Multiplexed stimulated emission depletion nanoscopy(mSTED)for 5-color live-cell long-term imaging of organelle interactome

Stimulated emission depletion microscopy(STED)holds great potential in biological science applications,especially in studying nanoscale subcellular structures.However,multi-color STED imaging in live-cell remains challenging due to the limited excitation wavelengths and large amount of laser radiation.Here,we develop a multiplexed live-cell STED method to observe more structures simultaneously with limited photo-bleaching and photo-cytotoxicity.By separating live-cell fluorescent probes with similar spectral properties using phasor analysis,our method enables five-color live-cell STED imaging and reveals long-term interactions between different subcellular structures.The results here provide an avenue for understanding the complex and delicate interactome of subcellular structures in live-cell.

Fluorescence imaging analysis of the glioma microenvironment

Glioma is the most malignant brain cancer.The neurons,macrophages,T cells and other immune ellls constitute the glioma immunosuppressive microenvironment.The accurate spatial distri-bution of these cells in the glioma microenvironment and its relationship with glioma metastasis is unknown.We constructed a mouse gliomna cell line stably expressing the large Stokes shifted yellow fluorescent protein and applied it to the multicolor immunofluorescence imaging.The inaging data revealed that the neurons were sparsely distributed in the glioma core and the mumber of neurons decreased by 90%compared with normal brain site.The spatial distribution of monocyte-macrophages and microglia is heterogeneous.The monocyte macrophages and T cells were heavily recrnuited into the glioma core and metastasis.There was no significant difference in the distribution of microglia amnong glioma core,margin,and normal brain site.Our results provided new perspectives for targeting immune regulation cells and developing new immuno-therapy strategies for glioma.

Intestinal natural killer/T-cell lymphoma presenting as a pancreatic head space-occupying lesion:A case report

BACKGROUND Intestinal natural killer/T-cell lymphoma(NKTCL)is a rare and aggressive non-Hodgkin’s lymphoma,and its occurrence is closely related to Epstein-Barr virus infection.In addition,the clinical symptoms of NKTCL are not obvious,and the specific pathogenesis is still uncertain.While NKTCL may occur in any segment of the intestinal tract,its distinct location in the periampullary region,which leads clinicians to consider mimics of a pancreatic head mass,should also be addressed.Therefore,there remain huge challenges in the diagnosis and treatment of intestinal NKTCL.CASE SUMMARY In this case,we introduce a male who presented to the clinic with edema of both lower limbs,accompanied by diarrhea,and abdominal pain.Endoscopic ultrasound(EUS)showed well-defined homogeneous hypoechoic lesions with abundant blood flow signals and compression signs in the head of the pancreas.Under the guidance of EUS-fine needle biopsy(FNB)with 19 gauge or 22 gauge needles,combined with multicolor flow cytometry immunophenotyping(MFCI)helped us diagnose NKTCL.During treatments,the patient was prescribed the steroid(dexamethasone),methotrexate,ifosfamide,L-asparaginase,and etoposide chemotherapy regimen.Unfortunately,he died of leukopenia and severe septic shock in a local hospital.CONCLUSION Clinicians should enhance their understanding of NKTCL.Some key factors,including EUS characteristics,the right choice of FNB needle,and combination with MFCI,are crucial for improving the diagnostic rate and reducing the misdiagnosis rate.

Chromosome analysis of esophageal squamous cell carcinoma cell line KYSE 410-4 by repetitive multicolor fluorescence in situ hybridization

Chromosome aberrations are distinctive features of human malignant tumors. Analysis of chromosomal changes can illuminate the molecular mechanisms underlying the development and progression of cancer. To establish the technique of multicolor fluorescence in situ hybridization （M-FISH） for identifying chromosome aberrations in esophageal carcinoma cell line KYSE 410-4, four pools of 6-color whole-chromosome painting probes have been designed and hybridized on the same metaphase spread by four rounds of repetitive FISH. Repetitive 6-color M-FISH was successfully established and the cytogenetic abnormalities in KYSE 410-4 cells were characterized. Chromosome gains occurred at 2q, 3, 8, 17p, and X. An isochromosome 3q was visualized in the cell line, which might be one intermediate mechanism leading to 3p losses and/or 3q gains. Furthermore, 16 structural arrangements were detected, including four derivative chromosomes. The rearrangement of the centromeric regions accounted for approximately 44% of all rearrangements. The results added a more complete and accurate information of the genetic alterations to the classical cytogenetic description of KYSE 410-4 and provided a detailed cytogenetic background data for appropriate use of the cell line. The established 6-color M-FISH was useful for analyzing chromosomes in the whole genome of human tumors.

Fluorescent aptasensor for detection of live foodborne pathogens based on multicolor perovskite-quantum-dot-encoded DNA probes and dual-stirring-bar-assisted signal amplification

In this study,a fluorescent(FL)aptasensor was developed for on-site detection of live Salmonella typhimurium(S.T.)and Vibrio parahaemolyticus(V.P.).Complementary DNA(cDNA)of aptamer(Apt)-functionalized multicolor polyhedral oligomeric silsesquioxane-perovskite quantum dots(cDNA-POSSPQDs)were used as encoded probes and combined with dual-stirring-bar-assisted signal amplification for pathogen quantification.In this system,bar 1 was labeled with the S.T.and V.P.Apts,and then bar 2 was functionalized with cDNA-POSS-PQDs.When S.T.and V.P.were introduced,pathogen-Apt complexes would form and be released into the supernatant from bar 1.Under agitation,the two complexes reached bar 2 and subsequently reacted with cDNA-POSS-PQDs,which were immobilized on MXene.Then,the encoded probes would be detached from bar 2 to generate FL signals in the supernatant.Notably,the pathogens can resume their free state and initiate next cycle.They swim between the two bars,and the FL signals can be gradually enhanced to maximum after several cycles.The FL signals from released encoded probes can be used to detect the analytes.In particular,live pathogens can be distinguished from dead ones by using an assay.The detection limits and linear range for S.T.and V.P.were 30 and 10 CFU/mL and 10^(2) -10^(6) CFU/mL,respectively.Therefore,this assay has broad application potential for simultaneous on-site detection of various live pathogenic bacteria in water.

Multimodal imaging of experimental choroidal neovascularization

AIM:To compare choroidal neovascularization(CNV)lesion measurements obtained by in vivo imaging modalities,with whole mount histological preparations stained with isolectin GS-IB4,using a murine laser-induced CNV model.METHODS:B6 N.Cg-Tg(Csf1 r-EGFP)1 Hume/J heterozygous adult mice were subjected to laser-induced CNV and were monitored by fluorescein angiography(FA),multicolor(MC)fundus imaging and optical coherence tomography angiography(OCTA)at day 14 after CNV induction.Choroidalretinal pigment epithelium(RPE)whole mounts were prepared at the end of the experiment and were stained with isolectin GS-IB4.CNV areas were measured in all different imaging modalities at day 14 after CNV from three independent raters and were compared to choroidal-RPE whole mounts.Intraclass correlation coefficient(ICC)type 2(2-way random model)and its 95%confidence intervals(CI)were calculated to measure the correlation between different raters’measurements.Spearman’s rank correlation coefficient(Spearman’s r)was calculated for the comparison between FA,MC and OCTA data and histology data.RESULTS:FA(early and late)and MC correlates well with the CNV measurements ex vivo with FA having slightly better correlation than MC(FA early Spearman’s r=0.7642,FA late Spearman’s r=0.7097,and MC Spearman’s r=0.7418),while the interobser ver reliability was good for both techniques(FA early ICC=0.976,FA late ICC=0.964,and MC ICC=0.846).In contrast,OCTA showed a poor correlation with ex vivo measurements(Spearman’s r=0.05716)and high variability between different raters(ICC=0.603).CONCLUSION:This study suggests that FA and MC imaging could be used for the evaluation of CNV areas in vivo while caution must be taken and comparison studies should be performed when OCTA is employed as a CNV monitoring tool in small rodents.

Water-sensitive multicolor luminescence in lanthanide-organic framework for anti-counterfeiting

The developme nt of high-level an ti-co un terfeiti ng tech niq ues is of great sig nifica nee in econo mics and security issues.However,i ntricate read ing methods are required to obta in multi-level info rmati on stored in differe nt colors,which greatly limits the applicati on of an ti-co un terfeit ing tech no logy on sol ving real world problems.Here in,we realize multicolor information anti-counterfeiting under simply external stimulation by utilizing the functional groups and multiple emission centers of lanthanide metal organic framework(Ln-MOFs)to tune luminescence color.Water responsive multicolor luminescence represented by both the tunable color from red to blue within the visible region and high sensitive responsivity has bee n achieved,owing to the in creased nonr adiative decay pathways and enhan ced Eu3+-to-liga nd en ergy back tra nsfer.Remarkably,i nfo rmatio n hidde n in differe nt colors n eeds to be read with a specific water content,which can be used as an en crypti on key to en sure the security of the info rmati on for high-level an ti-co un terfeiti ng.

Experimental Research on the SizeMeasurement of the High Temperature ForgingBased on Multicolor CCD Technology

In order to determine the size measurement accuracy of the high temperature forging＇s multicolor CCD image by using computerprograms, this paper obtained the high temperature forging＇s CCD image by multicolor CCD camera and its fact size by thevernier caliper on the forging field, and then measured the size of the high temperature forging from its CCD image, compared thesize from the CCD image and the size from the vernier caliper, the result shows that the measurement accuracy satisfied theindustrial production.

A novel cytogenetic abnormality r(7)(::p11.2->q36.3::) in a Philadelphia-positive chronic myeloid leukemia case

The so-calledPhiladelphia(Ph) chromosome is present in more than 90% of chronic myeloid leukemia (CML) cases. It results in juxtaposition of the 5' part of the BCR gene on chromosome 22 and the 3' part of the ABL1 gene on chromosome 9. An additional acquired monosomy 7 or deletion of 7q is associated with poor prognosis in a variety of myeloid disorders. Here we report a novel Ph chromosome positive CML case with a ring chromosome 7 [r(7)]. Immunophenotyping was compatible with CML, although 4.5% of total leucocytes appeared like acute myelogeneous leukemia (AML) subtype M2. The r(7) was characterized in detail by array-proven multicolor banding (aMCB), the latter being of enormous significance to characterize breakpoint regions in detail. Underlying mechanisms and prognostic are discussed, as ring chromosomes are rare cytogenetic abnormalities in hematopoietic malignancies.

Multicolor Circularly Polarized Luminescence of a Single-Component System Revealing Multiple Information Encryption

Metal-free materials with multicolor tunable circularly polarized luminescence(CPL)are attractive because of their potential applications in information storage and encryption.Here,we designed two enantiomers composed of chiral dialkyl glutamides and achiral vibration-induced emission(VIE)moiety,which can switch on CPL after a simple gelation process.It is noteworthy that the CPL colors vary in different solvents,and this is attributed to various self-assembly-induced microstructures,in which the VIE moiety is restrained to different degrees.Accordingly,a multidimensional code system composed of a quick response code,a ultraviolet(UV)light-activated color code,and a CPL information figure was constructed.To our satisfaction,the system possesses multiple information-storage functions.The orthogonal anticounterfeiting and CPLenhanced encryption functions also improve the system information encryption ability.In brief,this study provides a practical example of CPL applied to information security and an effective approach to obtain a single-component color-tunable CPL material with multiple information storage and encryption functions as well.

Synergistic fluorescent hydrogel actuators with selective spatial shape/color-changing behaviors via interfacial supramolecular assembly

Biomimetic intelligent polymeric hydrogel actuators with cooperative fluorescence-color switchable behaviors are expected to find great potential applications in soft robotics,visual detection/display,and camouflage applications.However,it remains challenging to realize the spatial manipulation of synergistic shape/color-changing behaviors.Herein,we report an interfacial supramolecular assembly(ISA)approach that enables the construction of robust fluorescent polymeric hydrogel actuators with spatially anisotropic structures.On the basis of this ISA approach,diverse 2D/3D soft fluorescent hydrogel actuators,including chameleon-and octopi-shaped ones with spatially anisotropic structures,were facilely assembled from two different fluorescent hydrogel building blocks sharing the same physically cross-linked agar network.Spatially control over synergistic shape/color-changing behaviors was then realized in one single anisotropic hydrogel actuator.The proposed ISA approach is universal and expected to open promising avenues for developing powerful bioinspired intelligent soft actuators/robotics with selective spatial shape/color-changing behaviors.

Dihydroanthracene bridged bis-naphthopyrans:A multimodal chromophore with mechano-and photo-chromic properties

Mechanochromophores based on bichromic molecular switches,such as bis-naphthopyanes,allow multimodal mechanochromic behavior beyond the typical binary response from single chromophores,which is important for distinguishing between multiple stress states through discrete changes in color.Spontaneously generated persistent and distinguishable multi-colors from activated bis-naphthopyanes remain challenging.And the versatility of bis-mechanophore design for advanced optical molecular systems and the fundamental insights into the corresponding mechano-reactivity are not enough.Here,we identify a dihydroanthracene bridged bis-naphthopyrans as a multimodal mechanochromophore in polymers.Bridging two pyrans with the sterically constrained dihydroanthracene is helpful to control the steric effect for the favorable formation of a distinctly appreciable bis-merocyanine(bis-MC)product.By varying the length of the polymer chains,the force delivered to the mechanophore is modulated,resulting in a gradient change in the relative distribution of two distinctly colored MC products and a multicolor mechanochromism.Mechanical activation of this bis-naphthopyanes proceeds via a mechanistically distinct pathway compared to the photochemical process.In addition,the bulk films can also achieve pronounced color changes when subjected to mechanical force.This study thus further expands the molecular diversity of mechanochromophores and tune the multimodal switch properties of bis-naphthopyrans based polymers.

Cool white light and tunable multicolor emission from Tb^(3+)/Dy^(3+)co-activated glasses under different excitations for WLEDs

A set of co-doped(Tb^(3+)/Dy^(3+))lithium zinc borate(LZB)glasses were developed by melt quenching.The structural evaluation was performed for synthesized glassy matrices.The Dy^(3+)and Tb^(3+)individually doped glasses exhibit intense yellow and green luminescence bands at 575 nm(^(4)F_(9/2)→^(6)H_(13/2))and543 nm(^(5)D_(4)→^(7)F_(5)),respectively.The sensitization effect of Dy^(3+)on Tb^(3+)was analyzed by increasing the Tb^(3+)content with respect to the optimum Dy^(3+)content(0.5 mol%)in Dy^(3+)/Tb^(3+).The spectral overlay of Dy^(3+)luminescence and Tb^(3+)absorption profiles,Dy^(3+)/Tb^(3+)PL spectra under different excitations 352,362,376,and 385 nm,shortening decay lifetimes of Dy^(3+)in Dy^(3+)/Tb^(3+)co-activated glasses,energy transfer(ET)parameters,chromaticity coordinates and their corresponding correlated temperatures all help to explain ET from Dy^(3+)to Tb^(3+).At 385 nm of Dy^(3+)excitation,the optimized co-activated(0.5Dy^(3+)+1.0Tb^(3+)):LZB glass displays cool white light emission.The non-radiative ET from Dy^(3+)to Tb^(3+)is dominated by electric dipole-dipole interaction and its ET efficiency was calculated to be 63%.At the same time,reverse ET from Tb^(3+)to Dy^(3+)was also analyzed.The shift in color coordinates from dominant yellow to greenish-yellow,green and white light emission suggests that Dy^(3+)/Tb^(3+)coactivated LZB glasses can be a potential candidate for UV converted multicolor and white light emitting devices.

Multicolor upconversion reversible modulations in YNbO_(4):Er^(3+)/Tm^(3+)/Yb^(3+) photochromic materials

Photochromic materials with multicolor upconversion reversible modulations are attractive in optical switching devices.Herein,the fabricated YNbO_(4):Er^(3+)/Tm^(3+)/Yb^(3+) materials exhibit excellent photochromism and multicolor upconversion properties from green,red to near infrared(NIR) emissions with increasing Yb concentrations.Reversible multiband upconversion modulations are achieved by alternating light(365 and 405 nm) or thermal stimuli.After 365 nm irradiation,the luminescence color changes from yellow to red,the luminescent photoswitching contrast reaches up to 86.21%(green),82.12%(red) and 77.38%(NIR) in the Y_(0.83)Er_(0.01)Tm_(0.01)NbO_(4):0.15 Yb sample.Besides,the upconversion emission intensity before and after photochromic reaction shows remarkable change in a wide temperature range of 298-718 K.These results indicate that the Er^(3+)/Tm^(3+)/Yb^(3+) tri-doped YNbO_(4) materials can be a good candidate in optical switching and data storage applications.

Complementary multicolor electrochromic devices with excellent stability based on porous tin oxide nanosheet scaffold

Electrochromic devices(ECDs)have been extensively investigated as promising candidates in broad cutting-edge applications,such as smart windows,electronic labels,adaptive camouflage,etc.However,they have suffered from either inadequate color variations or poor cycling stability for a long time.Herein,we developed a general strategy to boost the cyclic stability and enrich the color variations of ECDs by scrupulous design of the composition and nanostructure of electrodes,in which porous tin oxide(SnO_(2))nanosheets serve as the scaffold and typical metal oxides or conducting polymers as the active electrochromic materials.Various electrochromic composite materials,including polyaniline(PANI)@SnO_(2),V2O5@SnO_(2),and WO_(3)@SnO_(2) heterostructured nanoarrays were prepared by the facile wet-chemical method.These composite electrodes exhibit remarkable electrochromic performances,e.g.,superior cycling stability(more than 2000 cycles),rich color variations(more than 5 colors for PANI@SnO_(2)),and enlarged optical modulation.These excellent performances account for the heterogenous porous nanoarrays,which not only facilitate the intercalation/extraction of ions but also relieve the stress generated during the electrochemical process.In addition,diverse prototypes of complementary multicolor ECD with excellent cycling stability(over thousands of cycles)and rich color variations(8 colors)were realized for the first time.We believe that our work put forward a general strategy for developing high-quality multicolor complementary electrochromic devices.

Triggering triplet excitons of carbon nanodots through nanospace domain confinement for multicolor phosphorescence in aqueous solution

Easy non-radiative decay property of long-lived triplet excitons in aqueous solution obstructs their applications in aquatic surroundings.Recently reported phosphorescence phenomena in aqueous solution have excited researchers enormously but achieving full-color water-soluble phosphorescent carbon nanodots(CNDs)is still a challenging issue.Herein,full-color phosphorescence of water-soluble CNDs has been demonstrated by triggering their triplet excitons through nanospace domain confinement,and Förster energy resonance transfer is used for further tuning phosphorescence range.The phosphorescence spans across most of the visible spectrum,ranging from 400 to 700 nm.In an aqueous solution,the CNDs exhibits blue,green,and red phosphorescence,lasting for approximately 6,10,and 7 s,respectively.Correspondingly,the phosphorescence quantum yields are 11.85%,8.6%and 3.56%,making them readily discernible to the naked eyes and laying a solid foundation for practical application.Furthermore,phosphorescence flexible optical display and bioimaging have been demonstrated by using the multicolor CNDs-based nanomaterials,showing distinct superiority for accuracy and complete display and imaging in complex emission background.

Controlling molecular assembly on time scale:Time-dependent multicolor fluorescence for information encryption

Dynamic assembly on time scale is common in biological systems but rare for artificial materials,especially for smart luminescent materials.Programming molecular assembly in a spatio-temporal manner and resulting in white-light-including multicolor fluorescence with time-dynamic features remains challenging.Herein,controlling molecular assembly on time scale is achieved by integrating a pH-responsive motif to a transient alkaline solution which is fabricated by activators(NaOH)and deactivators(esters),leading to automatic assembly on time scale and time-dependent multicolor fluorescence changing from blue to white and yellow.The kinetics of the assembly process is dependent on the ester hydrolysis process,which can be controlled by varying ester concentrations,temperature,initial pH,stirring rate and ester structures.This dynamic fluorescent system can be further developed for intelligent fluorescent materials such as fluorescent ink,three-dimension(3D)codes and even four-dimension(4D)codes,exhibiting a promising potential for information encryption.

Dynamic Crosslinked Phosphorescent Poly(vinyl alcohol)-Terpyridine Films with Enhanced Mechanical Properties and Tunable Shape Memory

Achieving versatile room temperature phosphorescence(RTP)materials,especially with tunable mechanical properties and shape memory is attractive and essential but rarely reported.Here,a strategy was reported to realize multi-functional RTP films with multicolor fluorescence,ultralong afterglow,adjustable mechanical properties,and shape memory through the synergistic dynamic interaction of lanthanide(Ln~Ⅲ)-terpyridine coordination,borate ester bonds,and hydrogen bondings in a poly(vinyl alcohol)(PVA)matrix.By varying the amount of borax,the mechanical properties of the films could be finely controlled due to the change of crosslinking degree of dynamic borate ester bonds in PVA.The assembly and disassembly of borate ester bonds upon the trigger of borax and acid were applied as reversible linkage to achieve programmable shape memory behavior.In addition,the films displayed both fascinating multicolor fluorescence and ultralong afterglow characteristics due to the presence of Ln III doping and confinement of terpyridine in PVA.This study provides a new avenue to impart modulable mechanical strength and shape memory to RTP materials.

AIE gel exhibiting continuous gradient fluorescence based on a polar-responsive AIE-gen

Traditional multicolor fluorescent hydrogels are generated through the assembly of discrete fluorescent hydrogels,which is not a complete integration much distinct from living organisms.On the basis of aggregation-induced emission(AIE),a special solvent polar-responsive AIE molecule possessing a twisted intramolecular charge transfer(TICT)effect was noticed.By incorporating it into the gel network,an AIE gel that displays continuous gradient fluorescence was fabricated.First,hydrogel A containing the solvent polar-responsive AIE-gen was prepared to show orange fluorescence.After soaking in the organic solvents,the fluorescence color transition of hydrogel A ranging from orange to green occurred when being immersed in high-polarity organic solvents ascribed to the embedded AIE-gen owning TICT effect.Then,hydrogel A was successively lifted up from organic solvents.Due to the different immersion time of each section for the hydrogel,the polarity difference occurred.Then,the produced gel B showed continuous gradient fluorescence ranging from orange to green under the irradiation of UV light.

Multicolor V_(2)O_(5)/TiO_(2)electrochromic films with fast switching and long lifespan for camouflage and information display

V_(2)O_(5),which has multicolor and energy storage properties,is a promising electrochromic material for multifunctional electrochromic devices,but its practical application is limited by its poor lifespan and long switching time.In this work,high-performance V_(2)O_(5)/TiO_(2)films were fabricated by spraying a V_(2)O_(5)solution on in situ-grown TiO_(2)nanorods.Due to the porous structure formed between the TiO_(2)nanorods and the remarkable electron transfer performance of TiO_(2),the switching time of the V_(2)O_(5)/TiO_(2)films decreased.Moreover,the strong adhesion between the TiO_(2)nanorods and F-doped tin oxide(FTO)glass and the increased surface roughness of the substrates significantly improved the cycling stability of the V_(2)O_(5)/TiO_(2)films.With a large transmittance modulation(47.8%at 668 nm),fast response speed(τ_(c)=5.1 s,τ_(b)=4.2 s),and long lifespan,V_(2)O_(5)/TiO_(2)films were used as electrodes for the electrochromic energy storage device(EESD),which switched in six colors through color overlay:dark orange,sandy yellow,green-yellow,yellow-green,dark green,and dark brown.Inspired by pixel displays,EESDs were designed by segmenting V_(2)O_(5)films to stagger the display of the electrochromic and ion storage layers,which presented 11 types of information based on different combinations of colors.This work provides inspiration for developing multifunctional electrochromic devices,especially for camouflage and information displays.