Single-cell HER2 quantification via instant signal amplification in microdroplets.

Accurate and ultrasensitive evaluation of human epidermal growth factor receptor 2 (HER2) protein is key to early diagnosis and subtype differentiation of breast cancer. Single-cell analyses to reduce ineffective targeted therapies due to breast cancer heterogeneity and improve patient survival remain challenging. Herein, we reported a novel droplet microfluidic combined with an instant cation exchange signal amplification strategy for quantitative analysis of HER2 protein expression on single cells. In the 160 µm droplets produced by a tapered capillary bundle, abundant Immuno-CdS labeled on HER2-positive cells were replaced by Ag + to obtain Cd2+ that stimulated Rhod-5N fluorescence. This uniformly distributed and instantaneous fluorescence amplification strategy in droplets improves sensitivity and reduces signal fluctuation. Using HER2 modified PS microsphere to simulate single cells, we obtained a linear fitting of HER2-modified concentration and fluorescence intensity in microdroplets with the limit detection of 11.372 pg mL-1. Moreover, the relative standard deviation (RSD) was 4.2-fold lower than the traditional immunofluorescence technique (2.89% vs 12.21%). The HER2 protein on SK-BR-3 cells encapsulated in droplets was subsequently quantified, ranging from 9862.954 pg mL-1 and 205.26 pg mL-1, equivalent to 9.795 × 106 and 2.038 × 105 protein molecules. This detection system provides a universal platform for single-cell sensitive quantitative analysis and contributes to the evaluation of HER2-positive tumors.

All-fiber online Raman sensor with enhancement via a Fabry-Perot cavity.

In this Letter, a novel all-fiber online Raman sensor with significant signal enhancement via a Fabry-Perot (FP) cavity is proposed and demonstrated. The FP cavity structure is formed by inserting a long-pass coated fiber and a gold-plated capillary into a silver-lined capillary with a gap. A corroded single-mode fiber is inserted into the gold-plated capillary to guide the excitation light into the FP cavity. The multiple reflections of excitation light in the FP cavity have significantly increased the interaction volume between the light and the sample. Experiment results have demonstrated an enhancement factor of 5 times in the detected Raman signal for ethanol compared to that measured using the silver-lined hollow-core fiber-based Raman cell without FP cavity, or 86 times compared with direct detection using a bare fiber tip. The measurement results are in good agreement with theoretical analyses. This Raman sensor with signal enhancement via the FP cavity has the potential to realize rapid sample replacement and online detection with high sensitivity and high accuracy for biochemical applications.

Volumetric enhancement of Raman scattering for fast detection based on a silver-lined hollow-core fiber.

Fast detection and identification of chemicals are of utmost importance for field testing and real-time monitoring in many fields. Raman spectroscopy is the predominant technique in principle, but its wide application is limited on account of weak scattering efficiency. Surface Enhanced Raman Spectroscopy (SERS) technique provides a solution for signal enhancement, but may not good at fast detection due to cross contamination and bulky instruments. Hollow-core fiber-based Raman cell with long interaction length can achieve high detection sensitivity, but it also suffers from low flow rate, bulky high-pressure equipment and light coupling structure, which also restricts its application for fast detection. In order to solve those problems, we proposed a portable Raman cell, by using metal-lined hollow-core fibers (MLHCF) with large bandwidth, good field confinement, extremely large numerical aperture and arbitrary length. With our proposed fiber inserted light coupling and light reflecting method, a Raman cell of 3.1 cm in length provides nearly 50 times of signal enhancement compared with direct detection using bare fiber tip. Furthermore, the sample exchange rate could be as fast as 1 second even under normal pressure without any cross contamination. At last, we also demonstrated the underlying general mechanism of signal enhancement and summarized it as volumetric enhancement of Raman scattering (VERS). Both the experiment results and the theoretical analysis demonstrated that our device has the potential for fast online Raman detection, which also possesses high-sensitivity and high-accuracy.