Dual-Mode Undistorted Visual Fluorescent Sensing Strategy through Manipulating the Coffee-Ring Effect on Microfluidic Paper-Based Chip.

To overcome the insufficient sensitivity due to distortion of the fluorescent images by mobile devices, we first developed a novel dual-mode strategy for undistorted visual fluorescent sensing on µPAD by technically manipulating the coffee-ring effect of the fluid sample. Based on the manipulating coffee-ring effect, we divided the horizontal direction of the resulting fluorescence image into 600 pixels and obtained more accurate quantitative information to avoid image distortion. The bovine serum albumin-stabilized gold nanoclusters-copper ion complex was used as the fluorescent probe, combined with a small imaging box and a smartphone, to achieve a rapid testing of histidine in human urine. The output image was analyzed in dual mode: RGB numerical analysis in pixel units and the direct measurement of the fluorescent strips length (limit of detection (LOD) is 0.021 and 0.5 mM, respectively), and improved antidistortion for visual fluorescent sensing. This strategy can overcome the distortion of a smartphone-visualized fluorescent image and shows great potential for rapid and convenient analysis.

Occurrence, Distribution, and Trophic Transfer of Pharmaceuticals and Personal Care Products in the Bohai Sea.

The ubiquitous presence of pharmaceuticals and personal care products (PPCPs) in environments has aroused global concerns; however, minimal information is available regarding their multimedia distribution, bioaccumulation, and trophic transfer in marine environments. Herein, we analyzed 77 representative PPCPs in samples of surface and bottom seawater, surface sediments, and benthic biota from the Bohai Sea. PPCPs were pervasively detected in seawater, sediments, and benthic biota, with antioxidants being the most abundant PPCPs. PPCP concentrations positively correlated between the surface and bottom water with a decreasing trend from the coast to the central oceans. Higher PPCP concentrations in sediment were found in the Yellow River estuary, and the variations in the physicochemical properties of PPCPs and sediment produced a different distribution pattern of PPCPs in sediment from seawater. The log Dow, but not log Kow, showed a linear and positive relationship with bioaccumulation and trophic magnification factors and a parabolic relationship with biota-sediment accumulation factors. The trophodynamics of miconazole and acetophenone are reported for the first time. This study provides novel insights into the multimedia distribution and biomagnification potential of PPCPs and suggests that log Dow is a better indicator of their bioaccumulation and trophic magnification.

In Situ Sea Cucumber Detection across Multiple Underwater Scenes Based on Convolutional Neural Networks and Image Enhancements.

Recently, rapidly developing artificial intelligence and computer vision techniques have provided technical solutions to promote production efficiency and reduce labor costs in aquaculture and marine resource surveys. Traditional manual surveys are being replaced by advanced intelligent technologies. However, underwater object detection and recognition are suffering from the image distortion and degradation issues. In this work, automatic monitoring of sea cucumber in natural conditions is implemented based on a state-of-the-art object detector, YOLOv7. To depress the image distortion and degradation issues, image enhancement methods are adopted to improve the accuracy and stability of sea cucumber detection across multiple underwater scenes. Five well-known image enhancement methods are employed to improve the detection performance of sea cucumber by YOLOv7 and YOLOv5. The effectiveness of these image enhancement methods is evaluated by experiments. Non-local image dehazing (NLD) was the most effective in sea cucumber detection from multiple underwater scenes for both YOLOv7 and YOLOv5. The best average precision (AP) of sea cucumber detection was 0.940, achieved by YOLOv7 with NLD. With NLD enhancement, the APs of YOLOv7 and YOLOv5 were increased by 1.1% and 1.6%, respectively. The best AP was 2.8% higher than YOLOv5 without image enhancement. Moreover, the real-time ability of YOLOv7 was examined and its average prediction time was 4.3 ms. Experimental results demonstrated that the proposed method can be applied to marine organism surveying by underwater mobile platforms or automatic analysis of underwater videos.

A Simple and Effective Visual Fluorescent Sensing Paper-Based Chip for the Ultrasensitive Detection of Mercury Ions in Environmental Water.

Traces of mercury ions in environmental water can harm humans and animals. Paper-based visual detection methods have been widely developed for the rapid detection of mercury ions; however, existing methods are not sensitive enough to be used in real environments. Here, we developed a novel, simple and effective visual fluorescent sensing paper-based chip for the ultrasensitive detection of mercury ions in environmental water. CdTe-quantum-dots-modified silica nanospheres were firmly absorbed by and anchored to the fiber interspaces on the paper's surface to effectively avoid the unevenness caused by liquid evaporation. The fluorescence of quantum dots emitted at 525 nm can be selectively and efficiently quenched with mercury ions, and the ultrasensitive visual fluorescence sensing results attained using this principle can be captured using a smartphone camera. This method has a detection limit of 2.83 µg/L and a fast response time (90 s). We successfully achieved the trace spiking detection of seawater (from three regions), lake water, river water and tap water with recoveries in the range of 96.8-105.4% using this method. This method is effective, low-cost, user-friendly and has good prospects for commercial application. Additionally, the work is expected to be utilized in the automated big data collection of large numbers of environmental samples.

Three dimensionally printed nitrocellulose-based microfluidic platform for investigating the effect of oxygen gradient on cells.

In this article, we present a novel nitrocellulose-based microfluidic chip with 3-dimensional (3D) printing technology to study the effect of oxygen gradient on cells. Compared with conventional polydimethylsiloxane (PDMS) chips of oxygen gradient for cell cultures that can only rely on fluorescence microscope analysis, this hybrid nitrocellulose-based microfluidic platform can provide a variety of analysis methods for cells, including flow cytometry, western blot and RT-PCR, because the nitrocellulose-based chips with cells can be taken out from the growth chambers of 3D printed microfluidic chip and then used for cell collection or lysis. These advantages allow researchers to acquire more information and data on the basic biochemical and physiological processes of cell life. The effect of oxygen gradient on the zebrafish cells (ZF4) was used as a model to show the performance and application of our platform. Hypoxia caused the increase of intercellular reactive oxygen species (ROS) and accumulation of hypoxia-inducible factor 1α (HIF-1α). Hypoxia stimulated the transcription of hypoxia-responsive genes vascular endothelial growth factor (VEGF) and induced cell cycle arrest of ZF4 cells. The established platform is able to obtain more information from cells in response to different oxygen concentration, which has potential for analyzing the cells under a variety of pathological conditions.

A Cost-Effective In Situ Zooplankton Monitoring System Based on Novel Illumination Optimization.

A cost-effective and low-power-consumption underwater microscopic imaging system was developed to capture high-resolution zooplankton images in real-time. In this work, dark-field imaging was adopted to reduce backscattering and background noise. To produce an accurate illumination, a novel illumination optimization scheme for the light-emitting diode (LED) array was proposed and applied to design a lighting system for the underwater optical imaging of zooplankton. A multiple objective genetic algorithm was utilized to find the best location of the LED array, which resulted in the specific illumination level and most homogeneous irradiance in the target area. The zooplankton imaging system developed with the optimal configuration of LEDs was tested with Daphnia magna under laboratory conditions. The maximal field of view was 16 mm × 13 mm and the optical resolution was 15 µm. The experimental results showed that the imaging system developed could capture high-resolution and high-definition images of Daphnia. Subsequently, Daphnia individuals were accurately segmented and their geometrical characters were measured by using a classical image processing algorithm. This work provides a cost-effective zooplankton measuring system based on an optimization illumination configuration of an LED array, which has a great potential for minimizing the investment and operating costs associated with long-term in situ monitoring of the physiological state and population conditions of zooplankton.

A graphene oxide/gold nanoparticle-based amplification method for SERS immunoassay of cardiac troponin I.

Cardiac troponin I (cTnI) was considered as the "gold standard" for acute myocardial infarction (AMI) diagnosis owing to its superior cardiac specificity for cardiac damage and showing little or no changes in patients with a skeletal muscle disease or trauma. Herein, a new signal amplification surface-enhanced Raman scattering (SERS) platform was developed for recognition and detection of cTnI by using gold nanoparticles (AuNPs), graphene oxide (GO) and magnetic beads (MB). Here, antibody/Raman reporter labeled AuNP-functionalized GO were employed as both SERS nanotags and signal amplification carriers. Monoclonal antibody modified MB were applied as the capture probe and separation agents. In the presence of cTnI, sandwich type immunocomplexes, "capture probe/target/SERS nanotags", were formed through antibody-antigen-antibody interactions. Due to the strong SERS enhancement ability of the designed GO/AuNP complexes and a high binding chance between cTnI and the GO/AuNP complexes, the proposed SERS-based immunoassay could selectively detect cTnI with a high sensitivity (detection limit of 5 pg mL-1) and a good linearity was obtained in a range of 0.01-1000 ng mL-1. In addition, this method was also successfully applied for detecting cTnI in serum substitute media with a similar linear range. Furthermore, this strategy can be constructed with different kinds of antibodies and Raman reporters, and thus can be easily used for simultaneous detection of multiple biomarkers. Therefore, this proposed SERS-based signal amplification technique shows strong potential for the clinical diagnosis of AMI disease.

Rotational Paper-Based Microfluidic-Chip Device for Multiplexed and Simultaneous Fluorescence Detection of Phenolic Pollutants Based on a Molecular-Imprinting Technique.

In this study, we first present rotational paper-based microfluidic chips (RPADs) combined with a molecular-imprinting (MIP) technique to detect phenolic pollutants. The proposed rotational paper-based microfluidic chips could implement qualitative and quantitative analysis of two different phenolic contaminants, 4-nitrophenol (4-NP) and 2,4,6-trinitrophenol (TNP), simultaneously. Qualitative and quantitative analysis could be implemented simultaneously through fluorescence-intensity changes depending on the structures of quantum dots combined with a molecular-imprinting technique. Moreover, the rotational paper-based microfluidic chips provide a low cost, flexible, and easy way to operate the entire process conveniently. Under the optimal conditions, the proposed sensors showed high sensitivity and selectivity. Our final experimental results illustrated that the detection limits of 4-NP and TNP in the paper-based quantum-dot MIP (PQ-MIP) RPADs ranged from 0.5 to 20.0 mg/L, with detection limits of 0.097 and 0.071 mg/L, respectively. This novel rotational paper-based microfluidic device shows great potential and versatility for multiplexed, portable, and rapid testing of environmental and biological samples in the future.

Controlling Capillary-Driven Fluid Transport in Paper-Based Microfluidic Devices Using a Movable Valve.

This paper describes a novel strategy for fabricating the movable valve on paper-based microfluidic devices to manipulate capillary-driven fluids. The movable valve fabrication is first realized using hollow rivets as the holding center to control the paper channel in different layer movement that results in the channel's connection or disconnection. The relatively simple valve fabrication procedure is robust, versatile, and compatible with microfluidic paper-based analytical devices (µPADs) with differing levels of complexity. It is remarkable that the movable valve can be convenient and free to control fluid without the timing setting, advantages that make it user-friendly for untrained users to carry out the complex multistep operations. For the performance of the movable valve to be verified, several different designs of µPADs were tested and obtained with satisfactory results. In addition, in the proof-of-concept enzyme-linked immunosorbent assay experiments, we demonstrate the use of these valves in µPADs for the successful analysis of samples of carcino-embryonic antigen, showing good sensitivity and reproducibility. We hope this technique will open new avenues for the fabrication of paper-based valves in an easily adoptable and widely available way on µPADs and provide potential point-of-care applications in the future.

Portable paper-based device for quantitative colorimetric assays relying on light reflectance principle.

This paper presents a novel paper-based analytical device based on the colorimetric paper assays through its light reflectance. The device is portable, low cost (<20 dollars), and lightweight (only 176 g) that is available to assess the cost-effectiveness and appropriateness of the original health care or on-site detection information. Based on the light reflectance principle, the signal can be obtained directly, stably and user-friendly in our device. We demonstrated the utility and broad applicability of this technique with measurements of different biological and pollution target samples (BSA, glucose, Fe, and nitrite). Moreover, the real samples of Fe (II) and nitrite in the local tap water were successfully analyzed, and compared with the standard UV absorption method, the quantitative results showed good performance, reproducibility, and reliability. This device could provide quantitative information very conveniently and show great potential to broad fields of resource-limited analysis, medical diagnostics, and on-site environmental detection.

Phycosphere bacterial disturbance of Saccharina japonica caused by white rot disease relates to seawater nutrients.

In mid-November 2021, there were large areas of white rot disease on cultivated Saccharina japonica in Rongcheng City, China, and diseases were undetected on Sargassum horneri and Porphyra yezoensis. The disturbance direction of bacterial community in the phycosphere after disease outbreak and the relationship with seawater nutrients remain unclear. Here, in situ studies of bacterial community in the non-diseased and diseased areas (Shawo and Dongchu islands) and seawater nutrient levels were carried out. 16S rRNA sequencing showed that the bacterial richness of the studied seaweeds increased in the diseased area. Only in S. japonica, Algitalea outcompeted abundant primary bacteria with probiotic relationships to the host of the non-diseased area, and dominated in the diseased area (17.6% of the total abundance). Nitrogen and phosphorus levels in seawater were 57.8% and 19.6% higher in the non-diseased area than those in the diseased area, respectively, and were strongly correlated with the phycosphere bacteria at the family level of S. japonica. There was no difference in potential pathogenicity between the two areas, while positive signal communications decreased, and nitrogen cycle, chemoheterotrophy, and cellulolysis increased in the diseased area compared to the non-diseased area. Overall, white rot disease caused a structural disturbance in phycosphere bacterial community of S. japonica that related to seawater nutrient levels. Enriched degraders and altered bacterial community functions may exacerbate the disease. This evaluation will provide information for white rot disease management to prevent and mitigate the occurrence of S. japonica outbreaks.

Portable instrument based on color sensor chip for on-site rapid detection of dissolved sulfides in environmental water samples.

To ensure real-time validity of the detection of unstable toxic environmental pollutants, such as dissolved sulfides, we developed a portable on-site rapid analysis instrument. Through novel design of the color sensor chip-based core sensing components and the conversion between color signal and absorbance by Lambert's law, the instrument showed great performance for rapid (within 3 min) and sensitive on-site detection of sulfides in the environment. It is easy to achieve user-friendly, sample in-answer out, one-stop operation due to the touch-screen-integrated user interface of the instrument's data terminal. The detection limit of this method is 2.24 µg/L, the linear operation range is 0-1000 µg/L, and the coefficient of determination is 0.999. This instrument has been successfully applied to the on-site determination of sulfides in the Yellow River Delta and the Yantai Guangdang River in China. The portable instrument showed excellent anti-interference, good stability, and simple operation, which showed great prospects for the on-site rapid analysis of unstable targets in the environment.

Fabrication and application of molecularly imprinted polymer doped carbon dots coated silica stationary phase.

Facing the difficulties in chromatographic separation of polar compounds, this investigation devotes to developing novel stationary phase. Molecularly imprinted polymers (MIPs) have aroused wide attention, owing to their outstanding selectivity, high stability, and low cost. In this work, a novel stationary phase based on carbon dots (CDs), MIP layer, and silica beads was synthesized to exploit high selectivity of MIPs, excellent physicochemical property of CDs, and outstanding chromatographic performances of silica microspheres simultaneously. The MIP doped CDs coated silica (MIP-CDs/SiO2) stationary phase was systematically characterized by scanning electron microscopy (SEM), Brunauer-Emmett-Teller (BET) surface area measurement, and carbon elemental analysis. Furthermore, the chromatographic performance of the MIP-CDs/SiO2 column was thoroughly assessed by using a wide variety of compounds (including nucleosides, sulfonamides, benzoic acids, and some other antibiotics). Meanwhile, the separation efficiency of the MIP-CDs/SiO2 stationary phase was superior to other kinds of stationary phases (e.g. nonimprinted NIP-CDs/SiO2, MIP/SiO2, and C18-SiO2). The results demonstrated that MIP-CDs/SiO2 column exhibited best performance in terms of chromatographic separation. For all tested compounds, the resolution value was not less than 1.60, and the column efficiency of MIP-CDs/SiO2 for thymidine was 22,740 plates/m. The results further indicate that the MIP-CDs/SiO2 column can combine the good properties of MIP, CDs, with those of silica microbeads. Therefore, the developed MIP-CDs/SiO2 stationary phase can be applied in the separation science and chromatography-based techniques.

Profiling of the spatiotemporal distribution, risks, and prioritization of pharmaceuticals and personal care products in coastal waters of the northern Yellow Sea, China.

Pharmaceuticals and personal care products (PPCPs) have aroused global concerns due to their ubiquitous occurrence and detrimental effects. The spatiotemporal distributions of 64 PPCPs and their synergetic ecological risks were comprehensively investigated in the seawater of Yantai Bay, and 1 H-benzotriazole (BT), ethenzamide, phenazone, propyphenazone, 4-hydroxybenzophenone and N, N'-diphenylurea were first determined in the seawater of China. Fifty-six PPCPs were detected and their concentrations were 27.5-182 ng/L, with BT contributing around 58.0%. Higher PPCP concentrations were observed in winter and spring, with the concentrations of antioxidants, analgesic/anti-inflammatory drugs and human-used antibiotics significantly higher in winter, while those of aquaculture-used antibiotics and UV filters significantly higher in summer, which was closely related with their usage patterns. Positive correlations were observed for PPCP concentrations between surface and bottom water, except summer, during which time the weak vertical exchange and varied environmental behaviors among different PPCPs resulted in the distinct compositions and concentrations. Terrestrial inputs and mariculture resulted in higher PPCP concentrations in the area located adjacent to the coast and aquaculture bases. The PPCP mixtures posed medium to high risk to crustaceans, and bisphenol A was identified as a high-risk pollutant that needs special attention.

Dissolved oxygen gradient on three dimensionally printed microfluidic platform for studying its effect on fish at three levels: cell, embryo, and larva.

A simple and low-cost dissolved oxygen gradient platform of three dimensionally (3D) printed microfluidic chip was developed for cultivating cells, embryos, and larvae of fish. "Christmas tree" structure channel networks generated a dissolved oxygen gradient out of two fluids fed to the device. Polydimethylsiloxane (PDMS) membrane with high biocompatibility was used as the substrate for cell culture in the 3D-printed microfluidic chip, which made the cell analysis easy. The embryos and larvae of fish could be cultured directly in the chip, and their development can be observed in real time with a microscope. Using zebrafish as a model, we assessed the effect of different dissolved oxygen on its cells, embryos, and larvae. Hypoxia induced production of reactive oxygen species (ROS) in zebrafish cells, embryos, and larvae, eventually leading to cell apoptosis and developmental impairment. Hypoxia also increased nitric oxide content in zebrafish cells, which might be a defensive strategy to overcome the adverse effect of hypoxia in fish cells. This is the first platform that could comprehensively investigate the effects of different dissolved oxygen on fish at the cell, embryo, and larva levels, which has great potential in studying the responses of aquatic organisms under different oxygen concentrations.

Chiral molecular imprinting-based SERS detection strategy for absolute enantiomeric discrimination.

Chiral discrimination is critical in environmental and life sciences. However, an ideal chiral discrimination strategy has not yet been developed because of the inevitable nonspecific binding entity of wrong enantiomers or insufficient intrinsic optical activities of chiral molecules. Here, we propose an "inspector" recognition mechanism (IRM), which is implemented on a chiral imprinted polydopamine (PDA) layer coated on surface-enhanced Raman scattering (SERS) tag layer. The IRM works based on the permeability change of the imprinted PDA after the chiral recognition and scrutiny of the permeability by an inspector molecule. Good enantiomer can specifically recognize and fully fill the chiral imprinted cavities, whereas the wrong cannot. Then a linear shape aminothiol molecule, as an inspector of the recognition status is introduced, which can only percolate through the vacant and nonspecifically occupied cavities, inducing the SERS signal to decrease. Accordingly, chirality information exclusively stems from good enantiomer specific binding, while nonspecific recognition of wrong enantiomer is curbed. The IRM benefits from sensitivity and versatility, enabling absolute discrimination of a wide variety of chiral molecules regardless of size, functional groups, polarities, optical activities, Raman scattering, and the number of chiral centers.

A novel polymer-based nitrocellulose platform for implementing a multiplexed microfluidic paper-based enzyme-linked immunosorbent assay.

Nitrocellulose (NC) membranes, as porous paper-like substrates with high protein-binding capabilities, are very popular in the field of point-of-care immunoassays. However, generating robust hydrophobic structures in NC membranes to fabricate microfluidic paper-based analytical devices (µPADs) remains a great challenge. At present, the main method relies on an expensive wax printer. In addition, NC membranes very easy to adhere during the printing process due to electrostatic adsorption. Herein, we developed a facile, fast and low-cost strategy to fabricate µPADs in NC membranes by screen-printing polyurethane acrylate (PUA) as a barrier material for defining flow channels and reaction zones. Moreover, hydrophobic barriers based on UV-curable PUA can resist various surfactant solutions and organic solvents that are generally used in immunoassays and biochemical reactions. To validate the feasibility of this PUA-based NC membrane for immunoassays in point-of-care testing (POCT), we further designed and assembled a rotational paper-based analytical device for implementing a multiplexed enzyme-linked immunosorbent assay (ELISA) in a simple manner. Using the proposed device under the optimal conditions, alpha fetoprotein (AFP) and carcinoembryonic antigen (CEA) could be identified, with limits of detection of 136 pg/mL and 174 pg/mL, respectively, which are below the threshold values of these two cancer biomarkers for clinical diagnosis. We believe that this reliable device provides a promising platform for the diagnosis of disease based on ELISA or other related bioassays in limited settings or remote regions.

The distinct toxicity effects between commercial and realistic polystyrene microplastics on microbiome and histopathology of gut in zebrafish.

As a ubiquitous emerging pollutant, microplastics (MPs) have attracted widespread attention. At this stage, researchers mainly employed commercial MPs (CMPs) as the model particles to explore the toxic effects of MPs. But whether CMPs can reflect the effects of realistic MPs (RMPs) still remains unknown. Herein, the effects of commercial and realistic polystyrene MPs on gut microbiota of zebrafish were compared. Considering MPs co-exist with antibiotics in real environment, we further distinguished the effects of CMPs and RMPs when they co-existed with enrofloxacin (ENR). The results revealed that while both CMPs and RMPs significantly shifted the gut microbiota, CMPs exhibited stronger toxic effects and more severe damage to gut. Furthermore, ENR exhibited a distinct effect with both CMPs and RMPs on gut microbiota, while the addition of CMPs and RMPs significantly alleviated the toxicity of ENR. In addition, analysis via Kyoto Encyclopedia of Genes and Genomes pathway database revealed that seven major level 1 pathways associated with metabolism, information processing and diseases in the microbial community were affected. Taken together, this work is the first to report that CMPs could not represent RMPs in terms of toxicity and other behaviors, reminding people the limits of using CMPs in ecotoxicology studies.

Surface-enhanced Raman scattering labeled nanoplastic models for reliable bio-nano interaction investigations.

Nanoplastics (NPs) have attracted great attention as an emerging pollution. To date, their interaction with biological systems has been studied mostly by using fluorescent-labeled NPs, which suffered from serious drawbacks such as biological autofluorescence interference and false-positive results. Reliable optically labeled NP models are eagerly desired until now. Herein, a novel near-infrared (NIR) surface-enhanced Raman scattering (SERS) labeled NP model was proposed, which gained single-particle ultra-sensitivity, deep tissue detection, multiplex labeling ability, and anti-interference property. More importantly, the NP demonstrated satisfactory in vivo signal stability which completely prevented the positive-false problems. The advantages of the NPs enabled direct, dynamic in vivo behavior imaging study in living zebrafish embryo, adult zebrafish and green vegetable Brassica rapa. It was found for the first time that NPs entered blood circulation system of zebrafish larva via dermal uptake route, which only occurred in a short 48 h-window post-hatch. NPs widely distributed in roots, shoots and leaves of Brassica rapa seedlings germinating and growing in the NP-containing hydroponic culture. Different depths of one root showed varied adsorption capabilities towards NPs with fulvic acid, lipid and sodium dodecyl sulfate eco-coronas. This work provided an ideal tool for reliable bio-NP interaction study for a variety of organisms, which could promote the research of NPs.

Label-free SERS detection of Raman-Inactive protein biomarkers by Raman reporter indicator: Toward ultrasensitivity and universality.

It is still challenging to sensitively detect protein biomarkers via surface-enhanced Raman scattering (SERS) technique owing to their low Raman activity. SERS tag-based immunoassay is usually applied; however, it is laborious and needs specific antibodies. Herein, an ultrasensitive and universal "Raman indicator" sensing strategy is proposed for protein biomarkers, with the aid of a glass capillary-based molecularly imprinted SERS sensor. The sensor consists of an inner SERS substrate layer for signal enhancement and an outer mussel-inspired polydopamine imprinted layer as a recognition element. Imprinted cavities have two missions: first, selectively capturing the target protein, and second, the only passageway of Raman indicator to access SERS substrate. Specific protein recognition means filling imprinted cavities and blocking Raman indicator flow. Thus, the quantity of captured protein can be reflected by the signal decrease of ultra-Raman active indicator molecule. The capillary sensor exhibited specific and reproducible detection at the level down to 4.1 × 10-3 µg L-1, for trypsin enzyme in as-received biological samples without sample preparation. The generality of the mechanism is confirmed by using three different protein models. This platform provides a facile, fast and general route for sensitive SERS detection of Raman inactive biomacromolecules, which offers great promising utility for in situ and fast point-of-care practical bioassay.