High-Contrast Luminescent Immunohistochemistry Using PEGylated Lanthanide Complexes.
- Publisher:
- AMER CHEMICAL SOC
- Publication Type:
- Journal Article
- Citation:
- Anal Chem, 2022, 94, (50), pp. 17587-17594
- Issue Date:
- 2022-12-20
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| Filename | Description | Size | |||
|---|---|---|---|---|---|
| acs.analchem.2c04058.pdf | Published version | 7.87 MB |
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Immunohistochemistry (IHC) using fluorescent probes provides high resolution with multiplexing capability, but the imaging contrast is limited by the brightness of the fluorescent probe and the intrinsic autofluorescence background from tissues. Herein, we improved the contrast by high-density labeling of long-lifetime lanthanide complexes and time-gated imaging. As the large (∼280 nm) Stokes shift of lanthanide complexes effectively prevents the issue of concentration quenching, we succeeded in conjugating seven europium complexes to an eight-arm hydrophilic poly(ethylene glycol) (PEG) linker for signal amplification with improved water solubility to the level of up to 10 mg/mL. Moreover, we demonstrated that both human epidermal growth factor receptor 2 (HER2) in a formalin-fixed paraffin-embedded (FFPE) tissue section and cytokeratin 18 (CK18) in a frozen section can be resolved with the enhanced contrast by 2-fold and 3-fold, respectively. Furthermore, we show that the PEGylation of multiple lanthanide complexes is compatible with tyramide signal amplification (TSA). This work suggests new opportunities for sensitive imaging of low-abundance biomarkers in a tissue matrix.
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