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Updated: Jan 7, 2026
Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
Published on: April 8, 2016
In-situ dual-lighting system for HER2 nucleic acid and protein co-localization through simultaneous visual signal
Tiantian Wang1, Zhiqing Chen1, Xuan Fu2
1MOE International Joint Research Laboratory on Synthetic Biology and Medicines, School of Biology and Biological Engineering, South China University of Technology, Guangzhou, China.
Background:
The pathological state of human epidermal growth factor receptor-2 (HER2) determines the treatment strategies. Immunohistochemistry (IHC) and in situ hybridization (ISH) are the only two recognized diagnostic methods for HER2-positive (HER2 + ). IHC, although widely used, has limited reliability and often requires further confirmation by ISH for IHC ≤ 2 + . Furthermore, studies have demonstrated that HER2-low patients may still benefit from HER2-targeted therapy, highlighting the need for accurate and sensitive detection methods.
Methods:
In this study, we designed the molecular beacon (MB) for HER2 amplification detection. Subsequently, the in-situ dual-lighting system (ISDLS) was established by combining MB-based loop-mediated isothermal amplification (LAMP) and tyramide signal amplification (TSA)-based IHC for HER2 in-situ nucleic acid and protein detection in a single sample, and was verified in both cell line slides and human tumor sections. To better elucidate the significance of our study, we compared the test results with clinical reports.
Results:
Here, we show that, in a single sample, satisfactory nucleic acid and protein signals are obtained, especially in human tumor sections within one day. The detection rate for clinical IHC ≥ 1+ and fluorescence in situ hybridization amplification (FISH + ) sections reaches 100%, and the detection rate of clinical FISH- sections reaches 77.8%.
Conclusions:
ISDLS enables the spatial in-situ distribution of amplified nucleic acid and protein signals in one section, providing a basis for multidimensional analysis of diseases. ISDLS is also expected to improve the speed, sensitivity, and accuracy of detection compared to other methods.
