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Updated: Mar 15, 2026
A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections
Published on: March 30, 2020
Multiplexed Molecular Imaging of Fresh Tissue Surfaces Enabled by Convection-Enhanced Topical Staining with
Yu W Wang1, Josh D Doerksen2, Soyoung Kang2
1Department of Mechanical Engineering, University of Washington, Seattle, WA, 98195, USA. yuwang2@uw.edu.
Abstract:
There is a need for intraoperative imaging technologies to guide breast-conserving surgeries and to reduce the high rates of re-excision for patients in which residual tumor is found at the surgical margins during postoperative pathology analyses. Feasibility studies have shown that utilizing topically applied surface-enhanced Raman scattering (SERS) nanoparticles (NPs), in conjunction with the ratiometric imaging of targeted versus untargeted NPs, enables the rapid visualization of multiple cell-surface biomarkers of cancer that are overexpressed at the surfaces of freshly excised breast tissues. In order to reliably and rapidly perform multiplexed Raman-encoded molecular imaging of large numbers of biomarkers (with five or more NP flavors), an enhanced staining method has been developed in which tissue surfaces are cyclically dipped into an NP-staining solution and subjected to high-frequency mechanical vibration. This dipping and mechanical vibration (DMV) method promotes the convection of the SERS NPs at fresh tissue surfaces, which accelerates their binding to their respective biomarker targets. By utilizing a custom-developed device for automated DMV staining, this study demonstrates the ability to simultaneously image four cell-surface biomarkers of cancer at the surfaces of fresh human breast tissues with a mixture of five flavors of SERS NPs (four targeted and one untargeted control) topically applied for 5 min and imaged at a spatial resolution of 0.5 mm and a raster-scanned imaging rate of >5 cm2 min-1 .
Insights
A new dipping and mechanical vibration (DMV) method enhances surface-enhanced Raman scattering (SERS) nanoparticle (NP) staining for intraoperative breast cancer imaging. This technique rapidly visualizes multiple cancer biomarkers on excised tissue, improving surgical accuracy.
Area of Science:
- Biomedical Engineering
- Molecular Imaging
- Surgical Oncology
Background:
- Intraoperative imaging is crucial for guiding breast-conserving surgeries and minimizing re-excision rates due to residual tumor.
- Surface-enhanced Raman scattering (SERS) nanoparticles (NPs) show promise for visualizing cancer biomarkers on excised breast tissue.
- Multiplexed imaging of multiple biomarkers requires efficient and rapid NP staining methods.
Purpose of the Study:
- To develop an enhanced staining method for rapid, multiplexed Raman-encoded molecular imaging of breast cancer biomarkers.
- To evaluate the efficacy of a novel dipping and mechanical vibration (DMV) technique for SERS NP application.
- To demonstrate simultaneous imaging of multiple cell-surface cancer biomarkers on fresh human breast tissues.
Main Methods:
- Developed a dipping and mechanical vibration (DMV) method for cyclic NP staining of tissue surfaces.
- Utilized a mixture of five SERS NP flavors (four targeted, one control) for multiplexed imaging.
- Employed a custom-built device for automated DMV staining and high-frequency mechanical vibration.
- Performed imaging at a spatial resolution of 0.5 mm with a raster-scanned imaging rate exceeding 5 cm²/min.
Main Results:
- The DMV method significantly enhances SERS NP convection and binding to target biomarkers on fresh tissue surfaces.
- Successfully demonstrated simultaneous imaging of four distinct cell-surface cancer biomarkers.
- Achieved rapid topical application of SERS NPs within 5 minutes for imaging.
- Validated the feasibility of multiplexed molecular imaging for intraoperative guidance.
Conclusions:
- The automated DMV staining method enables rapid and reliable multiplexed Raman-encoded molecular imaging of breast cancer biomarkers.
- This technology has the potential to improve intraoperative guidance during breast-conserving surgeries, reducing re-excision rates.
- The developed technique offers a significant advancement in molecular imaging for surgical pathology.
