Poly(ferrocenylsilane)-Based Redox-Active Artificial Organelles for Biomimetic Cascade Reactions.

Artificial organelles serve as functional counterparts to natural organelles, which are primarily employed to artificially replicate, restore, or enhance cellular functions. While most artificial organelles exhibit basic functions, we diverge from this norm by utilizing poly(ferrocenylmethylethylthiocarboxypropylsilane) microcapsules (PFC MCs) to construct multifunctional artificial organelles through water/oil interfacial self-assembly. Within these PFC MCs, enzymatic cascades are induced through active molecular exchange across the membrane to mimic the functions of enzymes in mitochondria. We harness the inherent redox properties of the PFC polymer, which forms the membrane, to facilitate in-situ redox reactions similar to those supported by the inner membrane of natural mitochondria. Subsequent studies have demonstrated the interaction between PFC MCs and living cell including extended lifespans within various cell types. We anticipate that functional PFC MCs have the potential to serve as innovative platforms for organelle mimics capable of executing specific cellular functions.

Macromolecular Nano-Assemblies for Enhancing the Effect of Oxygen-Dependent Photodynamic Therapy Against Hypoxic Tumors.

In oxygen (O2)-dependent photodynamic therapy (PDT), photosensitizers absorb light energy, which is then transferred to ambient O2, and subsequently cytotoxic singlet oxygen (1O2) is generated. Therefore, the availability of O2 and the utilization efficiency of generated 1O2 are two significant factors that influence the effectiveness of PDT. However, tumor microenvironments (TMEs) characterized by hypoxia and limited utilization efficiency of 1O2 resulting from its short half-life and short diffusion distance significantly restrict the applicability of PDT for hypoxic tumors. To address these challenges, numerous macromolecular nano-assemblies (MNAs) have been designed to relieve hypoxia, utilize hypoxia or enhance the utilization efficiency of 1O2. Herein, we provide a comprehensive review on recent advancements achieved with MNAs in enhancing the effectiveness of O2-dependent PDT against hypoxic tumors.

Polysaccharide/Lipid Nanoconjugates as Alternative Building Blocks for Highly Biocompatible Microcapsules.

Saccharide/lipid nanoconjugates are attractive building blocks for the construction of micro- and nanosized structures because of the roles of glycolipids in human body, courtesy of their intrinsic and functional properties. Herein, nanoconjugates based on dextran and oleic acid (Dex-OA) were synthesized via facile amide-linkage chemistry. The resultant Dex-OA micelles could self-assemble into spherical water-filled microcapsules via a water-in-oil emulsification process. By cross-linking, the microcapsules could be transferred to aqueous media, forming a stable microcapsule dispersion. According to optical and fluorescence microscopy, the microcapsules displayed a spherical morphology, and their synthesis is dependent on the concentration of Dex-OA nanoconjugates. Furthermore, the microcapsules could easily encapsulate and retain fluorescently labeled dextran. This strategy offers a robust and efficient method for the construction of microcapsules from fully natural amphiphilic building blocks with the potential for application in diverse fields such as biomedicine, protocell research, and microreactors.

Hierarchical Structures in Macromolecule-Assembled Synthetic Cells.

Various models of synthetic cells have been developed as researchers have sought to explore the origins of life. Based on the fact that structural complexity is the foundation of higher-order functions, this review focuses on hierarchical structures in synthetic cell models that are inspired by living systems, in which macromolecules are the dominant participants. The underlying advantages and functions provided by biomimetic higher-order structures are discussed from four perspectives, including hierarchical structures in membranes, in the composite construction of membrane-coated artificial cytoplasm, in organelle-like subcellular compartments, as well as in synthetic cell-cell assembled synthetic tissues. In parallel, various feasible driving forces and approaches for the fabrication of such higher-order structures are showcased. Furthermore, both the implemented and potential applications of biomimetic systems, bottom-up biosynthesis, biomedical tissue engineering, and disease therapy are highlighted. This thriving field is gradually narrowing the gap between fundamental research and applied science.

Lipid-coated albumin-paclitaxel nanoparticles loaded with sorcin-siRNA reverse cancer chemoresistance via restoring intracellular calcium ion homeostasis.

Chemoresistance is often a cause of the failure of chemotherapy in cancer treatment. Sorcin (SRI) is a soluble resistance-related calcium-binding protein involved in chemoresistant processes and is overexpressed in many chemoresistant cancer cells, including paclitaxel (PTX)-resistant ovarian cancer. Increased SRI can reduce the concentration of calcium ions in the cytosol and mitochondria and the decrease of calcium ion concentration prevents the occurrence of apoptosis. Here we examined the SRI expression in multiple cancers using a human TissueArray and found that SRI expression was significantly higher in malignant tumor tissues. Furthermore, SRI was overexpressed, while intracellular calcium concentration was decreased, in chemoresistant cancer cells. To restore intracellular calcium homeostasis and overcome chemoresistance, we developed lipid-coated albumin-PTX nanoparticles loaded with SRI-siRNA (LANP-PTX-siSRI) for PTX and SRI-siRNA co-delivery. LANP-PTX-siSRI had dual-target roles in the regulation of SRI and the delivery of PTX into chemoresistant cells. The LANP-PTX-siSRI inhibited the expression of SRI and enhanced intracellular calcium, leading to the induction of apoptosis and the inhibition of the growth of PTX-resistant cancer cells in vitro and in vivo. In addition, the mechanism study revealed that the overexpression of SRI was associated with an impaired TGF-ß signaling pathway. The administration of TGF-ß1 inhibited two calcium-binding proteins SRI and S100A14. In conclusion, our data unveil that restoring intracellular calcium ion homeostasis via reducing SRI expression can reverse chemoresistance. Thus, the fabricated LANP-PTX-siSRI has a potentially therapeutical application.

In Situ Preparation of Composite Redox-Active Micelles Bearing Pd Nanoparticles for the Reduction of 4-Nitrophenol.

Owing to the redox activity of the poly(ferrocenylsilane)-based polymer, several noble metal nanoparticles can be successfully prepared. As reported herein, the in situ preparation of Pd nanoparticles was performed using a redox-active platform of poly(ferrocenylmethylethylthiocarboxylpropylsilane) (PFC) micelles. PFC/Pd nanocomposites (NCs) with Pd nanoparticles uniformly dispersed at the surface of PFC nanospheres were obtained. The morphology of PFC/Pd NCs was further confirmed via high-resolution transmission electron microscopy and X-ray photoelectron spectroscopy. Taking advantage of Pd nanoparticles, the PFC/Pd NCs showed significant catalytic activity during the reduction process of 4-nitrophenol by sodium borohydride. Although PFC micelles themselves showed no catalytic activity, they promoted the catalytic behavior of Pd nanoparticles obviously by anchoring the Pd nanoparticles at their surface to avoid the aggregation and leaching of Pd nanoparticles. In all, PFC/Pd NCs exhibited great potential as a composite nanocatalyst. Moreover, the PFC micelle was found to be a desired platform for nanocatalysts.

Mixed Solvent Method for Improving the Size Uniformity and Cargo-Loading Efficiency of ZIF-8 Nanoparticles.

Zeolitic imidazolate framework-8 (ZIF-8) nanoparticles with tunable diameters and a uniform morphology were constructed in dimethyl sulfoxide (DMSO)/H2O mixed solvents and were further decorated with dextran to improve their stability and biocompatibility. A series of reaction conditions, including the DMSO content in mixed solvents, molar ratio between precursors, growth time, and decoration of dextran, were systematically investigated. Most importantly, it was the union of DMSO and water that achieved the combined merits of both solvothermal and hydrothermal methods, namely, high uniformity and high efficiency, respectively. In addition, numerous properties of these ZIF-8 nanoparticles were subsequently studied, such as the crystal structure, surface properties, and porosity. Furthermore, composite ZIF-8 nanoparticles encapsulating various functional molecules were also successfully prepared in the same DMSO/H2O mixed solvents, thus laying the foundation for their application as nanocarriers in the biomedical field.

Biofabrication of Gold Nanotriangles Using Liposomes as a Dual Functional Reductant and Stabilizer.

Negatively charged liposomes accomplished both functions as a reducing and stabilizing agent in the synthesis of gold nanotriangles (GNTs). Liposomes are based on a mixture of phospholipids phosphatidylcholine/phosphoglycerol, and they were used as a template phase to perform the GNTs. The method was evaluated under different conditions such as temperature, reaction time, phosphoglycerol chain length, and precursor concentration. Isotropic and anisotropic gold nanoparticles are formed simultaneously during the synthesis. Therefore, by combining centrifugation and depletion flocculation strategies, the sample was concentrated in terms of GNTs from 15% crude to 80% by using sodium dodecyl sulfate (SDS). As a result, a green colored dispersion was obtained containing highly purified, well-defined, negatively charged GNTs, where the edge length of most particles is centered in the range of 60-80 nm with an average thickness of 7.8 ± 0.1 nm. By this purification process, it was possible to highly increase the yield in terms of GNTs. Other surfactants [cetyltrimethylammonium chloride (CTAC), hexadecyltrimethylammonium bromide (CTAB), Tween 20, and dodecyldimethylammonium bromide] were evaluated during the purification stage, and both CTAB and CTAC show similar results to those obtained by using SDS. These GNTs are potential candidates for future applications in molecular imaging, photothermal therapy, drug delivery, biosensing, and photodynamic therapy.

High-Order Assembly toward Polysaccharide-Based Complex Coacervate Nanodroplets Capable of Targeting Cancer Cells.

High-order assembly plays a significant role in the formation of living organisms containing a large number of biomacromolecules and, thus, enlightens the construction of nanomaterials that can load macromolecular payloads at a high efficiency. Herein, by choosing anionic hyaluronic acid (HA) as a model payload, we demonstrated how the electrostatic-interaction-induced high-order assembly can be used to load efficiently biomacromolecules into complex coacervate nanodroplets. The resultant assemblies were primarily composed of HA and cationic chitosan oligosaccharide/dextran (COS/Dex) nanogels and had a controllable structure while also exhibiting biological functionality. HA in the assemblies is capable of targeting CD44-overexpressed tumor cells through CD44-mediated endocytic pathways, which are elucidated herein. Therefore, this study provides a reliable approach for the efficient loading of macromolecular payloads into complex coacervate nanodroplets via electrostatic-attraction-induced high-order assembly.

Novel Hybrid Dextran-Gadolinium Nanoparticles as High-relaxivity T1 Magnetic Resonance Imaging Contrast Agent for Mapping the Sentinel Lymph Node.

OBJECTIVES: To assess the applicability of a novel hybrid dextran-gadolinium nanoparticles (NPs) as high-relaxivity T1 magnetic resonance imaging (MRI) contrast agent for mapping the sentinel lymph node (SLN). METHODS: Dextran-bis-acrylamide-polyacrylic acid (Dex-MBA-PAA) NPs were synthesized through a self-assembly assisted approach and complexed with multiple chelated gadolinium (Gd) (III) ions. After their characterization was validated, they were used to mapping SLNs by MRI in Wistar rats, and their biosafety was evaluated. RESULTS: Dextran-MBA-polyacrylic acid-Gd NPs have suitable particle size and much higher longitudinal relaxivity (r1) than that of commonly used clinical MRI contrast agents (eg, gadopentetic acid dimeglumine salt injection). The in vivo T1-weighted MRI results revealed their effectiveness at mapping SLNs. And their biological safety was also verified. CONCLUSIONS: Dextran-MBA-polyacrylic acid-Gd NPs were synthesized and validated by in vitro and in vivo experiments for their ability to visualize SLNs by MRI with accurate positioning and excellent biosafety, and they have great potential for clinical SLN mapping.

Self-assembled Fe3O4/polymer hybrid microbubble with MRI/ultrasound dual-imaging enhancement.

An Fe3O4 nanoparticle/polymer hybrid microbubble was developed using a facile self-assembly approach. This approach involves two steps, including the initial fabrication of the iron oxide nanoparticle (IONP)/polymer hybrid microcapsules via self-assembly and a subsequent gas-filling process to yield the final microbubbles. Both in vitro and in vivo experiments demonstrated that the composite gas-filled microbubbles exhibit excellent T2-weighted magnetic resonance imaging (MRI) enhancement as well as ultrasound (US) imaging enhancement capabilities. Besides, this flexible approach allows the facile control of the microbubbles' size and thus the imaging capabilities of the microbubbles through the tuning of the molar ratio between the precursors.

Acid-Responsive Decomposable Nanomedicine Based on Zeolitic Imidazolate Frameworks for Near-Infrared Fluorescence Imaging/Chemotherapy Combined Tumor Theranostics.

Zeolitic imidazolate framework-8 (ZIF-8) nanoparticles (NPs) are gaining traction in tumor theranostics for their effectiveness in encapsulating both imaging agents and therapeutic drugs. While typically, similar hydrophilic molecules are encapsulated in either pure aqueous or organic environments, few studies have explored co-encapsulation of chemotherapeutic drugs and imaging agents with varying hydrophilicity and, consequently, constructed multifunctional ZIF-8 composite NPs for acid-responsive, near-infrared fluorescence imaging/chemotherapy combined tumor theranostics. Here, we present a one-pot method for the synthesis of uniform Cy5.5&DOX@ZIF-8 nanoparticles in mixed solvents, efficiently achieving simultaneous encapsulation of hydrophilic doxorubicin (DOX) and hydrophobic Cyanine-5.5 (Cy5.5). Surface decoration with dextran (Dex) enhanced colloidal stability and biocompatibility. The method significantly facilitated co-loading of Cy5.5 dyes and DOX drugs, endowing the composite NPs with notable fluorescent imaging capabilities and pH-responsive chemotherapy capacities. In vivo near-infrared fluorescence (NIRF) imaging in A549 tumor-bearing mice demonstrated significant accumulation of Cy5.5 at tumor sites due to enhanced permeability and retention (EPR) effects, with fluorescence intensities approximately 48-fold higher than free Cy5.5. Enhanced therapeutic efficiency was observed in composite NPs compared to free DOX, validating tumor-targeted capability. These findings suggest ZIF-8-based nanomedicines as promising platforms for multifunctional tumor theranostics.

Self-assembly of stabilized droplets from liquid-liquid phase separation for higher-order structures and functions.

Dynamic microscale droplets produced by liquid-liquid phase separation (LLPS) have emerged as appealing biomaterials due to their remarkable features. However, the instability of droplets limits the construction of population-level structures with collective behaviors. Here we first provide a brief background of droplets in the context of materials properties. Subsequently, we discuss current strategies for stabilizing droplets including physical separation and chemical modulation. We also discuss the recent development of LLPS droplets for various applications such as synthetic cells and biomedical materials. Finally, we give insights on how stabilized droplets can self-assemble into higher-order structures displaying coordinated functions to fully exploit their potentials in bottom-up synthetic biology and biomedical applications.

DNA as a universal chemical substrate for computing and data storage.

DNA computing and DNA data storage are emerging fields that are unlocking new possibilities in information technology and diagnostics. These approaches use DNA molecules as a computing substrate or a storage medium, offering nanoscale compactness and operation in unconventional media (including aqueous solutions, water-in-oil microemulsions and self-assembled membranized compartments) for applications beyond traditional silicon-based computing systems. To build a functional DNA computer that can process and store molecular information necessitates the continued development of strategies for computing and data storage, as well as bridging the gap between these fields. In this Review, we explore how DNA can be leveraged in the context of DNA computing with a focus on neural networks and compartmentalized DNA circuits. We also discuss emerging approaches to the storage of data in DNA and associated topics such as the writing, reading, retrieval and post-synthesis editing of DNA-encoded data. Finally, we provide insights into how DNA computing can be integrated with DNA data storage and explore the use of DNA for near-memory computing for future information technology and health analysis applications.

NIR-emitting quantum dot-encoded microbeads through membrane emulsification for multiplexed immunoassays.

NIR-emitting CdSeTe/CdS/ZnS core/shell/shell QD-encoded microbeads are combined with common flow cytometry with one laser for multiplexed detection of hepatitis B virus (HBV). A facile one-pot synthetic route is developed to prepare CdSeTe/CdS/ZnS core/shell/shell QDs with high photoluminescence quantum yield and excellent stability in liquid paraffin, and a Shirasu porous glass (SPG) membrane emulsification technique is applied to incorporate the QDs into polystyrene-maleic anhydride (PSMA) microbeads to obtain highly fluorescent QD-encoded microbeads. The relatively wide NIR photoluminescence full width half maximum of the CdSeTe/CdS/ZnS QDs is used to develop a 'single wavelength' encoding method to obtain different optical codes by changing the wavelengh and emission intensity of the QDs incorporated into the microbeads. Moreover, a detection platform combining NIR-emitting CdSeTe/CdS/ZnS QD-encoded microbeads and Beckman Coulter FC 500 flow cytometry with one laser of 488 nm is successfully used to conduct a 2-plex hybridization assay for hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg), and a 3-plex hybridization assay for hepatitis B surface antibody (HBsAb), hepatitis B e antibody (HBeAb), and hepatitis B core antibody (HBcAb), which suggests the promising application of NIR QD-encoded microbeads for multiplex immunoassays.

Signal Transduction in Artificial Cells.

In recent years, significant progress has been made in the emerging field of constructing biomimetic soft compartments with life-like behaviors. Given that biological activities occur under a flux of energy and matter exchange, the implementation of rudimentary signaling pathways in artificial cells (protocells) is a prerequisite for the development of adaptive sense-response phenotypes in cytomimetic models. Herein, recent approaches to the integration of signal transduction modules in model protocells prepared by bottom-up construction are discussed. The approaches are classified into two categories involving invasive biochemical signals or non-invasive physical stimuli. In the former mechanism, transducers with intrinsic recognition capability respond with high specificity, while in the latter, artificial cells respond through intra-protocellular energy transduction. Although major challenges remain in the pursuit of a sophisticated artificial signaling network for the orchestration of higher-order cytomimetic models, significant advances have been made in establishing rudimentary protocell communication networks, providing novel organizational models for the development of life-like microsystems and new avenues in protoliving technologies.

Single-Cell Diagnosis of Cancer Drug Resistance through the Differential Endocytosis of Nanoparticles between Drug-Resistant and Drug-Sensitive Cancer Cells.

Single-cell diagnosis of cancer drug resistance is highly relevant for cancer treatment, as it can be used to identify the subpopulations of drug-resistant cancer cells, reveal the sensitivity of cancer cells to treatment, and monitor the progress of cancer drug resistance. However, simple and effective methods for cancer drug resistance detection at the single-cell level are still lacking in laboratory and clinical studies. Inspired by the fact that nanoparticles with diverse physicochemical properties would generate distinct and specific interactions with drug-resistant and drug-sensitive cancer cells, which have distinctive molecular signatures, here, we have synthesized a library of fluorescent nanoparticles with various sizes, surface charges, and compositions (SiO2 nanoparticles (SNPs), organic PS-co-PAA nanoparticles (ONPs), and ZIF-8 nanoparticles (ZNPs)), thus demonstrating that the composition has a critical influence on the interaction of nanoparticles with drug-resistant cancer cells. Furthermore, the clathrin/caveolae-independent endocytosis of ZNPs together with the P-glycoprotein-related decreased cell membrane fluidity resulted in a lower cellular accumulation of ZNPs in drug-resistant cancer cells, consequently causing the distinct cellular accumulation of ZNPs between the drug-resistant and drug-sensitive cancer cells. This difference was further quantified by detecting the fluorescence signals generated by the accumulation of nanoparticles at the single-cell level via flow cytometry. Our findings provide another insight into the nanoparticle-cell interactions and offer a promising platform for the diagnosis of cancer drug resistance of various cancer cells and clinical cancer samples at the single-cell level.

Efficient incorporation of quantum dots into porous microspheres through a solvent-evaporation approach.

Quantum dot (QD)-encoded microspheres play an important role in suspension arrays by acting as supports for various reactions between biomolecules. With regard to QD-encoded microspheres utilized in suspension arrays, three key requirements are controllable size, abundant surface functional groups, and especially excellent fluorescence properties. In this paper, narrowly dispersed poly(styrene-co-divinylbenzene-co-methylacrylic acid) (PSDM) microspheres with specific size, surface carboxyl groups, and porous structures were synthesized by seeded copolymerization. In order to improve the incorporation efficiency of QDs within microspheres, we developed a swelling-evaporation approach in which the swelling process was combined with gradual evaporation of the solvent and thus gradual concentration of QDs in the dispersion solution. This approach was demonstrated to be an efficient method for improving the fluorescence intensity of resultant microspheres compared with the use of swelling alone. Moreover, the porous structure was shown to aid the penetration of QDs into the interiors of the microspheres. Through this approach, microspheres encoded with either single or multiple wavelength-emitting QDs were fabricated effectively. The suspension immunoassays were then founded based on the QD-encoded microspheres, by coating mouse antihuman chorionic gonadotropin as the probe for goat antimouse IgG detection. The positive results determined by Luminex 100 and the low cytotoxicity of the QD-encoded microspheres demonstrated their great potential in suspension arrays.

Monodisperse ZIF-8@dextran nanoparticles co-loaded with hydrophilic and hydrophobic functional cargos for combined near-infrared fluorescence imaging and photothermal therapy.

Impressive developments have been achieved with the use of zeolitic imidazolate framework-8 (ZIF-8) as nanocarriers for tumor theranostics in recent decades by incorporating imaging agents and therapeutic drugs within ZIF-8. However, the simultaneous immobilization of hydrophilic and hydrophobic functional molecules into ZIF-8 nanoparticles in water or organic solvents still presents a daunting challenge. Herein, we developed a new synthesis/encapsulation two-in-one (denoted as one-pot) approach to synthesize uniform dextran-modified Cy5.5&ICG@ZIF-8-Dex nanoparticles in DMSO/H2O solvent mixtures, which enabled the simultaneous encapsulation of hydrophilic indocyanine green (ICG) and hydrophobic cyanine-5.5 (Cy5.5) during the same step. It was confirmed that the one-pot approach in this mixed solvents facilitated the loading of ICG and Cy5.5 molecules. Moreover, the encapsulation of Cy5.5 and ICG within ZIF-8 nanoparticles endowed them with fluorescence imaging capability and photothermal conversion capacity, respectively. The in vivo near-infrared (NIR) fluorescent images of A549-bearing mice injected with Cy5.5&ICG@ZIF-8-Dex demonstrated sufficient accumulations of Cy5.5 at tumor sites due to the enhanced permeability and retention effect. Most impressively, the fluorescent intensity of Cy5.5&ICG@ZIF-8-Dex at tumor site was approximately 40-fold higher than that of free Cy5.5. Additionally, the results of in vivo infrared imaging and photothermal therapy of Cy5.5&ICG@ZIF-8-Dex showed enhanced therapeutic efficiency in comparison with free ICG, further confirming its tumor-targeting capability and photothermal capacity. Therefore, this multifunctional system based on ZIF-8 nanocarriers offered a potential nanoplatform for tumor-targeting theranostics, thus broadening the synthesis and applications of ZIF-8 composite nanoparticles for NIR fluorescence imaging and photothermal therapy in the biomedical field. STATEMENT OF SIGNIFICANCE: Simultaneous immobilization of hydrophilic and hydrophobic molecules into ZIF-8 nanoparticles still remains a daunting challenge. Therefore, we have developed a new synthesis/encapsulation two-in-one approach to synthesize uniform Cy5.5&ICG@ZIF-8-Dex composite nanoparticles in DMSO/H2O solvent mixtures, which enabled the simultaneous encapsulation of hydrophilic indocyanine green (ICG) and hydrophobic cyanine-5.5 (Cy5.5) functional molecules during a single step. The results showed that the co-loading of Cy5.5 and ICG within the ZIF-8 nanoparticles endowed them with a remarkable fluorescence imaging capability and photothermal conversion capacity. Based on their enhanced convenience and efficacy to simultaneously encapsulate hydrophilic and hydrophobic molecules, the multifunctional nanocarriers that were prepared in the DMSO/H2O mixed solvents provide a potential nanoplatform toward fluorescence imaging and photothermal therapy for tumor theranostics.

Multidimensional Quantitative Measurement of Cancer Chemoresistance through Differential ZIF-8 Nanoparticle Cellular Retention.

Chemoresistance of cancer cells is conventionally quantified by half-maximal inhibitory concentration (IC50) or multidrug resistance gene 1 (MDR1) values, but these metrics can only reflect the overall drug resistance level of a cancer cell line. Meanwhile, the multidimensional evaluation of both the heterogeneity in a cell line and the drug resistance degree of each cell still presents a daunting challenge. We report here that the cellular heterogeneity, cellular cross contamination, and the proportion of chemoresistant cancer cells can be visualized via flow cytometry through the differential cellular retention of fluorescent ZIF-8 nanoparticles. In addition, we show that the degree of drug resistance exhibited by each cell subpopulation can be quantified by differing fluorescence of the drug-resistant and drug-sensitive cells in the corresponding flow cytometry profile, and the quantified metric S is highly consistent with the MDR1 expression results. Importantly, this novel strategy is applicable to various cancer cell lines, thus demonstrating a universal diagnosis platform for multidimensional, quantitative, and highly efficient diagnosis of cancer chemoresistance.