the use of labelled secondary antibodies allows for higher sensitivity and may be used as generic platform [3].…”

“…the capability to assess not only differential protein expression between two samples, but also protein absolute quantities) [1,2]. Optical-spectroscopy and mass spectrometry (MS) techniques offer interesting capabilities within this field for liquid samples and for chemical imaging of solid specimens [3,4].…”