Jingyu Shi

A fluorescence resonance energy transfer (FRET) biosensor based on graphene quantum dots (GQDs) and gold nanoparticles (AuNPs) for the detection of mecA gene sequence of Staphylococcus aureus

In this work, a novel fluorescence resonance energy transfer (FRET) biosensor based on graphene quantum dots (GQDs) and gold nanoparticles (AuNPs) pairs was developed for Staphylococcus aureus specific gene sequence detection. This FRET biosensor platform was realized by immobilization of capture probes on GQDs and conjugation of reporter probes on AuNPs. Target oligos then co-hybridized with capture probes and reporter probes to form a sandwich structure which brought GQDs and AuNPs to close proximity to trigger FRET effect. The fluorescence signals before and after addition of targets were measured and the fluorescence quenching efficiency could reach around 87% with 100 nM target oligo. The limit of detection (LOD) of this FRET biosensor was around 1 nM for S.aureus gene detection. Experiments with both single-base mismatched oligos and double-base mismatched oligos demonstrated the good sequence selectivity of this FRET biosensor.

Nanoparticle based fluorescence resonance energy transfer (FRET) for biosensing applications

In the past few decades, Förster resonance energy transfer (FRET) has been used as a powerful tool for providing nanoscale information in many biosensing and bioanalysis applications. The performance of FRET assays is mainly dependent on the design of donor and acceptor pairs. Recently, a series of nanoparticles start to be used in FRET assays including semiconductor quantum dots (QDs), graphene quantum dots (GQDs), upconversion nanoparticles (UCNPs), gold nanoparticles (AuNPs) and graphene oxide (GO). The rapid pace of development in nanoparticles provides a lot of opportunities to revolutionize FRET techniques. Many nanoparticle based FRET assays have also been developed for various biosensing applications with higher sensitivity and better stability compared with traditional organic fluorophore based FRET assays. This article reviews the recent progress of nanoparticle FRET assays and their applications in biosensing area.

A graphene oxide based fluorescence resonance energy transfer (FRET) biosensor for ultrasensitive detection of botulinum neurotoxin A (BoNT/A) enzymatic activity

Botulinum neurotoxins (BoNTs) are among the most potent toxic bacterial proteins for humans, which make them potential agents for bioterrorism. Therefore, an ultrasensitive detection of BoNTs and their active states is in great need as field-deployable systems for anti-terrorism applications. We report the construction of a novel graphene oxide (GO)-peptide based fluorescence resonance energy transfer (FRET) biosensor for ultrasensitive detection of the BoNT serotype A light chain (BoNT-LcA) protease activity. A green fluorescence protein (GFP) modified SNAP-25 peptide substrate (SNAP-25-GFP) was optimally designed and synthesized with the centralized recognition/cleavage sites. This FRET platform was constructed by covalent immobilization of peptide substrate on GO with BSA passivation which have advantages of low non-specific adsorption and high stability in protein abundant solution. BoNT-LcA can specifically cleave SNAP-25-GFP substrate covalently immobilized on GO to release the fragment with GFP. Based on fluorescence signal recovery measurement, the target BoNT-LcA was detected sensitively and selectively with the linear detection range from 1fg/mL to 1pg/mL. The limit of detection (LOD) for BoNT-LcA is around 1fg/mL.

Graphene and graphene-like two-denominational materials based fluorescence resonance energy transfer (FRET) assays for biological applications.

In the past decades, Förster resonance energy transfer (FRET) has been applied in many biological applications to reveal the biological information at the nanoscale. Recently, graphene and graphene-like two-dimensional (2D) nanomaterials started to be used in FRET assays as donors or acceptors including graphene oxide (GO), graphene quantum dot (GQD), graphitic-carbon nitride nanosheets (g-C3N4) and transition metal dichalcogenides (e.g. MoS2, MnO2, and WS2). Due to the remarkable properties such as large surface to volume ratio, tunable energy band, photoluminescence and excellent biocompatibility, these 2D nanomaterials based FRET assays have shown great potential in various biological applications. This review summarizes the recent development of graphene and graphene-like 2D nanomaterials based FRET assays in applications of biosensing, bioimaging, and drug delivery monitoring.

A graphene quantum dot@Fe3O4@SiO2 based nanoprobe for drug delivery sensing and dual-modal fluorescence and MRI imaging in cancer cells

A novel graphene quantum dot (GQD)@Fe3O4@SiO2 based nanoprobe was reported for targeted drug delivery, sensing, dual-modal imaging and therapy. Carboxyl-terminated GQD (C-GQD) was firstly conjugated with Fe3O4@SiO2 and then functionalized with cancer targeting molecule folic acid (FA). DOX drug molecules were then loaded on GQD surface of Fe3O4@SiO2@GQD-FA nanoprobe via pi-pi stacking, which resulted in Fe3O4@SiO2@GQD-FA/DOX conjugates based on a FRET mechanism with GQD as donor molecules and DOX as acceptor molecules. Meanwhile, we successfully performed in vitro MRI and fluorescence imaging of living Hela cells and monitored intracellular drug release process using this Fe3O4@SiO2@GQD-FA/DOX nanoprobe. Cell viability study demonstrated the low cytotoxicity of Fe3O4@SiO2@GQD-FA nanocarrier and the enhanced therapeutic efficacy of Fe3O4@SiO2@GQD-FA/DOX nanoprobe for cancer cells. This luminomagnetic nanoprobe will be a potential platform for cancer accurate diagnosis and therapy.

A hydrophilic polymer based microfluidic system with planar patch clamp electrode array for electrophysiological measurement from cells.

This paper presents a microfluidic planar patch clamp system based on a hydrophilic polymer poly(ethylene glycol) diacrylate (PEGDA) for whole cell current recording. The whole chip is fabricated by UV-assisted molding method for both microfluidic channel structure and planar electrode partition. This hydrophilic patch clamp chip has demonstrated a relatively high gigaseal success rate of 44% without surface modification compared with PDMS based patch clamp devices. This chip also shows a capability of rapid intracellular and extracellular solution exchange with high stability of gigaseals. The capillary flow kinetic experiments demonstrate that the flow rates of PEGDA microfluidic channels are around two orders of magnitude greater than those for PDMS-glass channels with the same channel dimensions. This hydrophilic polymer based patch clamp chips have significant advantages over current PDMS elastomer based systems such as no need for surface modification, much higher success rate of cell gigaseals and rapid solution exchange with stable cell gigaseals. Our results indicate the potential of these devices to serve as useful tools for pharmaceutical screening and biosensing tasks.

One-Step in Situ Detection of miRNA-21 Expression in Single Cancer Cells Based on Biofunctionalized MoS2 Nanosheets

Here, we report the one-step in situ detection of targeted miRNAs expression in single living cancer cells via MoS2 nanosheet-based fluorescence on/off probes. The strategy is based on the folic acid (FA)-poly(ethylene glycol)-functionalized MoS2 nanosheets with adsorbed dye-labeled single-stranded DNA (ssDNA). Once the nanoprobes are internalized into cancer cells, the hybridization between the probes and target miRNA results in the detachment of dye-labeled ssDNA from MoS2 nanosheets surface, leading to the green fluorescence recovery. In this nanoprobe, MoS2 nanosheets offer advantages of high fluorescence quenching efficiency and extremely low toxicity. The FA conjugation could protect the probes and improve cancer cell transfection efficiency. The ability of this nanoprobe for endogenous miRNA detection in single living cancer cells is demonstrated for two types of cancer cells with different miRNA-21 expressions (MCF-7 and Hela cells). This functionalized MoS2 nanosheet-based nanoprobes could provide a sensitive and real-time detection of intracellular miRNA detection platform.

Facile preparation of recombinant spider eggcase silk spheres via an HFIP-on-Oil approach

Versatile spider silk proteins have been prepared by various methods in morphology of spheres for functional applications. Inspired from natural spinning process, a facile approach for the fabrication of silk spheres is described. Distinct from the traditional emulsification method, silk spheres were assembled as rapidly as 10 s by using the HFIP-on-Oil method without any surfactants and agitation used. Notably, a series of factors, such as evaporation rate of HFIP, polarity and molecular weight of oils play central roles on the final silk morphologies. With regard to the increase of protein concentrations, the average dimension and size distribution of silk spheres were both increased. Together with present study, silk spheres prepared by other methods were summarized for comparison in drug delivery applications. As a proof-of-concept, silk spheres loaded with Rhodamine B and Doxorubicin were investigated for the potential proteinase-enhanced drug delivery. On the extracellular environment, ethanol-mediated silk spheres exhibited higher resistance against enzymatic degradation of proteinase K when compared with pristine spheres. Under fluorescent detection by the aid of CLSM, proteinase-enhanced release behaviors were further demonstrated through in-vitro experiments within Hela cells. The facile fabrication of spheres with tunable β-sheets establishes a fascinating platform for functional silk-based applications.