Cell Imaging

In the twenty-first century, nanotechnology has become cutting-edge technology. It is interdisciplinary and multidisciplinary, covering numerous fields such as medicine, engineering, biology, physics, material sciences, and chemistry. The present work aims to cover the optical properties, method of preparations, surface modifications, bio-conjugation, characterization, stability, and cytotoxicity of quantum dots (QDs). Articles were reviewed in English literature reporting the pharmaceutical and bio-pharmaceutical aspects of QDs which were indexed in Scopus, web of science, google scholar and PubMed without applying the year of publication criterion. One significant value of utilizing nanotechnology is that one can alter and control the properties in a genuinely unsurprising way to address explicit applications' issues. In science and biomedicine, the usage of functional nanomaterials has been broadly investigated and has become one of the quick-moving and stimulating research d...

In the last decade light sheet fluorescence microscopy techniques, such as selective plane illumination microscopy (SPIM), has become a well established method for developmental biology. However, conventional SPIM architectures hardly permit imaging of certain tissues since the common sample mounting procedure, based on gel embedding, could interfere with the sample morphology. In this work we propose an inverted selective plane microscopy system (iSPIM), based on non-linear excitation, suitable for 3D tissue imaging. First, the iSPIM architecture provides flexibility on the sample mounting, getting rid of the gel-based mounting typical of conventional SPIM, permitting 3D imaging of hippocampal slices from mouse brain. Moreover, all the advantages brought by two photon excitation (2PE) in terms of reduction of scattering effects and contrast improvement are exploited, demonstrating an improved image quality and contrast compared to single photon excitation. The system proposed represents an optimal platform for tissue imaging and it smooths the way to the applicability of light sheet microscopy to a wider range of samples including those that have to be mounted on non-transparent surfaces. Light sheet microscopy has been proven to be an optimal imaging tool in developmental biology, and more generally for the investigation of thick biological samples 1,2. Light sheet microscopy basically relies on the use of two decoupled optical paths for illumination and detection: a widefield based detection path for fast imaging and an orthogonally oriented illumination one, responsible for confining the illumination within a thin planar region in the focal region. Regardless of how the illumination sheet is created, either using a cylindrical lens 1 or rapidly scanning a gaussian beam 3 , the light sheet provides intrinsic optical sectioning and three dimensional imaging capabilities with significant background rejection. This approach, preventing the illumination of the whole sample volume, is particularly convenient for imaging of thick biological samples, such as embryos and whole brain 4-6 , since the imaging performances will strongly benefit of both the background reduction and the reduced phototoxicity. The unique features of light sheet fluorescence microscopy increase the penetration depth in the specimen and reduce the overall photobleaching. However, conventional implementations of selective plane illumination microscopes (SPIM) 7 require a suitable sample mounting, such as embedding the sample in a gel cylinder, thus preventing the possibility of imaging

Highly fluorescent (QDs)-loaded nanocapsules, intended for fluorescent cell imaging, were prepared via an emulsification/solvent-evaporation method. CdSe/ZnS core/shell quantum dots were applied as cargo, Poloxamer 403as the polymer component, Cremophor EL as the nonionic surfactant and mineral oil, oleic acid or silicone oil were applied as the oil phases. Transmission electron microscopy, atomic force microscopy, dynamic light scattering, and zeta potential measurements were used to characterize the novel QDs-labeled nanoparticles by particle size, distribution and morphology, as well as ζ-potential and physical stability. The fabricated long-lasting nanocapsules exhibit good luminescence properties upon both one-photon and twophoton excitation. The potential of the encapsulated QDs for fluorescent imaging was evaluated in cytotoxicity studies as well as in imaging of intracellular localization, accumulation and distribution of QDs delivered to well characterized human cancer cell lines -doxorubicinsensitive breast (MCF-7/WT) and alveolar basal epithelial (A549) -as well as on normal human umbilical vein endothelial (HUVEC) cells, performed by confocal laser scanning microscopy (CLSM). The colloidal CdSe/ZnS-loaded nanocapsules are shown to exhibit strong two-photon induced luminescence upon excitation in the NIR optical transmission window spectral range, making them ideal markers for bioimaging application. The total two-photon cross section of a single nanocapsule was determined to be about 4.1x10 6 GM at 800 nm.

Azine derivative 6,6'-((1E,1'E)-hydrazine-1,2-diylidenebis(methanyly-lidene))bis (3,4-dimethoxy-aniline) CHA has been synthesized by condensation of 6-nitroveratraldehyde with malononitrile followed by reduction using 10% Pd/C and hydrazine hydrate. In the presence of formaldehyde, CHA chemosensor in ethanol showed a yellow-to-red color change observable by the naked eyes and 'turn off' type fluorescence quenching in ethanol. This phenomenon was confirmed by UV-Vis, fluorescence and 1 H NMR spectroscopy. The CHA spectra in ethanol was shifted from 412 to 509 nm after addition of formaldehyde. Fluorescence intensity of CHA gradually decreased with the increase of formaldehyde concentrations. Furthermore, paper strips loaded-CHA were fabricated and served to detect formaldehyde qualitatively. The detection limit of the CHA probe to formaldehyde is 0.2 M.

Radiation based X-ray Fluorescence (SR nano-XRF) techniques were compared for a specific single cell imaging case study: high pressure frozen (HPF) and cryo-substituted polymorphonuclear neutrophils (PMNs). Nano-SIMS enabled nanoscale mapping of isotopes of C, N, O, P and S, while SR based nano-XRF enabled trace level imaging of metals like Ca, Mn, Fe, Ni, Cu and Zn at a resolution of approx. 50 nm. The obtained elemental distributions were compared with those of whole, cryofrozen PMNs measured at the newly developed ID16A nano-imaging beamline at the European Synchrotron Radiation Facility (ESRF) in Grenoble, France. Similarities were observed for elements more tightly bound to the cell structure such as phosphorus and sulphur, while differences for mobile ions such as chlorine and potassium were more pronounced. Due to the observed elemental redistribution of mobile ions such as potassium and chlorine, elemental analysis of high pressure frozen (HPF), cryo-substituted samples should therefore be interpreted critically. Although decreasing analytical sensitivity occurs due to the presence of ice, analysis of cryofrozen cells -close to their native state -remains the reference method par excellence. In general, we found nanoscale secondary ion mass spectrometry (nano-SIMS) and synchrotron radiation based nanoscopic X-ray fluorescence (SR nano-XRF) to be two supplementary alternatives, each with their own pros and cons, to investigate single cells at the nanoscale.

A series of complex boronic acids were prepared through multicomponent reactions (MCRs). Both Passerini and Ugi MCRs were carried out in which one component was an arylboronic acid. The resulting highly functionalized boronic acids participated efficiently in the Liebeskind-Srogl cross-coupling reaction with -methylthioBODIPY derivatives to yield complex borondipyrromethene (BODIPY) dyes in good yields. The joined spectroscopic and computational study points out the deep impact of the arylated chromophoric position on the photophysical signatures. Thus, unconstrained aryls grafted at the meso position did not sway the spectral band positions but switched on new nonradiative relaxation channels, whereas additional arylation at the opposite α-pyrrolic position softened such fluorescence quenching and shifted the emission to the red-edge of the visible spectrum. The conducted biological analysis revealed that peripheral blood mononuclear cells incubated with these new compounds showed ...

The design and optimization of fluorescent molecules has driven the ability to interrogate complex biological events in real time. Notably, most advances in bioimaging fluorophores are based on optimization of core structures that have been known for over a century. Recently, new synthetic methods have resulted in an explosion of nonplanar conjugated macrocyclic molecules with unique optical properties yet to be harnessed in a biological context. Herein we report the synthesis of the first aqueous-soluble carbon nanohoop (i.e., a macrocyclic slice of a carbon nanotube prepared via organic synthesis) and demonstrate its bioimaging capabilities in live cells. Moreover, we illustrate that these scaffolds can be easily modified by well-established "click" chemistry to enable targeted live cell imaging. This work establishes the nanohoops as an exciting new class of macrocyclic fluorophores poised for further development as novel bioimaging tools.

In the twenty-first century, nanotechnology has become cutting-edge technology. It is interdisciplinary and multidisciplinary, covering numerous fields such as medicine, engineering, biology, physics, material sciences, and chemistry. The present work aims to cover the optical properties, method of preparations, surface modifications, bio-conjugation, characterization, stability, and cytotoxicity of quantum dots (QDs). Articles were reviewed in English literature reporting the pharmaceutical and bio-pharmaceutical aspects of QDs which were indexed in Scopus, web of science, google scholar and PubMed without applying the year of publication criterion. One significant value of utilizing nanotechnology is that one can alter and control the properties in a genuinely unsurprising way to address explicit applications' issues. In science and biomedicine, the usage of functional nanomaterials has been broadly investigated and has become one of the quick-moving and stimulating research d...

A series of complex boronic acids were prepared through multicomponent reactions (MCRs). Both Passerini and Ugi MCRs were carried out in which one component was an arylboronic acid. The resulting highly functionalized boronic acids participated efficiently in the Liebeskind-Srogl cross-coupling reaction with -methylthioBODIPY derivatives to yield complex borondipyrromethene (BODIPY) dyes in good yields. The joined spectroscopic and computational study points out the deep impact of the arylated chromophoric position on the photophysical signatures. Thus, unconstrained aryls grafted at the meso position did not sway the spectral band positions but switched on new nonradiative relaxation channels, whereas additional arylation at the opposite α-pyrrolic position softened such fluorescence quenching and shifted the emission to the red-edge of the visible spectrum. The conducted biological analysis revealed that peripheral blood mononuclear cells incubated with these new compounds showed ...

Structured illumination microscopy (SIM) with axially optical sectioning capability has found widespread applications in three-dimensional live cell imaging in recent years, since it combines high sensitivity, short image acquisition time, and high spatial resolution. To obtain one sectioned slice, three raw images with a fixed phase-shift, normally 2π/3, are generally required. In this paper, we report a data processing algorithm based on the one-dimensional Hilbert transform, which needs only two raw images with arbitrary phase-shift for each single slice. The proposed algorithm is different from the previous two-dimensional Hilbert spiral transform algorithm in theory. The presented algorithm has the advantages of simpler data processing procedure, faster computation speed and better reconstructed image quality. The validity of the scheme is verified by imaging biological samples in our developed DMD-based LED-illumination SIM system.

Somatic embryogenesis (SE) is one of the most promising processes for large-scale dissemination of elite varieties. However, for many plant species, optimizing SE protocols still relies on a trial-and-error approach. Using coffee as a model plant, we report here the first global analysis of metabolome and hormone dynamics aiming to unravel mechanisms regulating cell fate and totipotency. Sampling from leaf explant dedifferentiation until embryo development covered 15 key stages. An in-depth statistical analysis performed on 104 metabolites revealed that massive re-configuration of metabolic pathways induced SE. During initial dedifferentiation, a sharp decrease in phenolic compounds and caffeine levels was also observed while auxins, cytokinins and ethylene levels were at their highest. Totipotency reached its highest expression during the callus stages when a shut-off in hormonal and metabolic pathways related to sugar and energetic substance hydrolysis was evidenced. Abscisic acid...

In this paper, we describe both new as well as described in our previous works 1,8naphthalimide derivatives substituted at the 3-C position with imine or -ketoenamine unitin order to demonstrate a broader scope of research enabling of analysis between the structureproperties relationship relevant to the application of these compounds in organic electronics and cellular imaging. Thermal, physicochemical, optical, electrochemical, electroluminescence, and biological properties of a series of derivatives containing the 1,8-naphthalimideunit were tested and compared. This allowed the determination of impact of substituents in the imide part (hexylamine, phenylethyl, benzyl, fluorobenzyl, methylbenzyl), type of bond (imine or ketoenamine) as well as the substituent on the naphthalene ring (2-hydroxyphenyl, 5-bromo-2hydroxyphenyl, 3,5-diodo-2-hydroxyphenyl, pyrimidines) on their properties. Moreover, the properties in the aggregating state were tested in the MeOH/PBS system. Imines are susceptible to the hydrolysis process and aggregation-caused photoluminescence quenching(ACQ). In turn,-ketoenamine shown excited-state intramolecular proton transfer promoted by aggregation (AIEE). Our studies can be helpful in the further design of compounds containing the 1,8naphthalimide structure for various applications.

A new series of 1,8-naphtalimides containing an imine bond at the 3-position of the naphthalene ring was synthesized using 1H, 13C NMR, FTIR, and elementary analysis. The impact of the substituent in the imine linkage on the selected properties and bioimaging of the synthesized compounds was studied. They showed a melting temperature in the range of 120–164 °C and underwent thermal decomposition above 280 °C. Based on cyclic and differential pulse voltammetry, the electrochemical behavior of 1,8-naphtalimide derivatives was evaluated. The electrochemical reduction and oxidation processes were observed. The compounds were characterized by a low energy band gap (below 2.60 eV). Their photoluminescence activities were investigated in solution considering the solvent effect, in the aggregated and thin film, and a mixture of poly(N-vinylcarbazole) (PVK) and 2-tert-butylphenyl-5-biphenyl-1,3,4-oxadiazole (PBD) (50:50 wt.%). They demonstrated low emissions due to photoinduced electron tran...

In this paper, we describe both new as well as described in our previous works 1,8naphthalimide derivatives substituted at the 3-C position with imine or -ketoenamine unitin order to demonstrate a broader scope of research enabling of analysis between the structureproperties relationship relevant to the application of these compounds in organic electronics and cellular imaging. Thermal, physicochemical, optical, electrochemical, electroluminescence, and biological properties of a series of derivatives containing the 1,8-naphthalimideunit were tested and compared. This allowed the determination of impact of substituents in the imide part (hexylamine, phenylethyl, benzyl, fluorobenzyl, methylbenzyl), type of bond (imine or ketoenamine) as well as the substituent on the naphthalene ring (2-hydroxyphenyl, 5-bromo-2hydroxyphenyl, 3,5-diodo-2-hydroxyphenyl, pyrimidines) on their properties. Moreover, the properties in the aggregating state were tested in the MeOH/PBS system. Imines are susceptible to the hydrolysis process and aggregation-caused photoluminescence quenching(ACQ). In turn,-ketoenamine shown excited-state intramolecular proton transfer promoted by aggregation (AIEE). Our studies can be helpful in the further design of compounds containing the 1,8naphthalimide structure for various applications.

Objective: To provide information about the phytochemical features of the crude extract (CruE) and fractions of the forageBauhinia glabra Jacq., Fabaceae, and present themultivariate correlation between its metabolites and the antioxidant properties. Methods: Studies were carried out by extraction with ethanol, sequential partition with hexane, chloroform and ethyl acetate, determination of the total yield of extraction, qualitative and quantitative estimation of phytochemicals.Then, the evaluation of antioxidant properties by four methods: 1,1-diphenyl picrylhydrazyl free radical scavenging assay (DPPH), the reduction power, the evaluation of inhibition of lipid peroxidation (TBARS) and the evaluation of recovery of content of methemoglobin in erythrocytes. Results: Phytochemical analysis on CruErevealed the presence of chlorophylls, carotenoids, coumarins, phytosterols, anthocyanins, tannins and flavonoids. Quantitative estimation of metabolites on CruE showed high concentration of phytosterols (42.21±2.34) mg/gdw and total phenolic (58.50±1.98) mg/gdw. The best results onthe antioxidant properties on each assay were the chloroform fraction (ChlF) with 13.76±0.23 and methanolic fraction (MetF) with 13.46±0.45 of half-maximal inhibitory concentration (IC50 Conclusion: Based on multivariate analysis, HexFthat presented the highest concentration of phytosterols, has the highest reduction power and was more effectivefor inhibition of lipid peroxidation and in recoveringmethemoglobincontent. Therefore, the mechanism of these activities seems to be related to the reduction power presented by phytosterols.) values (DPPH).The HexF(60.99±1.76%) and ChlF(64.40±1.73%), both based on reduction power of ascorbic acid. The HexF(73.54±2.74%) of inhibition of lipid peroxidation (TBARS), and HexFwith 7.28±0.36% amount of methemoglobin.

Cost-effective anti-cancer delivery vehicles that can ensure controlled and targeted transportation of drug molecules are pertinent to modern day biomedical applications. Minimally toxic 9-13 nm diameter porous carbon nanospheres (PNs) were synthesized by oxidative cutting of porous carbon matrices (PCs) obtained by carbonization of pasture grass, human hair and sucrose. Among them, the grass-derived PNs (PN-G) with superior surface area, porosity and graphitic content demonstrates a significant loading of the drug both by chemical binding and physisorption. Polyethylenimine (PEI) and folic acid (FA) functionalization maintain therapeutic efficacy of the drug doxorubicin (DOX) to the targeted folate receptor (FR) over-expressed human cervical cancer cells (HeLa) and human breast cancer cells (MDA-MB-231) through receptor mediated endocytosis while FR deficient normal cells (human embryonic kidney 293) exhibits substantially lower endocytosis under identical conditions. Moreover, upon loading cellimpermeable propidium iodide (PI), the PNs display superior activity towards near infrared (NIR) live cell imaging in HeLa cells whereby due to a higher binding affinity of PI with the nucleic acids, the PI-to-PN energy transfer quenched fluorescence is recovered. This dual functionality of controlled and targeted drug delivery and photobleaching resistant live cell imaging by the cost-effective PNs has larger implications in nanomedicine research and technology.

Since the great achievement and progress made for the generation of novel nanostructures, 2 theranostic nano-platforms have been the trend topic due to their intensive capability of therapy and diagnosis. Hence, theranostics have also been a generic strategy for the personalized 4 medicine, recently. Moreover, traditional therapy modalities limit the use of chemotherapeutic 5 agents for every patient and this requires more effective drug carrier systems by designing the 6 formulation of drug in a specified way. Herein, we performed a generic theranostic platform in 7 "all-in-one" concept by the combination of two therapy modalities with active targeting 8 approach. To achieve this, 10 nm gold nanoparticles (AuNPs) and protoporphyrine IX (PpIX) 9 were encapsulated into folic acid (FA) tagged niosome vesicles. Resulted AuNP-PpIX-FA 10 niosomes were characterized and their particle size was 93±17 nm with a high surface charge 11 and encapsulation efficiency (around 85%). In the case of bio-applications for AuNP-PpIX-FA 12 niosomes, folate receptor positive (FR(+)) human cervical cancer cell line (HeLa) and FR 13 negative (FR(-)) human alveolar type-II (A549) like cell line were examined with the relative 14 control groups of theranostic vesicles. By testing the toxicity of vesicles, non-toxic 15 concentrations were introduced to cell with the combined treatment of radiotherapy and 16 photodynamic therapy, successfully. On the other hand, cellular uptake of niosomes were also 17 showed its great potential for FR (+) HeLa cells as the theranostic platform with all-in-one 18 approach. 19 20

Since the great achievement and progress made for the generation of novel nanostructures, 2 theranostic nano-platforms have been the trend topic due to their intensive capability of therapy and diagnosis. Hence, theranostics have also been a generic strategy for the personalized 4 medicine, recently. Moreover, traditional therapy modalities limit the use of chemotherapeutic 5 agents for every patient and this requires more effective drug carrier systems by designing the 6 formulation of drug in a specified way. Herein, we performed a generic theranostic platform in 7 "all-in-one" concept by the combination of two therapy modalities with active targeting 8 approach. To achieve this, 10 nm gold nanoparticles (AuNPs) and protoporphyrine IX (PpIX) 9 were encapsulated into folic acid (FA) tagged niosome vesicles. Resulted AuNP-PpIX-FA 10 niosomes were characterized and their particle size was 93±17 nm with a high surface charge 11 and encapsulation efficiency (around 85%). In the case of bio-applications for AuNP-PpIX-FA 12 niosomes, folate receptor positive (FR(+)) human cervical cancer cell line (HeLa) and FR 13 negative (FR(-)) human alveolar type-II (A549) like cell line were examined with the relative 14 control groups of theranostic vesicles. By testing the toxicity of vesicles, non-toxic 15 concentrations were introduced to cell with the combined treatment of radiotherapy and 16 photodynamic therapy, successfully. On the other hand, cellular uptake of niosomes were also 17 showed its great potential for FR (+) HeLa cells as the theranostic platform with all-in-one 18 approach. 19 20

Structured illumination microscopy (SIM) with axially optical sectioning capability has found widespread applications in three-dimensional live cell imaging in recent years, since it combines high sensitivity, short image acquisition time, and high spatial resolution. To obtain one sectioned slice, three raw images with a fixed phase-shift, normally 2π/3, are generally required. In this paper, we report a data processing algorithm based on the one-dimensional Hilbert transform, which needs only two raw images with arbitrary phase-shift for each single slice. The proposed algorithm is different from the previous two-dimensional Hilbert spiral transform algorithm in theory. The presented algorithm has the advantages of simpler data processing procedure, faster computation speed and better reconstructed image quality. The validity of the scheme is verified by imaging biological samples in our developed DMD-based LED-illumination SIM system.

Somatic embryogenesis (SE) is one of the most promising processes for large-scale dissemination of elite varieties. However, for many plant species, optimizing SE protocols still relies on a trial-and-error approach. Using coffee as a model plant, we report here the first global analysis of metabolome and hormone dynamics aiming to unravel mechanisms regulating cell fate and totipotency. Sampling from leaf explant dedifferentiation until embryo development covered 15 key stages. An in-depth statistical analysis performed on 104 metabolites revealed that massive re-configuration of metabolic pathways induced SE. During initial dedifferentiation, a sharp decrease in phenolic compounds and caffeine levels was also observed while auxins, cytokinins and ethylene levels were at their highest. Totipotency reached its highest expression during the callus stages when a shut-off in hormonal and metabolic pathways related to sugar and energetic substance hydrolysis was evidenced. Abscisic acid...

Cell classification refers to detecting normal and diseased cells from small amount of data. Sometimes, classification of cells becomes difficult because some cells fall into more than one categories/classes. Current state‐of‐the‐art cell classification methods have been developed on the bases of tumor cell classification but these methods cannot classify diseased or normal cells. This study investigated the performance of two classification methods traditional machine learning and deep learning (normal and diseased cell classification) to categorize normal and diseased cells. Millions of normal cells undergo controlled growth and uncontrolled growth may be involved in disease causation but their clinical applications remain limited due to difficulties in distinguishing normal and diseased cells. Previous studies are limited to identify, systematically, the normal and diseased cells. This study collected information about diseased or normal cells to check the networks for correct ce...

Fluorescent semiconductor nanoparticles in tree-dimensional quantum confinement, quantum dots (QDs), synthesized in aqueous medium, and functionalized with polyethylene glycol, were used as nonspecific nanosized probes for the long-term imaging of glial cells. In vitro living healthy as well as cancer glial cells were labelled by direct insertion of a small volume of QDs contained in aqueous suspension into the culture wells. A long-term monitoring (over 7 days) of the cells was performed and no evidence of cell fixation and/or damage was observed. Two control groups, healthy and non-healthy glial cells, were used to compare cell viability. During the observation period, labelled cells kept the same behavior compared to non-labelled control samples. To our knowledge, this is the first report of the viability of quantum dots for long-term imaging of living cells. This opens a large range of possibilities related to a better understand of cellular mechanisms, which till now was not ac...

학위논문(박사)--서울대학교 대학원 :공과대학 기계공학부,2019. 8. Lee, Junghoon.Commercial high-resolution optical microscopes are essential for microscopy imaging; however, they are expensive and bulky, which limits their use in point-of-care devices, resource-limited areas, and real-time imaging of a sample in a large apparatus. In this study, we report a novel compact lightweight submicron-resolution reflected and inverted optical microscope at low cost. Our technique utilizes the proximity of the image sensor to a commercial microscope objective lens for compactness of the microscope. The use of an image sensor with a small pixel size helps to reduce the information loss, which provides high-resolution images. Moreover, our technique offers a freedom to tailor the design of microscope according to the required resolution, cost, and portability for specific applications, which makes it a suitable candidate for affordable point-of-care devices. Unlike conventional reflected microscope, which consists of d...

An organic nanoparticle-based drug delivery system with high drug loading efficacy (∼79 wt %) was developed using a perylene-derived photoremovable protecting group, namely, perylene-3,4,9,10-tetrayltetramethanol (Pe(OH) 4). The anticancer drug chlorambucil was protected by coupling with Pe(OH) 4 to form photocaged nanoparticles (Pe(Cbl) 4). The photorelease mechanism of chlorambucil from the Pe(Cbl) 4 conjugate was investigated experimentally by high-resolution mass spectrometry and theoretically by density functional theory calculations. The Pe(Cbl) 4 nanoparticles perform four important roles: (i) a nanocarrier for drug delivery, (ii) a phototrigger for drug release, (iii) a fluorescent chromophore for cell imaging, and (iv) a photoswitchable fluorophore for real-time monitoring of drug release. Tunable emission of the perylene-derived nanoparticles was demonstrated by comparing the emission properties of the Pe(OH) 4 and Pe(Cbl) 4 nanoparticles with perylene-3-ylmethanol. These nanoparticles were subsequently employed in cell imaging for investigating their intracellular localization. Furthermore, the in vivo toxicity of the Pe(OH) 4 nanoparticles was investigated using the mouse model. Histological tissue analysis of five major organs, i.e., heart, kidney, spleen, liver, and lung, indicates that the nanoparticles did not show any obvious damage to these major organs under the experimental conditions. The current research presents a successful example of integrating multiple functions into single-component organic nanoparticles for drug delivery.

In the twenty-first century, nanotechnology has become cutting-edge technology. It is interdisciplinary and multidisciplinary, covering numerous fields such as medicine, engineering, biology, physics, material sciences, and chemistry. The present work aims to cover the optical properties, method of preparations, surface modifications, bio-conjugation, characterization, stability, and cytotoxicity of quantum dots (QDs). Articles were reviewed in English literature reporting the pharmaceutical and bio-pharmaceutical aspects of QDs which were indexed in Scopus, web of science, google scholar and PubMed without applying the year of publication criterion. One significant value of utilizing nanotechnology is that one can alter and control the properties in a genuinely unsurprising way to address explicit applications' issues. In science and biomedicine, the usage of functional nanomaterials has been broadly investigated and has become one of the quick-moving and stimulating research d...

Cell rotation reveals important information which facilitates identification and characterization of different cells. Markedly, achieving three dimensional (3D) rolling rotation of single cells within a larger group of cells is rare among existing cell rotation techniques. In this work we present a simple biochip which can be used to trap and rotate a single cell, or to rotate multiple cells relative to each other within a group of individual red blood cells (RBCs), which is crucial for imaging cells in 3D. To achieve single RBC trapping, we employ two parallel sidewall 3D electrodes to produce a dielectrophoretic force which traps cells inside the capturing chambers of the microfluidic device, where the hydrodynamic force then induces precise rotation of the cell inside the chamber. We have also demonstrated the possibility of using the developed biochip to preconcentrate and rotate RBC clusters in 3D. As our proposed cell trapping and rotation device reduces the intricacy of cell rotation, the developed technique may have important implications for high resolution 3D cell imaging in the investigation of complex cell dynamics and interactions in moving media.

The development of novel uncomplicated sensor for sensitive and selective detection in biomedical analysis is necessary. This study demonstrates the preparation of nitrogen doped carbon dots (N-C-dots) via hydrothermal synthesis as a naked eye fluorescent probe for selective and sensitive detection of dopamine (DA). The successful synthesis of N-C-dots was confirmed using various microscopic and spectroscopic techniques. In addition, N-C-dots incorporated gel strips were prepared through a simple approach for naked eye detection of dopamine. The linear range for detection of dopamine was 2 to 20 µg/mL with a detection limit as low as 1.97 µg/mL. Moreover N-C-dots showed low toxicity towards HeLa cells demonstrating their relevance in multicolour cell imaging. The developed probe was also utilized for quantitative determination of dopamine in commercial pharmaceutical sample with well adequate results.

Most multicellular organisms require apoptosis, or programmed cell death, to function properly and survive. On the other hand, morphological and biochemical characteristics of apoptosis have remained remarkably consistent throughout evolution. Apoptosis is thought to have at least three functionally distinct phases: induction, effector, and execution. Recent studies have revealed that reactive oxygen species (ROS) and the oxidative stress could play an essential role in apoptosis. Advanced microscopic imaging techniques allow biologists to acquire an extensive amount of cell images within a matter of minutes which rule out the manual analysis of image data acquisition. The segmentation of cell images is often considered the cornerstone and central problem for image analysis. Currently, the issue of segmentation of mitochondrial cell images via deep learning receives increasing attention. The manual labeling of cell images is time-consuming and challenging to train a pro. As a courte...

In the twenty-first century, nanotechnology has become cutting-edge technology. It is interdisciplinary and multidisciplinary, covering numerous fields such as medicine, engineering, biology, physics, material sciences, and chemistry. The present work aims to cover the optical properties, method of preparations, surface modifications, bio-conjugation, characterization, stability, and cytotoxicity of quantum dots (QDs). Articles were reviewed in English literature reporting the pharmaceutical and bio-pharmaceutical aspects of QDs which were indexed in Scopus, web of science, google scholar and PubMed without applying the year of publication criterion. One significant value of utilizing nanotechnology is that one can alter and control the properties in a genuinely unsurprising way to address explicit applications' issues. In science and biomedicine, the usage of functional nanomaterials has been broadly investigated and has become one of the quick-moving and stimulating research d...

Supplemental Figure 9-Representative high performance liquid chromatography profile of Momordica charantia extract. Gallic acid (peak 1), catechin (peak 2), chlorogenic acid (peak 3), caffeic acid (peak 4), ellagic acid (peak 5), epicatechin (peak 6), rutin (peak 7), quercitrin (peak 8), isoquercitrin (peak 9), quercetin (peak 10) and kaempferol (peak 11). Supplementary Figure 8-Representative high performance liquid chromatography profile of Phyllanthus Amarus extract. Gallic acid (peak 1), catechin (peak 2), chlorogenic acid (peak 3), ellagic acid (peak 4), epicatechin (peak 5), rutin (peak 6), quercitrin (peak 7), isoquercitrin (peak 8), quercetin (peak 9) and kaempferol (peak 10).

Three novel and facile calixarene derivatives (5, 6 and 7), which were appended with four rhodamine units at the upper rim of calixarene skeleton, were firstly prepared and evaluated for selective detection of metal ions in solution. Receptors (5) and (7) indicated immediate turn on fluorescence output toward Hg 2+ ions over other most competitive metal ions with the ultralow detection limits, indicating their high efficiency and reliability. The binding response to Hg 2+ ions in solution was also observed through a chromogenic change (from colorless to pale pink). Furthermore, in vitro and bio-imaging studies with MCF-7 or MIA PaCa-2 cell lines were also performed to investigate the use of receptors in biological systems in order to monitor of mercury ions. Results showed that new receptors (5) or (7) were cell permeable and suitable for real-time imaging of Hg 2+ in living cells (MCF-7) or (MIA PaCa-2) without any damage to healthy cell lines (HEK 293).

A facile synthesis procedure, whereby 9-Anthraldehyde (AA) is coupled to aminated rhodamine (AR) via a Schiff base-type reaction, is reported. The applicability and performance of the obtained material (AA-AR) as a sensing agent was studied towards 16 metal cations (i.

In the twenty-first century, nanotechnology has become cutting-edge technology. It is interdisciplinary and multidisciplinary, covering numerous fields such as medicine, engineering, biology, physics, material sciences, and chemistry. The present work aims to cover the optical properties, method of preparations, surface modifications, bio-conjugation, characterization, stability, and cytotoxicity of quantum dots (QDs). Articles were reviewed in English literature reporting the pharmaceutical and bio-pharmaceutical aspects of QDs which were indexed in Scopus, web of science, google scholar and PubMed without applying the year of publication criterion. One significant value of utilizing nanotechnology is that one can alter and control the properties in a genuinely unsurprising way to address explicit applications' issues. In science and biomedicine, the usage of functional nanomaterials has been broadly investigated and has become one of the quick-moving and stimulating research directions. Different types of nanomaterial (silicon nanowires, QDs, carbon nanotubes, nanoparticles of gold/silver) were extensively utilized for biological purposes. Nanomedicine shows numerous advantages in the natural characteristics of targeted drug delivery and therapeutics. For instance, protection of drugs against degradation, improvement in the drug's stability, prolonged circulation time, deceased side effects, and enhanced distribution in tissues. The present review article deals with the quantum dots, their optical properties, method of preparations, surface modifications, bio-conjugation, characterization, stability, and cytotoxicity of quantum dots. The review also discusses various biomedical applications of QDs. The QDs-based bio-nanotechnology will always be in the growing list of unique applications, with progress being made in specialized nanoparticle development, the detection of elegant conjugation methods, and the discovery of new targeting ligands.

Colon cancer is one of the world's most deadly diseases. Because of its internal location, it is necessary to obtain faster and more efficient diagnostic tools for this organ site. In this context, we studied the development of new luminescent nanoprobes (LNPs) as an alternative diagnostic apparatus for detecting this disease. The nanoparticles examined herein are lanthanide-doped sodium yttrium fluoride (NaYF 4 :Ln) and have shown to be promising as investigative devices. However, significant problems with the use of LNPs are the lack of biocompatibility and the targeting of the system to tumor regions. One of the strategies to bypass these problems is to increase of the particle lipophilicity modifying their surfaces with organic compounds that present high similarity to the biological system. In this work, we synthesized six new materials for use in bioimaging techniques obtained from the combination of nanoparticles of NaYF 4 :5%Eu with organic aromatic compounds covalently bonded. The materials were characterized structurally and morphologically using XRD and TEM, techniques, which showed the identification of the crystallographic phase β-NaYF 4 :5%Eu and its nanometric size (particles smaller than 50 nm). The conjugation process was confirmed by FT-IR spectra analysis and from the TGA profile. Excitation and emission spectra allowed the evaluation of the optical properties of the synthesized compounds. The interaction and cellular uptake was confirmed when HT-29 colon cancer cells were exposed to LNPs, indicating that the developed system has promising applications in bioimaging procedures.

The present article delineates the formation of green fluorescent organic nanoparticle through supramolecular aggregation of naphthalene diimide (NDI) based carboxybenzyl protected L-phenylalanine appended bola-amphiphile, NDI-1. The amphiphilic molecule is soluble in DMSO and with gradual addition of water within the DMSO solution, the amphiphile starts to get self-assembled via H-type aggregation to form spherical nanoparticles. These self-assembly of NDI-1 in presence of high amount of water, exhibited aggregation induced emission (AIE) through excimer formation. Notably, in presence of 99% water content the amphiphile forms spherical aggregated nanoparticles as confirmed from microscopic investigations and dynamic light scattering study. Interestingly, the emission maxima of molecularly dissolved NDI-1 (weak blue fluorescence) gets red shifted upon aggregation with increase in water concentration and led to the formation of green emitting fluorescent organic nanoparticles (FONPs...

9-Acridone-4-carboxylic acid has been established as a cheap, non-carcinogenic fluorescence dye. It can differentiate living and dead cells in a eukaryotic microbial population viz. Candida sp. Moreover, the dye has been tagged with specific antibody (Ab) without compromising its fluorescence property and used for detection of specific antigen in natural samples. The dye crystallizes in the monoclinic space group C2/c with unit cell dimensions : a = 17.7608(10) Å, b = 7.3009(4) Å and c = 16.7328(9) Å while other unit cell parameters are :  = 90º,  = 103.093(4)º,  = 90º, V = 2113.3(2) Å 3 and Z = 8. The dye is soluble in HEPES buffer at pH 7.4.

Triplaris gardneriana Wedd. is a tree used in folk medicine to treat venereal diseases and inflammation as well as a source of biological compounds with antioxidant capacity. In order to assess the safety of these bioactive compounds, the present study aimed to determine the toxicity of an ethanolic extract of T. gardneriana, (EETg). Toxicological tests included hemolytic activity, toxicity toward the brine shrimp Artemia, cytotoxicity against breast cancer cells (MCF7) and acute oral toxicity in rodents. In addition, toxicogenomics techniques were used to determine genome expression in MCF7 cells exposed to EETg. The results showed that the extract exhibits approximately 60% of hemolytic activity at the highest tested concentration (64 µg/ml) and toxicity against nauplii of Artemia sp. (LC 50 of 67.85 µg/ml). Further, EETg appears to be cytotoxic to MCF7 (cell viability reduced to 40% at 250 µg/ml after 24 hr). Genomic data demonstrated differential expression of 14 genes. Data analysis indicated possible altered pathways (e.g., xenobiotic metabolism), possible adverse health risks (e.g., hepatotoxicity), and drugs with similar gene expression profile (e.g., antimicrobials). The investigation provides important information on potentially adverse aspects of EETg, which need to be considered prior to the therapeutic utilization of this plant. Abbreviations: EETg: ethanolic extract of T. gardneriana seeds; MCF7: michigan cancer foundation-7 which refers to a human breast cell line (adenocarcinoma); NGS: next-generation sequencing; edgeR: empirical analysis of digital gene expression data in R; Consensus: consensus path database; FDR: false discovery rate; NCBI: national center for biotechnology information; KEGG: kyoto encyclopedia of genes and genomes; Ingenuity: ingenuity pathway analysis software; CMAP: connectivity map; OECD: organization for economic cooperation and development; HL-60: human promyelocytic leukemia cells; PC3: prostate cancer cells.

Cell classification refers to detecting normal and diseased cells from small amount of data. Sometimes, classification of cells becomes difficult because some cells fall into more than one categories/classes. Current state-of-the-art cell classification methods have been developed on the bases of tumor cell classification but these methods cannot classify diseased or normal cells. This study investigated the performance of two classification methods traditional machine learning and deep learning (normal and diseased cell classification) to categorize normal and diseased cells. Millions of normal cells undergo controlled growth and uncontrolled growth may be involved in disease causation but their clinical applications remain limited due to difficulties in distinguishing normal and diseased cells. Previous studies are limited to identify, systematically, the normal and diseased cells. This study collected information about diseased or normal cells to check the networks for correct cell detection and then eliminated false-positive cells. We used machine learning methods along with logistic regression, support vector machine, and CNN (convolutional neural network). We found that our proposed method classified better the normal and diseased cells. With the help of two types of images: normal and diseased cells, we trained a CNN that identified diseased cells with 98% accuracy and enabled the discovery of normal and diseased cells. As a result, it will advance the clinical utility of human diseased cells.

Structured illumination microscopy (SIM) with axially optical sectioning capability has found widespread applications in three-dimensional live cell imaging in recent years, since it combines high sensitivity, short image acquisition time, and high spatial resolution. To obtain one sectioned slice, three raw images with a fixed phase-shift, normally 2π/3, are generally required. In this paper, we report a data processing algorithm based on the one-dimensional Hilbert transform, which needs only two raw images with arbitrary phase-shift for each single slice. The proposed algorithm is different from the previous two-dimensional Hilbert spiral transform algorithm in theory. The presented algorithm has the advantages of simpler data processing procedure, faster computation speed and better reconstructed image quality. The validity of the scheme is verified by imaging biological samples in our developed DMD-based LED-illumination SIM system.

Cell classification refers to detecting normal and diseased cells from small amount of data. Sometimes, classification of cells becomes difficult because some cells fall into more than one categories/classes. Current state-of-the-art cell classification methods have been developed on the bases of tumor cell classification but these methods cannot classify diseased or normal cells. This study investigated the performance of two classification methods traditional machine learning and deep learning (normal and diseased cell classification) to categorize normal and diseased cells. Millions of normal cells undergo controlled growth and uncontrolled growth may be involved in disease causation but their clinical applications remain limited due to difficulties in distinguishing normal and diseased cells. Previous studies are limited to identify, systematically, the normal and diseased cells. This study collected information about diseased or normal cells to check the networks for correct cell detection and then eliminated false-positive cells. We used machine learning methods along with logistic regression, support vector machine, and CNN (convolutional neural network). We found that our proposed method classified better the normal and diseased cells. With the help of two types of images: normal and diseased cells, we trained a CNN that identified diseased cells with 98% accuracy and enabled the discovery of normal and diseased cells. As a result, it will advance the clinical utility of human diseased cells.

Multifaceted application potential of AEEgens in bioimaging, theranostics, chemo/biosensors, mechanochromics, solar cells, and organic photoelectronics opens up a new research paradigm to develop and design more such compounds. Herein, quinoxaline N-directed Ru(II)-catalyzed oxidative annulation of 2-arylquinoxalines with internal alkynes leads to the formation of highly luminescent annulated quaternary ammonium salts in the presence of a Cu(OAc) 2 •H 2 O oxidant. While the synthesized compounds exhibit emissions in the green-to-yellow region with large Stokes shifts and reasonable quantum yields, their DFT calculations display a 3D twisted conformation bearing a donor−π−acceptor (D-π-A) configuration where the two phenyl moieties could serve as the donors and the extended quinoxaline core as the acceptor. Single-crystal analysis of the quaternary salt 3aa depicts the presence of multiple intermolecular noncovalent and weak π−π interactions that are possibly responsible for the luminescence behavior in crystalline and solid states. The advent of aggregation-enhanced emission in quinoxalinium salt 3aa in DMF/water is due to the restriction of intramolecular motion and suppression of intermolecular charge transfer in the aggregated state. AEEgen 3aa unveils reversible mechanochromism on changing from crystalline to the amorphous state upon grinding and returning back to the crystalline state upon DCM fuming, where a few of such compounds are utilized for development of latent fingerprints on adhesive tape. Furthermore, a representative group of synthesized luminescent quinoxalinium salts portrays dose-dependent cell growth inhibition of HeLa cells with concomitant cell arrest in G1 phases. Hence, these AEE luminogens are not only attractive as luminescent "light-up" probes for cell imaging but also important as anticancer agents.

Nonlinear microscopes have seen an increase in popularity in the life sciences due to their molecular and structural specificity, high resolution, large penetration depth, and volumetric imaging capability. Nonetheless, the inherently weak optical signals demand long exposure times for live cell imaging. Here, by modifying the optical layout and illumination parameters, we can follow the rotation and translation of noncentrosymetric crystalline particles, or nanodoublers, with 50 μs acquisition times in living cells. The rotational diffusion can be derived from variations in the second harmonic intensity that originates from the rotation of the nanodoubler crystal axis. We envisage that by capitalizing on the biocompatibility, functionalizability, stability, and nondestructive optical response of the nanodoublers, novel insights on cellular dynamics are within reach.

Scanning transmission ion microscopy (STIM) imaging of living cultured cells has been carried out using a proton external-beam with the nuclear microprobe of Bordeaux-Gradignan. STIM could be performed in air atmosphere after passage of a focused proton beam through a 150 nm thick silicon nitride window. Energy loss STIM images were obtained with a spatial resolution in the micrometer range and enabled the identi®cation of sub-cellular ultrastructures.