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We developed a method of reducing the background fluorescence of samples made from formalin-fixed and paraffin-embedded blocks of the brain of the second-trimester human fetuses. For reducing excess background fluorescence, the samples were subjected to photobleaching using an LED lamp with blue and red emission peaks in the range of visible spectrum in a construction of an original design. The decrease in the background autofluorescence was checked by measuring the intensity of the emitted background fluorescence of the samples and relative abundance of immunopositive structures after immunohistochemical staining. It was found that the proposed method reliably reduced the background fluorescence of the samples, which improved the quality of multicolor immunofluorescence images of the cerebral cortex.

Background Fluorescence imaging using indocyanine green (ICG) enables intraoperatively visualizing liver tumors as fluorescent. This study evaluated the doses and timing of ICG administration for visualizing tumors via fluorescence using near-infrared light camera systems. Methods Consecutive patients who underwent open liver resection for liver tumors from 2016 to 2017 were included. ICG was intravenously injected one-day before surgery at 0.25 mg-intravenous injection (IV), 1.25 mg-IV, 2.5 mg-IV, or 3.75 mg-IV. No additional ICG was administered when patients underwent ICG (0.5 mg/kg) retention test within 10 preoperative days. The ability of fluorescence imaging to enable identifying liver tumors was compared using the PDE-NEO and PINPOINT. Results 154 lesions in 82 patients were assessed. The tumor identification rate of PDE-NEO did not differ significantly among dosages. The positive predictive values of PDE-NEO were significantly lower at 3.75 mg-IV (69.0%) than in the control group (92.0%) (p = 0.036) and at 1.25 mg-IV (88.9%) (p = 0.033). The tumor identification rate of PINPOINT was significantly higher at 3.75 mg-IV (82.4%) than at 1.25 mg-IV (60.0%) (p = 0.035). The positive predictive values of PINPOINT did not significantly differ among dosages. Conclusion Administering 2.5 mg of ICG one-day before surgery can enable identifying tumors via fluorescence imaging when the ICG test was not performed within 10 preoperative days.

Minimising background fluorescence can enhance the visible details of treated fingerprints. Here, a 4-tpt fingerprint powder exhibiting long-lived phosphorescence is applied to this end. The powder was found to suppress background fluorescence, including on challenging surfaces, when using standard forensic equipment and eschewing specialized or bespoke imaging techniques.

The correct development of seeds is a pivotal requirement for species preservation. This process depends on the balance between sensing the environmental stimuli/stressors and hormone-mediated transduction, which results in physiological responses. The red and blue regions of the electromagnetic spectrum are known to influence seed dormancy and germination. Here, we report on the effects induced by the blue (peak at 430 nm) and red (peak at 650 nm) regions of the electromagnetic spectrum on seeds from photo- and skotomorphogenetic capsules developed under white, blue, or red light. Regardless of exposure, seeds from skotomorphogenetic capsules showed an almost absent dormancy in association with altered germination kinetics. Conversely, in seeds from photomorphogenetic capsules, the exposure to the blue region induced skotomorphogenetic-like effects, while the exposure to the whole visible range (350–750 nm), as well as to only the red region, showed a dose-related trend. The observed differences appeared to be dependent on the wavelengths in the red and to be based on transduction mechanisms taking place in fruits. While the molecular bases of this differential effect need to be clarified, the results hint at the role played by different light wavelengths and intensities in seed development and germination. These findings may be relevant for applications in crop production and species safeguarding.

Bio-optical imaging can noninvasively describe specific biochemical reaction events in small animals using endogenous or exogenous imaging reagents to label cells, proteins, or DNA. The fluorescence optical bio-imaging system excites the fluorescent group to a high energy state by excitation light and then generates emission light. However, many substances in the organism will also emit fluorescence after being excited by the excitation light, and the nonspecific fluorescence generated will affect the detection sensitivity. This paper designs and develops a set of high-level biosafety in vivo fluorescence imaging system for small animals suitable for virology research and proposes a target area extraction algorithm for fluorescence images. The fluorescence image target extraction algorithm first maps the nonlinear separation data in the low-dimensional space to the high-dimensional space. Then, based on the analysis of the characteristics of the fluorescent region, a method for discriminating the target fluorescent region based on the two-step entropy function is proposed, and the real target fluorescent region is obtained according to the set connected region. Based on the experiment of collecting and analyzing the in vivo fluorescent images of mice, it is verified that the proposed algorithm can automatically extract the target fluorescent region better than the classical linear model. It shows that the proposed algorithm is less affected by background fluorescence, and the estimated separated spectrum based on this method is closer to the real target spectrum.

The purpose. Study influence of lipo-polysaccharides (LPS) of phytopathogenic bacteria Pseudomonas syringae pv. atrofaciens (McCulloch 1920) Young, Dye and Wilkie 1978 on physiological-biochemical processes in plants of wheat. Methods. Classical microbiologic, biochemical, statistical. Extraction of LPS is realized by 0,85% solution of chloride of sodium. Induction of fluorescence was measured by device «Floratest». Results. Stimulating influence of treatment with LPS of various strains of P. syringae pv. atrofaciens on pigment compound of leaves of plants of summer wheat of grade Pecherianka is determined. Essential lowering background fluorescence in leaves of summer wheat of grade Pecherianka is fixed. At treating with LPS of P. syringae pv. atrofaciens 9417 and P. syringae pv. atrofaciens 9780 — on 38,7 and 37,4% accordingly, at treating with LPS of P. syringae pv. atrofaciens UKM V-1011 — on 16,6%, whereas in alternative with treating with LPS of P. syringae pv. atrofaciens 9400 background fluorescence authentically did not differ from the control. Lipo-polysaccharides of P. syringae pv. atrofaciens 9417 and LPS of P. syringae pv. atrofaciens 9780 oppressed dark phase of photosynthesis. Conclusions. Lipo-polysaccharides of P. syringae pv. atrofaciens cause increase of amount of photosynthetic pigments — chlorophylls a and b at simultaneous lowering in content of carotenoids (for LPS of P. syringae pv. atrofaciens 9417 and LPS of P. syringae pv. atrofaciens 9780) which is accompanied by lowering of the functional activity of photosynthetic apparatus. That spread not only to «light» phase of photosynthesis, but also on efficiency of «dark» responses of cycle of Calvin, reducing their efficiency.

A new long-wavelength fluorescent probe 1 that could specifically identify H2S has been successfully synthesized and applied for imaging H2S in zebrafish. Probe 1 was readily prepared by featuring nitrobenzene as the recognition unit coupled to resorufin. The fluorescence off–on response is based on the fact that H2S can reduce the nitro group to an amino group, followed by the 1,6-rearrangement-elimination and the release of resorufin. By evaluating the application abilities of probe 1 in vivo and vitro, it is shown that probe 1 has high sensitivity and selectivity to H2S, low background fluorescence interference, with a low detection limit of 17.30 μM. Notably, the occurrence of the reaction can be observed by the naked eye, and the color of the solution changes from yellow to pink. More importantly, it is the first time that using paper chips as carrier to detect H2S, which lays a foundation for the practical application of detecting H2S. The excellent analysis and application capabilities of probe 1 make it an effective tool for further application in practice.

This study reports a simple method for visualising and screening latent DNA on tapes using a Diamond™ dye (DD) staining process followed by visualisation using a portable fluorescence microscope. Ten types of tapes were tested, which include those used currently by forensic laboratories for tape-lifting. All ten types were tested for: 1) their auto-fluorescence, 2) properties when stained with DD using three different DD solutions, and 3) PCR inhibition through a direct STR amplification technique. No background fluorescence was noted viewing four types stained with 20 x DD diluted with 0.01% Triton-X. Clear tape (Sellotape®), DNA-free tape (Lovell Surgical Solutions) and brown packing tape (Packmate™) did not inhibit direct STR amplification, while the other six types showed the inhibition of the PCR. The three tapes were selected to assess their cellular material recovery efficiency by comparing the number of stained cells within an entire fingermark before and after tape-lifting. Tape-lifting was performed either once, twice or ten times. The DNA-free tape (Lovell) used in many forensic laboratories gave poor recovery compared to the clear tape (Sellotape®) and brown packing tape (Packmate™). This simple visualising technique allows the cell location to be recorded, and only the area of tape where cells are present to be removed for DNA typing. The process is a simple and effective triage procedure that reduces the processing of tape-lift samples where there are no cells present.

The use of organ-on-a-chip (OOC) devices is a promising alternative to existing cell-based assays and animal testing in drug discovery. A rapid prototyping method with polydimethylsiloxane (PDMS) is widely used for developing OOC devices. However, because PDMS tends to absorb small hydrophobic molecules, the loss of test compounds in cell-based assays and increases in background fluorescence during observation often lead to biased results in cell-based assays. To address this issue, we have fabricated a glass-based OOC device and characterized the medium flow and molecular absorption properties in comparison with PDMS-based devices. Consequently, we revealed that the glass device generated a stable medium flow, restricted the absorption of small hydrophobic molecules, and showed enhanced cell adhesiveness. This glass device is expected to be applicable to precise cell-based assays to evaluate small hydrophobic molecules, for which PDMS devices cannot be applied because of their absorption of small hydrophobic molecules.

It is recognized that cystatin C is an effective marker for monitoring the glomerular filtration rate and the clinical diagnosis of various diseases. In this study, a novel immunochromatographic me...

Unwanted background fluorescence in microscopy can occur when light from fluorescent structures scatters against nearby tissues. Bright cell bodies produce haze that mask nearby dim structures, including neuronal fibers. We have developed a method to eliminate this haze by fitting it to a simple function. This method enables clearer post-imaging visualization of axons in our in vivo imaging of neurons. We are investigating ways to increase the computation speed of our background fitting technique to allow for real-time image improvement. The implementation of our method in real-time imaging will broaden the potential applications of this fitting method.

A wide variety of items are submitted as evidence in a forensic investigation. Identifying the location of DNA on such items is central to maximizing DNA profiling success and thus the ability to link a person of interest to a particular item or crime. Recent publications describe a fluorescent staining method using Diamond™ Dye (DD) to visualize cellular material on the surface of non-porous items (e.g., glass, plastic). However, substrates of varying porosity and background color have not yet been examined. Varying porous substrates (i.e., paper bank note, stamp, cigarette, wooden matchstick, and fabric) and non-porous substrates (i.e., enamel tooth and plastic bank note) were examined for their suitability with the use of DD. To improve the visualization of cellular material on the porous substrates, we also explored two DD diluents and adjusting image contrast. The results suggest the optimal diluent depends on the absorbent nature of the substrate. For example, ethanol was sufficient for visualization on the non-porous substrates, whereas water was better for the porous substrates. While cellular material was detected on the paper bank note, tooth, and stamp, background fluorescence or autofluorescence and surface type of matchstick prevented clear visualization on this substrate. It was also determined that by adjusting the contrast of images for tooth, paper bank note and matchstick aided in the detection of cellular material. Overall, this study extends the use of DD for latent DNA detection to absorbent substrates, highlights the limitations associated with these substrate types, and suggests modifications to improve visualization on these challenging substrates.

Surveillance and detection of polioviruses (PV) remain crucial to monitoring eradication progress. Intratypic differentiation (ITD) using the real-time RT-PCR kit is key to the surveillance workflow, where viruses are screened after cell culture isolation before a subset are verified by sequencing. The ITD kit is a series of real-time RT-PCR assays that screens cytopathic effect (CPE)-positive cell cultures using the standard WHO method for virus isolation. Because ITD screening is a critical procedure in the poliovirus identification workflow, validation of performance of real-time PCR platforms is a core requirement for the detection of poliovirus using the ITD kit. In addition, the continual update and improvement of the ITD assays to simplify interpretation in all platforms is necessary to ensure that all real-time machines are capable of detecting positive real-time signals. Four platforms (ABI7500 real-time systems, Bio-Rad CFX96, Stratagene MX3000P, and the Qiagen Rotor-Gene Q) were validated with the ITD kit and a redesigned poliovirus probe. The poliovirus probe in the real-time RT-PCR pan-poliovirus (PanPV) assay was re-designed with a double-quencher (Zen™) to reduce background fluorescence and potential false negatives. The updated PanPV probe was evaluated with a panel consisting of 184 polioviruses and non-polio enteroviruses. To further validate the updated PanPV probe, the new assay was pilot tested in five Global Polio Laboratory Network (GPLN) laboratories (Madagascar, India, Philippines, Pakistan, and Democratic Republic of Congo). The updated PanPV probe performance was shown to reduce background fluorescence and decrease the number of false positives compared to the standard PanPV probe.

In historical paintings, the detection of low amounts of pigments and dyes by Raman spectroscopy can sometimes be challenging, in particular for fluorescent dyes. This issue can be overcome by using SERS (surface-enhanced Raman spectroscopy) which takes advantage of the properties of nanostructured metal surfaces to quench fluorescence and enhance Raman signals. In this work, silver nanostars (AgNSs) are applied for the first time to real art samples, in particular to painting cross-sections, exploiting their effective SERS properties for pigment identification. The case study is the Madonna della Misericordia of the National Gallery of Parma (Italy). Cross-sections were analyzed at first by optical microscopy, SEM-EDS, and micro-Raman spectroscopy. Unfortunately, in some cross-sections, the application of conventional Raman spectroscopy was hindered by an intense background fluorescence. Therefore, AgNSs were deposited and used as SERS-active agent. The experimentation was successful, allowing us to identify a modern dye, namely copper phthalocyanine. This result, together with the detection of other modern pigments (titanium white) and expert visual examination, allowed to reconstruct the painting history, postdating its realization from the 15th century (according to the Gallery inventory) to 19th century with a heavy role of recent (middle 20th century) restoration interventions.

Hypochlorous acid (HOCl), mainly generated in mitochondria, plays a critical role in various physiological processes. To better understand the role and function of HOCl in mitochondria, herein, we present the design and synthesis of a Mito-QL reporter for probing the HOCl within mitochondria without other interference generated in living cells. Through the combination of TICT/ICT mechanisms, probe Mito-QL, with large stokes shift (203 nm) and low background fluorescence, exhibited excellent sensitivity (900-fold fluorescence enhancement) and selectivity towards HOCl (LOD = 2.4 nM). The co-location experiments confirmed that probe Mito-QL can firstly localize in the mitochondria and then react with HOCl in mitochondria. Also, the probe is capable of imaging endogenous and exogenous HOCl even the generation of HOCl during the ferroptosis of cells, which is beneficial for more efficient application in biological imaging.

A novel rapid method based on a background fluorescence quenching immunochromatographic assay (bFQICA) was established to achieve simultaneously the quantitative detection of 3-methyl-quinoxaline-2-carboxylic acid (MQCA) and quinoxaline-2-carboxylic acid (QCA), which were efficiently extracted and enriched 4 times using immunomagnetic beads from pork. The analysis of field pork samples by bFQICA was in accordance with that of LC-MS/MS; especially, the proposed bFQICA exhibited great advantages in convenience and efficiency, which only takes 30 min for the detection of MQCA and QCA.

Development of an efficient fluorescent probe for sensing hypochlorite in water samples and biological samples is highly demanded. However, the currently reported fluorescent probes for hypochlorite frequently suffered from the problem of high background fluorescence. Herein, based on the combined effect of two different fluorescence quenching groups, we rationally developed a novel fluorescent probe for hypochlorite with extremely low background fluorescence. Notably, due to the doubly quenching groups, the probe could even keep low background fluorescence in a solution with high viscosity. Furthermore, the probe displayed highly sensitive and selective response to hypochlorite, with the detection limits calculated to be 10.5 nM. Practical application demonstrated that the probe was able to quantitatively detect hypochlorite in various water samples with good recovery. Significantly, the probe showed extremely low background fluorescence in living cells and was capable of detecting minor variation of endogenous hypochlorite in RAW 264.7 cells. Moreover, the fluorescence imaging different concentration of hypochlorite in zebrafish has been successfully conducted. The probe developed herein will be widely used as a reliable tool to accurately monitor the variation of hypochlorite in living organism.

150 million people worldwide suffer one or more urinary tract infections (UTIs) annually. UTIs are a significant health burden: societal costs of UTI exceed $3.5 billion in the U.S. alone; 5% of sepsis cases arise from a urinary source; and UTIs are a prominent contributor toward antimicrobial resistance (AMR). Current diagnostic frameworks exacerbate this burden by providing inaccurate and delayed diagnosis. Rapid point-of-care bacterial identification will allow for early precision treatment, fundamentally altering the UTI paradigm. Raman spectroscopy has a proven ability to provide rapid bacterial identification but is limited by weak bacterial signal and a susceptibility to background fluorescence. These limitations may be overcome using surface enhanced Raman spectroscopy (SERS), provided close and consistent application of bacteria to the SERS-active surface can be achieved. Physical filtration provides a means of capturing uropathogens, separating them from the background solution and acting as SERS-active surface. This work demonstrates that filters can provide a means of aggregating bacteria, thereby allowing subsequent enhancement of the acquired Raman signal using metallic nanoparticles. 60 bacterial suspensions of common uropathogens were vacuum filtered onto commercial polyvinylidene fluoride membrane filters and Raman signals were enhanced by the addition of silver nanoparticles directly onto the filter surface. SERS spectra were acquired using a commercial Raman spectrometer (Ocean Optics, Inc.). Principal Component – Linear Discriminant Analysis provided discrimination of infected from control samples (accuracy: 88.75%, 95% CI: 79.22-94.59%, p-value

Tape-lifting is a non-destructive alternative to swabbing for collection of biological materials deposited on surfaces, especially on porous substrates. While there have been a number of studies looking at the efficiency of tapes in terms of recovery and their effect on downstream processes, none has been able to visually monitor cellular material collection. We report on a comparative study of a range of tapes regarding their collection efficiency of cellular material visualised using fluorescence microscopy, their background fluorescence after staining with DD diluted with three different solvent types and inhibition of subsequent PCR using direct PCR. Three of the fourteen tapes were selected for further testing. These were brown packing tape (Packmate™), clear tape (Sellotape®), and DNA-free tape (Lovell Surgical Solutions). These did not inhibit direct STR amplification; the other eleven tapes exhibited either high background fluorescence and/or inhibition of subsequent PCR. The effectiveness of the three tapes for the collection of cellular material was tested after tape-lifting a fingermark once, twice and ten times. The amount of cellular material within fingermarks stained with Diamond™ dye (DD) was recorded using fluorescence microscopy before and after tape-liftings. The DNA-free tape (Lovell) used in many forensic laboratories gave poor recovery compared to the other two tapes. After a tape-lift, an average of 30 % of cellular material was recovered by using DNA-free tape (Lovell), contrasting with an average recovery of 59.5 % for the clear tape (Sellotape®) and 88.8 % for the brown packing tape (Packmate™). The results presented show that standard crime scene sampling tape does collect DNA but is less effective than shop-bought tapes. Full DNA profiles can be generated from all of touched fabric samples that were collected DNA using the three tapes, triaged by DD staining and amplified by direct PCR approach.

The near exponential proliferation of published Raman microspectroscopic applications over the last decade bears witness to the strengths and versatility of this technology. However, laser-induced fluorescence often severely impedes its application to biological samples. Here we report a new approach for near complete elimination of laser-induced background fluorescence in highly pigmented biological specimens (e.g., microalgae) enabling interrogation by Raman microspectroscopy. Our simple chemiphotobleaching method combines mild hydrogen peroxide oxidation with broad spectrum visible light irradiation of the entire specimen. This treatment permits observing intracellular distributions of macromolecular pools, isotopic tracers, and even viral propagation within cells previously not amenable to Raman microspectroscopic examination. Our approach demonstrates the potential for confocal Raman microspectroscopy becoming an indispensable tool to obtain spatially-resolved data on the chemical composition of highly fluorescent biological samples from individual cells to environmental samples.

Recent advances in multiplexed imaging technologies promise to improve the understanding of the functional states of individual cells and the interactions between the cells in tissues. This often requires compilation of results from multiple samples. However, quantitative integration of information between samples is complicated by variations in staining intensity and background fluorescence that obscure biological variations. Failure to remove these unwanted artifacts will complicate downstream analysis and diminish the value of multiplexed imaging for clinical applications. Here, to compensate for unwanted variations, we automatically identify negative control cells for each marker within the same tissue and use their expression levels to infer background signal level. The intensity profile is normalized by the inferred level of the negative control cells to remove between-sample variation. Using a tissue microarray data and a pair of longitudinal biopsy samples, we demonstrated that the proposed approach can remove unwanted variations effectively and shows robust performance., Chang et al. develop an analytical method called RESTORE to control for variations due to technical artifacts in multiplexed imaging. They test their method on a CycIF stained tissue microarray dataset and biopsies processed at different times. Their method can improve the applicability of imaging techniques in diagnostics and inference using unbiased clustering methods.

Genetically-encoded indicators of neuronal activity enable the labeling of a genetically defined population of neurons to optically monitor their activities. However, researchers often find difficulties in identifying relevant signals from excessive background fluorescence. A photoactivatable version of a genetically encoded calcium indicator, sPA-GCaMP6f is a good example of circumventing such an obstacle by limiting the fluorescence to a region of interest defined by the user. Here, we apply this strategy to genetically encoded voltage (GEVI) and pH (GEPI) indicators. Three photoactivatable GEVI candidates were considered. The first one used a circularly-permuted fluorescent protein, the second design involved a Forster resonance energy transfer (FRET) pair, and the third approach employed a pH-sensitive variant of GFP, ecliptic pHluorin. The candidate with a variant of ecliptic pHluorin exhibited photoactivation and a voltage-dependent fluorescence change. This effort also yielded a pH-sensitive photoactivatable GFP that varies its brightness in response to intracellular pH changes.

The work develops a novel method for the detection of bisphenol A (BPA) based on glutathione (GSH)-protected silver sulphide quantum dots (Ag2S QDs), which was simple, fast, non-toxic, sensitive and selective. Ag2S QDs were synthesised with a sulphur–hydrazine hydrate complex as the S2− source and GSH as the stabiliser. Ag2S QDs, near-infrared QDs, offered the advantages over lower background fluorescence and lower signal loss. The fluorescence peak of Ag2S QDs at 650 nm was quenched significantly in the presence of BPA. Good linear correlations were obtained over the concentration range from 0 to 60 μM of BPA under optimised conditions. The developed selective and sensitive method presaged more opportunities for the application in environmental samples.

Circularly polarized luminescence (CPL) is a preferential emission from chiral systems. Introducing CPL to traditional small fluorescence probes will greatly enhance selectivity and sensitivity due to avoiding the background fluorescence. Small-molecule probes are practically useful for bioimaging and sensing but always suffer from weak CPL properties. To optimize chiral perturbation, the study of structure-CPL correlation is an urgent necessity. Here, we investigated the influence of 3, 3′-substituent of BINOL on optical properties and found that iodization could significantly enhance the fluorescence quantum yield (ΦF) and dissymmetry factor (|glum|). Based on this finding, we presented a strategy to design dual-readout probes emiting both robust total fluorescence and CPL upon activation. As an application example, we first designed a pair of thiols probes, R/S-P1, with the fluorescence readout showing good linear response to cysteine, and CPL signal displaying significant enhancement with |glum| from undetectable value to 0.7 × 10−3.

Carbon monoxide (CO) is one of the most important gaseous signal molecules in biological systems. However, the investigation of the functions of CO in living organisms is restricted by the lack of functional molecular tools. To address this critical challenge, we present herein the rational design, synthesis, and in vivo imaging studies of a powerful two-photon excited near-infrared fluorescent probe (1-Ac) for endogenous CO monitoring. The advantageous features of the new probe include high stability, low background fluorescence, large fluorescence enhancement, high sensitivity, and two-photon excitation with emission in the near-infrared region. Significantly, these merits of the probe enable the tracking of endogenous CO in zebrafish embryos and mouse tissues for the first time.

Convenient biosensor for simultaneous multi-analyte detection was increasingly required in biological analysis. A novel flower-like silver (FLS)-enhanced fluorescence/visual bimodal platform for the ultrasensitive detection of multiple miRNAs was successfully constructed for the first time based on the principle of multi-channel microfluidic paper-based analytical devices (µPADs). Fluorophore-functionalized DNA1 (DNA1-N-CDs) was combined with FLS, which was hybridized with quencher-carrying strand (DNA2-CeO2) to form FLS-enhanced fluorescence biosensor. Upon the addition of the target miRNA, the fluorescent intensity of DNA1-N-CDs within the proximity of the FLS was strengthened. The disengaged DNA/CeO2 complex could result in color change after joining H2O2, leading to real-time visual detection of miRNA firstly. If necessary, then the fluorescence method was applied for a accurate determination. In this strategy, the growth of FLS in µPADs not only reduced the background fluorescence but also provided an enrichment of "hot spots" for surface enhanced fluorescence detection of miRNAs. Results also showed versatility of the FLS in the enhancement of sensitivity and selectivity of the miRNA biosensor. Remarkably, this biosensor could detect as low as 0.03fM miRNA210 and 0.06fM miRNA21. Interestingly, the proposed biosensor also possessed good capability of recycling in three cycles upon change of the supplementation of DNA2-CeO2 and visual substitutive device. This method opened new opportunities for further studies of miRNA related bioprocesses and will provide a new instrument for simultaneous detection of multiple low-level biomarkers.

Background Fluorescence live imaging and mathematical modeling have been used in biomedical fields as a novel combined approach in the post-genomic era. The combined approach can potentially reveal novel biological functions that would not be uncovered by conventional molecular gain- and loss-of-function approaches. Highlight We used live imaging-based mathematical modeling and revealed a rhythmic regulatory mode of embryonic tissue development hidden in the stochastic cellular behavior of cell cycle progression. Conclusion The scope of this review is to describe the experimental and theoretical evidence for cell cycle progression as a stochastic molecular and cellular process. In addition, we elucidate the rhythmic mode of tissue development that endows "biological robustness" for reproducible tissue development. Finally, we discuss the potential of the live imaging-based mathematical approach for understanding pathophysiological development in tissues.

Since miRNA21 is a marker of many human tumors, trace detection of miRNA21 contributes to early tumor diagnosis. However, conventional methods are limited by complex sample preprocessing, high costs, or the interference from background fluorescence in biological fluids. Thus, developing simple and cheap miRNA-21 detection methods from biological fluids is very urgent. Our highlights are that: the miRNA-21 detection system based on phosphorescent resonance energy transfer solves the limitations of conventional methods, avoids the interference from the background fluorescence and scattering in biological fluids, eliminates complex sample preprocessing, and is more feasible for miRNA-21 detection in biological fluids. The principle of detection is that: QDs/PDADMAC (QDs+) nanohybrids were prepared from Mn–ZnS RTP QDs and cationic poly(diallyldimethylammonium chloride). Then a miRNA-21 sequence nanoprobe was built via the interaction between the QDs+ and 6-carboxy-X-rhodamine-DNA (ROX-DNA). The PRET between the QDs+ and ROX-DNA could quench the phosphorescence of the QDs+. After the gradual addition of the miRNA21 sequence, owing to the double-stranded structures formed between miRNA21 and ROX-DNA, the ROX-DNA desorbed from the surfaces of the QDs+, which gradually restored the phosphorescence of the QDs+. This nanoprobe had a detection limit of 1.60 nM and a detection range of 8–80 nM. This nanoprobe is more feasible for quantitative detection of miRNA21 in biological samples and has prospects in the medical diagnosis of tumors.

Near-infrared fluorescence imaging (NIRFI) and far-red fluorescence imaging (FRFI) were used to investigate effects of depilation-induced skin pigmentation and diet-induced background fluorescence on fluorescent signal amplitude and lymphatic contraction frequency in C57BL6 mice. Far-red fluorescent signal amplitude, but not frequency, was affected by diet-induced fluorescence, which was removed by feeding the mice an alfalfa-free diet, and skin pigmentation further impacted the amplitude measurement. NIRFI showed minimal background fluorescence; however, skin pigmentation reduced the amplitude of fluorescent signal changes. Therefore, these effects should be taken into account when imaging mice with different states of skin pigmentation and diet-induced background fluorescence in vivo.

Bioluminescent molecular beacons have been developed using a modular design approach that relies on BRET between the bright luciferase NanoLuc and a Cy3 acceptor., Bioluminescent molecular beacons have been developed using a modular design approach that relies on BRET between the bright luciferase NanoLuc and a Cy3 acceptor. While classical molecular beacons are hampered by background fluorescence and scattering, these BRET-beacons allow detection of low pM concentrations of nucleic acids directly in complex media.