A compact volume holographic imaging(VHI)method that can detect fluorescence objects located in diffusive medium in spectral selective imaging manner is presented.The enlargement of lateralfield of view of the VHI sys...A compact volume holographic imaging(VHI)method that can detect fluorescence objects located in diffusive medium in spectral selective imaging manner is presented.The enlargement of lateralfield of view of the VHI system is realized by using broadband illumination and demagnification optics.Each target spectrum of°uorescence emitting from a di®usive medium is probed by tuning the inclination angle of the transmission volume holographic grating(VHG).With the use of the single transmission VHG,fluorescence images with different spectrum are obtained sequentially and precise three-dimensional(3D)information of deep fluorescent objects located in a diffusive medium can be reconstructed from these images.The results of phantom experiments demonstrate that two fluorescent objects with a sub-millimeter distance can be resolved by spectral selective imaging.展开更多
Fluorescence molecular imaging enables the visualization of basic molecular processes such as gene expression,enzyme activity,and disease-specific molecular interactions in vivo using targeted contrast agents,and ther...Fluorescence molecular imaging enables the visualization of basic molecular processes such as gene expression,enzyme activity,and disease-specific molecular interactions in vivo using targeted contrast agents,and therefore,is being developed for early detection and in situ characterization of breast cancers.Recent advances in developing near-infrared fluorescent imaging contrast agents have enabled the specific labeling of human breast cancer cells in mouse model systems.In synergy with contrast agent development,this paper describes a needle-based fluorescence molecular imaging device that has the strong potential to be translated into clinical breast biopsy procedures.This microendoscopy probe is based on a gradient-index(GRIN)lens interfaced with a laser scanning microscope.Specifications of the imaging performance,including the field-of-view,transverse resolution,and focus tracking characteristics were calibrated.Orthotopic MDA-MB-231 breast cancer xenografts stably expressing the tdTomato red fluorescent protein(RFP)were used to detect the tumor cells in this tumor model as a proof of principle study.With further development,this technology,in conjunction with the development of clinically applicable,injectable fluorescent molecular imaging agents,promises to perform fluorescence molecular imaging of breast cancers in vivo for breast biopsy guidance.展开更多
Near-infraredfluorescence(NIRF)imaging involves the separation of weakfluorescence signals from backscattered excitation light.The measurement sensitivity of current NIRF imaging systems is limited by the excitation l...Near-infraredfluorescence(NIRF)imaging involves the separation of weakfluorescence signals from backscattered excitation light.The measurement sensitivity of current NIRF imaging systems is limited by the excitation light leakage through rejectionfilters.In this contribution,the authors demonstrate that the excitation light leakage can be suppressed upon using appropriatefilter combination and permutations.The excitation light leakage and measurement sensitivity were assessed and compared in this study by computing the transmission ratios of excitation to emission light collected and the signal-to-noise ratios in well-controlled phantom studies with differentfilter combinations and permutations.Using appropriatefilter combinations and permutations,we observe as much as two orders of magnitude reduction in the transmission ratio and higher signal-to-noise ratio.展开更多
Bioluminescence tomography(BLT)is a novel opt ical molecular imaging technique that advanced the conventional planar bioluminescence imaging(BLI)into a quantifiable three-dimensional(3D)approach in preclinical living ...Bioluminescence tomography(BLT)is a novel opt ical molecular imaging technique that advanced the conventional planar bioluminescence imaging(BLI)into a quantifiable three-dimensional(3D)approach in preclinical living animal studies in oncology.In order to solve the inverse problem and reconstruct tumor lesions inside animal body accurately,the prior structural information is com-monly obtained from X ray computed tomography(CT).This strategy requires a complicated hybrid imaging system,extensive post imaging analysis and involvement of ionizing radiation.Moreover,the overall robustness highly depends on the fusion accuracy between the optical and structural information.Here,we present a pure optical bioluminescence tomographic(POBT)system and a novel BLT workfow based on multi-view projection acquisition and 3D surface reconstruction.This met hod can reconstruct the 3D surface of an imaging subject based on a sparse set of planar white-light and bioluminescent images,so that the prior structural information can be offered for 3D tumor lesion reconstruction without the involvement of CT.The performance of this novel technique was evaluated through the comparison with a conventional dual-modality tomo-graphic(DMT)system and a commercialized optical imaging system(IVIS Spectrum)using three breast cancer xenografts.The results revealed that the new technique offered comparable in vivo tomographic accuracy with the DMT system(P>0.05)in much shorter data analysis time.It also offered significantly better accuracy comparing with the IVIS system(P<0.04)without sacrificing too much time.展开更多
For fluorescence molecular tomography(FMT),image quality could be improved by incorporating a sparsity constraint.The L1 norm regularization method has been proven better than the L2 norm,like Tikhonov regularization....For fluorescence molecular tomography(FMT),image quality could be improved by incorporating a sparsity constraint.The L1 norm regularization method has been proven better than the L2 norm,like Tikhonov regularization.However,the Tikhonov method was found capable of achieving a similar quality at a high iteration cost by adopting a zeroing strategy.By studying the reason,a Tikhonov-regularization-based projecting sparsity pursuit method was proposed that reduces the iterations significantly and achieves good image quality.It was proved in phantom experiments through time-domain FMT that the method could obtain higher accuracy and less oversparsity and is more applicable for heterogeneous-target reconstruction,compared with several regularization methods implemented in this Letter.展开更多
There is a growing realization that cell-to-cell variations in gene expression have importantbiological consequences underlying phenotype diversity and cell fate. Although analytical toolsfor measuring gene expression...There is a growing realization that cell-to-cell variations in gene expression have importantbiological consequences underlying phenotype diversity and cell fate. Although analytical toolsfor measuring gene expression, such as DNA microarrays, reverse-transcriptase PCR and in situhybridization have been widely utilized to discover the role of genetic variations in governingcellular behavior, these methods are performed in cell lysates and/or on fixed cells, and thereforelack the ability to provide comprehensive spatial-dynamic information on gene expression. Thishas invoked the recent development of molecular imaging strategies capable of illuminatingthe distribution and dynamics of RNA molecules in living cells. In this review, we describe aclass of molecular imaging probes known as molecular beacons (MBs), which have increasinglybecome the probe of choice for imaging RNA in living cells. In addition, we present the majorchallenges that can limit the ability of MBs to provide accurate measurements of RNA, anddiscuss efforts that have been made to overcome these challenges. It is envisioned that withcontinued refinement of the MB design, MBs will eventually become an indispensable tool foranalyzing gene expression in biology and medicine.展开更多
Fluorescence imaging is a noninvasive and dynamic real-time imaging technique;however,it exhibits poor spatial resolution in centimeter-deep tissues because biological tissues are highly scattering media for optical r...Fluorescence imaging is a noninvasive and dynamic real-time imaging technique;however,it exhibits poor spatial resolution in centimeter-deep tissues because biological tissues are highly scattering media for optical radiation.The recently developed ultrasound-controlled fluorescence(UCF)imaging is a novel imaging technique that can overcome this bottleneck.Previous studies suggest that the effective contrast agent and sensitive imaging system are the two pivotal factors for generating high-resolution UCF images ex vivo and/or in vivo.Here,this review highlights the recent advances(2015e2020)in the design and synthesis of contrast agents and the improvement of imaging systems to realize high-resolution UCF imaging of deep tissues.The imaging performances of various UCF systems,including the signal-to-noise ratio,imaging resolution,and imaging depth,are specifically discussed.In addition,the challenges and prospects are highlighted.With continuously increasing research interest in this field and emerging multidisciplinary applications,UCF imaging with higher spatial resolution and larger imaging depth may be developed shortly,which is expected to have a far-reaching impact on disease surveillance and/or therapy.展开更多
In this study we implemented an axillary SLN invasion model to develop highly sensitive imaging strategies enabling detection of a very small amount of tumor cells. A highly diffusible molecular probe targeting &a...In this study we implemented an axillary SLN invasion model to develop highly sensitive imaging strategies enabling detection of a very small amount of tumor cells. A highly diffusible molecular probe targeting αvβ3 and αvβ5 integrins was investigated either via IV or locoregional injections. We additionally documented the potential interferences of this Near Infrared Fluorescence Probe with Blue Patente V and ICG dyes routinely used to facilitate lymph node detection during surgery. The human mammary adenocarcinoma MDA-MB-231-luc model was injected into the forepaw of nude female rats to obtain a controlled invasion of the axillary LN. Thanks to its high sensitivity, BLI was selected to achieve in vivo quantitation of tumor cells in SLNs and determine eligible animals for the study. NIRF of integrins was performed at 680 nm both in vivo and ex vivo using spectral unmixing to suppress auto-fluorescence signal and preserve sensitivity. In vivo BLI was quite reliable in estimating discrete invasion by cancer cells in the LN with thresholds of detection and quantitation of about 500 and 1500 cells respectively. For fluorescence at 680 nm, in vivo imaging is not suitable to detect micro-invasion, but ex vivo fluorescence with spectral unmixing of SLNs confirmed the presence of a tumor burden as low as 1500 cells expressing αvβ3/αvβ5 integrins. Targeting few tumor cells inside a micro-invaded sentinel lymph node by molecular probes is not sensitive enough to provide direct in vivo or peroperative imaging. At the time NIRF is performed on the excised specimen, high sensitivity imaging associated with spectral unmixing allowed such detection within less than 1 minute of examination.展开更多
We report on tests of combined positron emission tomography(PET)andfluorescence molecular tomography(FMT)imaging system for in vivo investigation on small animals.A nude mouse was inoculated with MD-MB-231 breast canc...We report on tests of combined positron emission tomography(PET)andfluorescence molecular tomography(FMT)imaging system for in vivo investigation on small animals.A nude mouse was inoculated with MD-MB-231 breast cancer cells which expressed redfluorescent protein(RFP).For FMT system,reflective illumination mode was adopted with full-angle data acquisition.[18F]-Fluorodeoxyglucose([18F]-FDG)was used as radioactive tracer for PET.Both data were acquired simultaneously and then reconstructed separately before fusion.Fluorescent tomography results showed exactly where the tumor was located while PET results offered more metabolic information.Results confirmed feasibility for tumor detection and showed superiority to single modality imaging.展开更多
Molecular imaging(MI)is a novel imaging discipline that has been continuously developed in recent years.It combines biochemistry,multimodal imaging,biomathematics,bioinformatics,cell&molecular physiology,biophysic...Molecular imaging(MI)is a novel imaging discipline that has been continuously developed in recent years.It combines biochemistry,multimodal imaging,biomathematics,bioinformatics,cell&molecular physiology,biophysics,and pharmacology,and it provides a new technology platform for the early diagnosis and quantitative analysis of diseases,treatment monitoring and evaluation,and the development of comprehensive physiology.Fluorescence Molecular Tomography(FMT)is a type of optical imaging modality in MI that captures the three-dimensional distribution of fluorescence within a biological tissue generated by a specific molecule of fluorescent material within a biological tissue.Compared with other optical molecular imaging methods,FMT has the characteristics of high sensitivity,low cost,and safety and reliability.It has become the research frontier and research hotspot of optical molecular imaging technology.This paper took an overview of the recent methodology advances in FMT,mainly focused on the photon propagation model of FMT based on the radiative transfer equation(RTE),and the reconstruction problem solution consist of forward problem and inverse problem.We introduce the detailed technologies utilized in reconstruction of FMT.Finally,the challenges in FMT were discussed.This survey aims at summarizing current research hotspots in methodology of FMT,fromwhich future research may benefit.展开更多
On May 6, 2009, the X-ray imaging and biomedical application beamline(BL13W1) at Shanghai Synchrotron Radiation Facility(SSRF) officially opened to users, with 8–72.5 ke V X-rays. The experimental station is equipped...On May 6, 2009, the X-ray imaging and biomedical application beamline(BL13W1) at Shanghai Synchrotron Radiation Facility(SSRF) officially opened to users, with 8–72.5 ke V X-rays. The experimental station is equipped with four sets of X-ray CCD detectors of different pixel size(0.19–24 μm) for on-line phase-contrast imaging and micro-CT imaging with 0.8 μm spatial resolution and 1 ms temporal resolution. An in vivo microCT experiment for a living insect was realized in 4 s. An X-ray fluorescence detector is equipped for X-ray fluorescence mapping imaging and X-ray fluorescence micro-CT imaging with 50 μm spatial resolution. In order to meet different requirements from the users, several experimental methods, such as X-ray spiral micro-CT, Xray local micro-CT, X-ray fast micro-CT, X-ray grating-based differential micro-CT, X-ray fluorescence microCT and X-ray quantitative micro-CT have been developed, and nearly 60 papers related to those developments for this beamline have been published. Moreover, the beamline has realized the remote fast CT reconstruction,providing a great convenience for the users to process experimental data at their offices. As of August 2014,the beamline has offered the user beamtime of(23 145 h), from which 232 user papers have been published,including 151 SCI papers and 55 papers with SCI impact factor > 3. The quantity and quality of the user paper outcome keep a steady increase. Some typical user experimental results are introduced.展开更多
Molecular imaging has emerged as a new discipline in gastrointestinal endoscopy.This technology encompasses modalities that can visualize disease-specific morphological or functional tissue changes based on the molecu...Molecular imaging has emerged as a new discipline in gastrointestinal endoscopy.This technology encompasses modalities that can visualize disease-specific morphological or functional tissue changes based on the molecular signature of individual cells.Molecular imaging has several advantages including minimal damage to tissues,repetitive visualization,and utility for conducting quantitative analyses.Advancements in basic science coupled with endoscopy have made early detection of gastrointestinal cancer possible.Molecular imaging during gastrointestinal endoscopy requires thedevelopment of safe biomarkers and exogenous probes to detect molecular changes in cells with high specificity anda high signal-to-background ratio.Additionally,a high-resolution endoscope with an accurate wide-field viewing capability must be developed.Targeted endoscopic imaging is expected to improve early diagnosis and individual therapy of gastrointestinal cancer.展开更多
Photosensitive fluorescent probes have become powerful tools in chemical biology and molecular biophysics,which are used to investigate cellular processes with high temporal and spatial resolution.Accordingly,photosen...Photosensitive fluorescent probes have become powerful tools in chemical biology and molecular biophysics,which are used to investigate cellular processes with high temporal and spatial resolution.Accordingly,photosensitive fluorescent probes,including photoactivatable,photoconvertible,and photoswitchable fluorophores,have been extensively developed during the past decade.The photoswitchable fluorophores have received much attention because they highlight cellular events clearly.This minireview summarizes recent advances of using reversibly photoswitchable fluorophores and their applications in innovative bioimaging.Photoswitchable fluorophores include photoswitchable fluorescent proteins,photoswitchable fluorescent organic molecules(dyes),and photoswitchable fluorescent nanoparticles.Several strategies have been developed to synthesize photoswitchable fluorophores,including engineering combination proteins,chemical synthesis,polymerization,and self-assembly.Here we concentrate on polymer nanoparticles with optically switchable emission properties:either fluorescence on/offor dualalternating-color fluorescence photoswitching.The essential mechanisms of fluorescence photoswitching enable different types of photoswitchable fluorophores to change emission intensity or wavelength(color)and thus validating the basis of the fluorescence on/offor dual-color photoswitching design.Generally the possible applications of any fluorophores are to label biological targets,followed by specific imaging.The newly developed photoswitchable fluorophores enable super-resolution fluorescence imaging because of their photosensitive emission.Finally,we summarize the important area regarding future research and development on photoswitchable fluorescent nanoparticles.展开更多
Endogenous biomolecules in cells are the basis of all life activities.Directly visualizing the structural characteristics and dynamic behaviors of cellular biomolecules is signiffcant for understanding the molecular m...Endogenous biomolecules in cells are the basis of all life activities.Directly visualizing the structural characteristics and dynamic behaviors of cellular biomolecules is signiffcant for understanding the molecular mechanisms in various biological processes.Singlemolecule localization microscopy(SMLM)can circumvent the optical diffraction limit,achieving analysis of the ffne structures and biological processes in living cells with nanoscale resolution.However,the large size of traditional imaging probes prevents SMLM from accurately locating ffne structures and densely distributed biomolecules within cells.In recent years,nucleic acid probes have emerged as potential tools to replace conventional SMLM probes by virtue of their small size and high speciffcity.In addition,due to their programmability,nucleic acid probes with different conformations can be constructed via sequence design,further extending the application of SMLM in bioanalysis.Here,we discuss the design concepts of different conformational nucleic acid probes for SMLM and summarize the application of SMLM based on nucleic acid probes in the ffeld of biomolecules.Furthermore,we provide a summary and future perspectives of the nucleic acid probe-based SMLM technology,aiming to provide guidance for the acquisition of nanoscale information about cellular biological processes.展开更多
A two-stage source reconstruction algorithm for bioluminescence tomography (BLT) is developed using hybrid finite element method (FEM). The proposed algorithm takes full advantages of linear and quadratic FEMs, which ...A two-stage source reconstruction algorithm for bioluminescence tomography (BLT) is developed using hybrid finite element method (FEM). The proposed algorithm takes full advantages of linear and quadratic FEMs, which can be used to localize and quantify bioluminescent source accurately. In the first stage, a large permissible region is roughly determined and then iteratively evolved to reduce matrix dimension using efficient linear FEM. In the final stage, high-convergence quadratic FEM is applied to improve reconstruction result. Both numerical simulation and physical experiment are performed to evaluate the proposed algorithm. The relevant results demonstrate that quantitative reconstruction can be well achieved in terms of computation efficiency, source position, power density, and total power when compared with previous studies.展开更多
Laminar optical tomography(LOT)is a mesoscopic tomographic imaging technique ranging between confocal microscopy and diffuse optical tomography(DOT).Fluorescence LOT(FLOT)provides depth-resolved molecular information ...Laminar optical tomography(LOT)is a mesoscopic tomographic imaging technique ranging between confocal microscopy and diffuse optical tomography(DOT).Fluorescence LOT(FLOT)provides depth-resolved molecular information with 100-200μm resolution over 2-3mm depth.In this study,we use Monte Carlo simulation and singular-value analysis(SVA)to optimize the source-detector configurations for potential enhancement of FLOT imaging performance.The effects of different design parameters,including source incidence and detector collection angles,detector number,and sampling density,are presented.The results indicate that angled incidence/detection configuration might improve the imaging resolution and depth sensitivity,especially for low-scattering medium.Increasing the number of detectors and the number of scanning steps will also result in enhanced imaging performance.We also demonstrate that the optimal imaging performance depends upon the background scattering coe±cient.Our result might provide an optimization strategy for FLOT or LOT experimental setup.展开更多
The real-time, noninvasive, nonionizing, high spatiotemporal resolution, and flexibility characteristics of molecular fluorescence imaging provide a uniquely powerful approach to imaging and monitoring the physiology ...The real-time, noninvasive, nonionizing, high spatiotemporal resolution, and flexibility characteristics of molecular fluorescence imaging provide a uniquely powerful approach to imaging and monitoring the physiology and pathophysiology of ischemic stroke. Currently, various fluorescence probes have been synthesized with the aim of improving quantitative and quantitative studies of the pathologic processes of ischemic stroke in living animals. In this review, we present an overview of current activatable fluorescence probes for the imaging and diagnosis of ischemic stroke in animal models. We categorize the probes based on their activatable signals from the biomarkers associated with ischemic stroke, and we present representative examples of their functional mechanisms. Finally, we briefly discuss future perspectives in this field.展开更多
Fluorescence liftime imaging (FLIM) of modified hydrophobic bodipy dyes that act as fluorescent molecular rotors shows that the fluorescence lifetime of these probes is a function of the microviscosity of their envi...Fluorescence liftime imaging (FLIM) of modified hydrophobic bodipy dyes that act as fluorescent molecular rotors shows that the fluorescence lifetime of these probes is a function of the microviscosity of their environment. Incubating cells with these dyes, we find a punctate and continuous distribution of the dye in cells. The viscosity value obtained in what appears to be endocytotic vesicles in living cells is around 100 times higher than that of water and of cellular cytoplasm.Time-resolved fluorescence anisotropy measurements also yield rotational correlation times consistent with large microviscosity values. In this way, we successfully develop a practical and versatile approach to map the microviscosity in cells based on imaging fluorescent molecular rotors.展开更多
基金This work is supported by the National Basic Research Program of China(973)under Grant No.2011CB707701the National Natural Science Foundation of China under Grant Nos.61361160418,61322101,81227901,81271617,and 61401246the National Major Scientific Instrument and Equipment Development Project under Grant No.2011YQ030114.
文摘A compact volume holographic imaging(VHI)method that can detect fluorescence objects located in diffusive medium in spectral selective imaging manner is presented.The enlargement of lateralfield of view of the VHI system is realized by using broadband illumination and demagnification optics.Each target spectrum of°uorescence emitting from a di®usive medium is probed by tuning the inclination angle of the transmission volume holographic grating(VHG).With the use of the single transmission VHG,fluorescence images with different spectrum are obtained sequentially and precise three-dimensional(3D)information of deep fluorescent objects located in a diffusive medium can be reconstructed from these images.The results of phantom experiments demonstrate that two fluorescent objects with a sub-millimeter distance can be resolved by spectral selective imaging.
基金the Nano-Biotechnology Award of the State of Maryland,the Minta Martin Foundation,the General Research Board(GRB)Award of the University of Maryland,and the University of Maryland Baltimore(UMB)and College Park(UMCP)Seed Grant Program,and the Prevent Cancer Foundation(to Y.C.)Support from NIH P50 CA103175(JHU ICMIC Program,to V.R.)NIH CA134695(to K.G.)is gratefully acknowledged.
文摘Fluorescence molecular imaging enables the visualization of basic molecular processes such as gene expression,enzyme activity,and disease-specific molecular interactions in vivo using targeted contrast agents,and therefore,is being developed for early detection and in situ characterization of breast cancers.Recent advances in developing near-infrared fluorescent imaging contrast agents have enabled the specific labeling of human breast cancer cells in mouse model systems.In synergy with contrast agent development,this paper describes a needle-based fluorescence molecular imaging device that has the strong potential to be translated into clinical breast biopsy procedures.This microendoscopy probe is based on a gradient-index(GRIN)lens interfaced with a laser scanning microscope.Specifications of the imaging performance,including the field-of-view,transverse resolution,and focus tracking characteristics were calibrated.Orthotopic MDA-MB-231 breast cancer xenografts stably expressing the tdTomato red fluorescent protein(RFP)were used to detect the tumor cells in this tumor model as a proof of principle study.With further development,this technology,in conjunction with the development of clinically applicable,injectable fluorescent molecular imaging agents,promises to perform fluorescence molecular imaging of breast cancers in vivo for breast biopsy guidance.
基金supported by the National Institutes of Health,R01 CA112679 and U54 CA136404 and the Texas Star Award.
文摘Near-infraredfluorescence(NIRF)imaging involves the separation of weakfluorescence signals from backscattered excitation light.The measurement sensitivity of current NIRF imaging systems is limited by the excitation light leakage through rejectionfilters.In this contribution,the authors demonstrate that the excitation light leakage can be suppressed upon using appropriatefilter combination and permutations.The excitation light leakage and measurement sensitivity were assessed and compared in this study by computing the transmission ratios of excitation to emission light collected and the signal-to-noise ratios in well-controlled phantom studies with differentfilter combinations and permutations.Using appropriatefilter combinations and permutations,we observe as much as two orders of magnitude reduction in the transmission ratio and higher signal-to-noise ratio.
基金the National Basic Research Program of China(973 Program)under Grant No.2015CB755500the National Natural Science Foundation of China under Grant No.81227901,61231004,81527805 and 61401462+3 种基金the Scienti¯c Research and Equipment Development Project of the Chinese Academy of Sciences under Grant No.YZ201359the Chinese Academy of Sciences under Grant No.KGZD-EW-T03the Chinese Academy of Sciences Fellowship for Young International Scientists under Grant No.2013Y1GA0004the Project funded by China Postdoctoral Science Foundation under Grant Nos.2014M550881,2015T80155.
文摘Bioluminescence tomography(BLT)is a novel opt ical molecular imaging technique that advanced the conventional planar bioluminescence imaging(BLI)into a quantifiable three-dimensional(3D)approach in preclinical living animal studies in oncology.In order to solve the inverse problem and reconstruct tumor lesions inside animal body accurately,the prior structural information is com-monly obtained from X ray computed tomography(CT).This strategy requires a complicated hybrid imaging system,extensive post imaging analysis and involvement of ionizing radiation.Moreover,the overall robustness highly depends on the fusion accuracy between the optical and structural information.Here,we present a pure optical bioluminescence tomographic(POBT)system and a novel BLT workfow based on multi-view projection acquisition and 3D surface reconstruction.This met hod can reconstruct the 3D surface of an imaging subject based on a sparse set of planar white-light and bioluminescent images,so that the prior structural information can be offered for 3D tumor lesion reconstruction without the involvement of CT.The performance of this novel technique was evaluated through the comparison with a conventional dual-modality tomo-graphic(DMT)system and a commercialized optical imaging system(IVIS Spectrum)using three breast cancer xenografts.The results revealed that the new technique offered comparable in vivo tomographic accuracy with the DMT system(P>0.05)in much shorter data analysis time.It also offered significantly better accuracy comparing with the IVIS system(P<0.04)without sacrificing too much time.
基金supported by the National Natural Science Foundation of China(Nos.81561168023,61871251,and 61871022).
文摘For fluorescence molecular tomography(FMT),image quality could be improved by incorporating a sparsity constraint.The L1 norm regularization method has been proven better than the L2 norm,like Tikhonov regularization.However,the Tikhonov method was found capable of achieving a similar quality at a high iteration cost by adopting a zeroing strategy.By studying the reason,a Tikhonov-regularization-based projecting sparsity pursuit method was proposed that reduces the iterations significantly and achieves good image quality.It was proved in phantom experiments through time-domain FMT that the method could obtain higher accuracy and less oversparsity and is more applicable for heterogeneous-target reconstruction,compared with several regularization methods implemented in this Letter.
基金the National Institutes of Health(NCI)R21 CA116102 and R21 CA125088the National Science Foundation BES-0616031,and the American Cancer Society RSG-07-005-01.
文摘There is a growing realization that cell-to-cell variations in gene expression have importantbiological consequences underlying phenotype diversity and cell fate. Although analytical toolsfor measuring gene expression, such as DNA microarrays, reverse-transcriptase PCR and in situhybridization have been widely utilized to discover the role of genetic variations in governingcellular behavior, these methods are performed in cell lysates and/or on fixed cells, and thereforelack the ability to provide comprehensive spatial-dynamic information on gene expression. Thishas invoked the recent development of molecular imaging strategies capable of illuminatingthe distribution and dynamics of RNA molecules in living cells. In this review, we describe aclass of molecular imaging probes known as molecular beacons (MBs), which have increasinglybecome the probe of choice for imaging RNA in living cells. In addition, we present the majorchallenges that can limit the ability of MBs to provide accurate measurements of RNA, anddiscuss efforts that have been made to overcome these challenges. It is envisioned that withcontinued refinement of the MB design, MBs will eventually become an indispensable tool foranalyzing gene expression in biology and medicine.
基金supported by the National Natural Science Foundation of China(Grant No.:81703466)the Outstanding Talents Research Start-up Fund of Xuzhou Medical University,China(Grant No.:RC20552107)Xuzhou Science and Technology Bureau,China(Grant No.:KC21292).
文摘Fluorescence imaging is a noninvasive and dynamic real-time imaging technique;however,it exhibits poor spatial resolution in centimeter-deep tissues because biological tissues are highly scattering media for optical radiation.The recently developed ultrasound-controlled fluorescence(UCF)imaging is a novel imaging technique that can overcome this bottleneck.Previous studies suggest that the effective contrast agent and sensitive imaging system are the two pivotal factors for generating high-resolution UCF images ex vivo and/or in vivo.Here,this review highlights the recent advances(2015e2020)in the design and synthesis of contrast agents and the improvement of imaging systems to realize high-resolution UCF imaging of deep tissues.The imaging performances of various UCF systems,including the signal-to-noise ratio,imaging resolution,and imaging depth,are specifically discussed.In addition,the challenges and prospects are highlighted.With continuously increasing research interest in this field and emerging multidisciplinary applications,UCF imaging with higher spatial resolution and larger imaging depth may be developed shortly,which is expected to have a far-reaching impact on disease surveillance and/or therapy.
文摘In this study we implemented an axillary SLN invasion model to develop highly sensitive imaging strategies enabling detection of a very small amount of tumor cells. A highly diffusible molecular probe targeting αvβ3 and αvβ5 integrins was investigated either via IV or locoregional injections. We additionally documented the potential interferences of this Near Infrared Fluorescence Probe with Blue Patente V and ICG dyes routinely used to facilitate lymph node detection during surgery. The human mammary adenocarcinoma MDA-MB-231-luc model was injected into the forepaw of nude female rats to obtain a controlled invasion of the axillary LN. Thanks to its high sensitivity, BLI was selected to achieve in vivo quantitation of tumor cells in SLNs and determine eligible animals for the study. NIRF of integrins was performed at 680 nm both in vivo and ex vivo using spectral unmixing to suppress auto-fluorescence signal and preserve sensitivity. In vivo BLI was quite reliable in estimating discrete invasion by cancer cells in the LN with thresholds of detection and quantitation of about 500 and 1500 cells respectively. For fluorescence at 680 nm, in vivo imaging is not suitable to detect micro-invasion, but ex vivo fluorescence with spectral unmixing of SLNs confirmed the presence of a tumor burden as low as 1500 cells expressing αvβ3/αvβ5 integrins. Targeting few tumor cells inside a micro-invaded sentinel lymph node by molecular probes is not sensitive enough to provide direct in vivo or peroperative imaging. At the time NIRF is performed on the excised specimen, high sensitivity imaging associated with spectral unmixing allowed such detection within less than 1 minute of examination.
基金The authors would like to thank X.Zhang,faculty of XinAoMDT Technology Co.,Ltd.,for the work of system software development.This work is supported by the National Natural Science Foundation of China under Grant Nos.81071191,60831003,30930092,30872633the Tsinghua-Yue-Yuen Medical Science Foundationthe National Basic Research Program of China(973)under Grant No.2011CB707701.
文摘We report on tests of combined positron emission tomography(PET)andfluorescence molecular tomography(FMT)imaging system for in vivo investigation on small animals.A nude mouse was inoculated with MD-MB-231 breast cancer cells which expressed redfluorescent protein(RFP).For FMT system,reflective illumination mode was adopted with full-angle data acquisition.[18F]-Fluorodeoxyglucose([18F]-FDG)was used as radioactive tracer for PET.Both data were acquired simultaneously and then reconstructed separately before fusion.Fluorescent tomography results showed exactly where the tumor was located while PET results offered more metabolic information.Results confirmed feasibility for tumor detection and showed superiority to single modality imaging.
基金supported by Ministry of Science and Technology of China under Grant No.2017YFA0205200,2016YFC0103702National Natural Science Foundation of China under Grant No.81227901,81527805‘Chinese Academy of Sciences under Grant No.GJJSTD20170004’Beijing Municipal Science&Technology Commission No.Z161100002616022by the General Financial Grant from the China Postdoctoral Science Foundation under Grant 2017 M620952.
文摘Molecular imaging(MI)is a novel imaging discipline that has been continuously developed in recent years.It combines biochemistry,multimodal imaging,biomathematics,bioinformatics,cell&molecular physiology,biophysics,and pharmacology,and it provides a new technology platform for the early diagnosis and quantitative analysis of diseases,treatment monitoring and evaluation,and the development of comprehensive physiology.Fluorescence Molecular Tomography(FMT)is a type of optical imaging modality in MI that captures the three-dimensional distribution of fluorescence within a biological tissue generated by a specific molecule of fluorescent material within a biological tissue.Compared with other optical molecular imaging methods,FMT has the characteristics of high sensitivity,low cost,and safety and reliability.It has become the research frontier and research hotspot of optical molecular imaging technology.This paper took an overview of the recent methodology advances in FMT,mainly focused on the photon propagation model of FMT based on the radiative transfer equation(RTE),and the reconstruction problem solution consist of forward problem and inverse problem.We introduce the detailed technologies utilized in reconstruction of FMT.Finally,the challenges in FMT were discussed.This survey aims at summarizing current research hotspots in methodology of FMT,fromwhich future research may benefit.
基金Supported by the National Natural Science Foundation of China(Nos.11475248 and 11105213)
文摘On May 6, 2009, the X-ray imaging and biomedical application beamline(BL13W1) at Shanghai Synchrotron Radiation Facility(SSRF) officially opened to users, with 8–72.5 ke V X-rays. The experimental station is equipped with four sets of X-ray CCD detectors of different pixel size(0.19–24 μm) for on-line phase-contrast imaging and micro-CT imaging with 0.8 μm spatial resolution and 1 ms temporal resolution. An in vivo microCT experiment for a living insect was realized in 4 s. An X-ray fluorescence detector is equipped for X-ray fluorescence mapping imaging and X-ray fluorescence micro-CT imaging with 50 μm spatial resolution. In order to meet different requirements from the users, several experimental methods, such as X-ray spiral micro-CT, Xray local micro-CT, X-ray fast micro-CT, X-ray grating-based differential micro-CT, X-ray fluorescence microCT and X-ray quantitative micro-CT have been developed, and nearly 60 papers related to those developments for this beamline have been published. Moreover, the beamline has realized the remote fast CT reconstruction,providing a great convenience for the users to process experimental data at their offices. As of August 2014,the beamline has offered the user beamtime of(23 145 h), from which 232 user papers have been published,including 151 SCI papers and 55 papers with SCI impact factor > 3. The quantity and quality of the user paper outcome keep a steady increase. Some typical user experimental results are introduced.
基金Supported by The National Research Foundation of Korea grant funded by the Korea government No. 2010-0023295the Songeui Scholar Research grant funded by the Catholic University of Korea
文摘Molecular imaging has emerged as a new discipline in gastrointestinal endoscopy.This technology encompasses modalities that can visualize disease-specific morphological or functional tissue changes based on the molecular signature of individual cells.Molecular imaging has several advantages including minimal damage to tissues,repetitive visualization,and utility for conducting quantitative analyses.Advancements in basic science coupled with endoscopy have made early detection of gastrointestinal cancer possible.Molecular imaging during gastrointestinal endoscopy requires thedevelopment of safe biomarkers and exogenous probes to detect molecular changes in cells with high specificity anda high signal-to-background ratio.Additionally,a high-resolution endoscope with an accurate wide-field viewing capability must be developed.Targeted endoscopic imaging is expected to improve early diagnosis and individual therapy of gastrointestinal cancer.
基金This work was supported in part by the Fundamental Research Funds for the Central Universities(HUST2010MS101),the NSFC(20874025)the Program for New Century Excellent Talents(NCET-07-00273)+1 种基金National Key Basic Research Program of China(2007CB310500)the National Science Foundation(NSF)Chemistry Division(CHE-0805547).
文摘Photosensitive fluorescent probes have become powerful tools in chemical biology and molecular biophysics,which are used to investigate cellular processes with high temporal and spatial resolution.Accordingly,photosensitive fluorescent probes,including photoactivatable,photoconvertible,and photoswitchable fluorophores,have been extensively developed during the past decade.The photoswitchable fluorophores have received much attention because they highlight cellular events clearly.This minireview summarizes recent advances of using reversibly photoswitchable fluorophores and their applications in innovative bioimaging.Photoswitchable fluorophores include photoswitchable fluorescent proteins,photoswitchable fluorescent organic molecules(dyes),and photoswitchable fluorescent nanoparticles.Several strategies have been developed to synthesize photoswitchable fluorophores,including engineering combination proteins,chemical synthesis,polymerization,and self-assembly.Here we concentrate on polymer nanoparticles with optically switchable emission properties:either fluorescence on/offor dualalternating-color fluorescence photoswitching.The essential mechanisms of fluorescence photoswitching enable different types of photoswitchable fluorophores to change emission intensity or wavelength(color)and thus validating the basis of the fluorescence on/offor dual-color photoswitching design.Generally the possible applications of any fluorophores are to label biological targets,followed by specific imaging.The newly developed photoswitchable fluorophores enable super-resolution fluorescence imaging because of their photosensitive emission.Finally,we summarize the important area regarding future research and development on photoswitchable fluorescent nanoparticles.
基金supported by the Natural Science Foundation of Hunan Province(2022JJ20005)National Natural Science Foundation of China(22174038,21925401,and 52221001),and Tencent Foundation.
文摘Endogenous biomolecules in cells are the basis of all life activities.Directly visualizing the structural characteristics and dynamic behaviors of cellular biomolecules is signiffcant for understanding the molecular mechanisms in various biological processes.Singlemolecule localization microscopy(SMLM)can circumvent the optical diffraction limit,achieving analysis of the ffne structures and biological processes in living cells with nanoscale resolution.However,the large size of traditional imaging probes prevents SMLM from accurately locating ffne structures and densely distributed biomolecules within cells.In recent years,nucleic acid probes have emerged as potential tools to replace conventional SMLM probes by virtue of their small size and high speciffcity.In addition,due to their programmability,nucleic acid probes with different conformations can be constructed via sequence design,further extending the application of SMLM in bioanalysis.Here,we discuss the design concepts of different conformational nucleic acid probes for SMLM and summarize the application of SMLM based on nucleic acid probes in the ffeld of biomolecules.Furthermore,we provide a summary and future perspectives of the nucleic acid probe-based SMLM technology,aiming to provide guidance for the acquisition of nanoscale information about cellular biological processes.
基金supported by National Basic Research Program of China (973 Program) (No. 2011CB707702)National Natural Science Foundation of China (Nos. 81090272, 81000632, and 30900334)+1 种基金the Shaanxi Provincial Natural Science Foundation (No. 2009JQ8018)the Fundamental Research Funds for the Central Universities
文摘A two-stage source reconstruction algorithm for bioluminescence tomography (BLT) is developed using hybrid finite element method (FEM). The proposed algorithm takes full advantages of linear and quadratic FEMs, which can be used to localize and quantify bioluminescent source accurately. In the first stage, a large permissible region is roughly determined and then iteratively evolved to reduce matrix dimension using efficient linear FEM. In the final stage, high-convergence quadratic FEM is applied to improve reconstruction result. Both numerical simulation and physical experiment are performed to evaluate the proposed algorithm. The relevant results demonstrate that quantitative reconstruction can be well achieved in terms of computation efficiency, source position, power density, and total power when compared with previous studies.
文摘Laminar optical tomography(LOT)is a mesoscopic tomographic imaging technique ranging between confocal microscopy and diffuse optical tomography(DOT).Fluorescence LOT(FLOT)provides depth-resolved molecular information with 100-200μm resolution over 2-3mm depth.In this study,we use Monte Carlo simulation and singular-value analysis(SVA)to optimize the source-detector configurations for potential enhancement of FLOT imaging performance.The effects of different design parameters,including source incidence and detector collection angles,detector number,and sampling density,are presented.The results indicate that angled incidence/detection configuration might improve the imaging resolution and depth sensitivity,especially for low-scattering medium.Increasing the number of detectors and the number of scanning steps will also result in enhanced imaging performance.We also demonstrate that the optimal imaging performance depends upon the background scattering coe±cient.Our result might provide an optimization strategy for FLOT or LOT experimental setup.
基金supported by the National Key Research and Development Program of China(Grant No.2021YFA1201203).
文摘The real-time, noninvasive, nonionizing, high spatiotemporal resolution, and flexibility characteristics of molecular fluorescence imaging provide a uniquely powerful approach to imaging and monitoring the physiology and pathophysiology of ischemic stroke. Currently, various fluorescence probes have been synthesized with the aim of improving quantitative and quantitative studies of the pathologic processes of ischemic stroke in living animals. In this review, we present an overview of current activatable fluorescence probes for the imaging and diagnosis of ischemic stroke in animal models. We categorize the probes based on their activatable signals from the biomarkers associated with ischemic stroke, and we present representative examples of their functional mechanisms. Finally, we briefly discuss future perspectives in this field.
文摘Fluorescence liftime imaging (FLIM) of modified hydrophobic bodipy dyes that act as fluorescent molecular rotors shows that the fluorescence lifetime of these probes is a function of the microviscosity of their environment. Incubating cells with these dyes, we find a punctate and continuous distribution of the dye in cells. The viscosity value obtained in what appears to be endocytotic vesicles in living cells is around 100 times higher than that of water and of cellular cytoplasm.Time-resolved fluorescence anisotropy measurements also yield rotational correlation times consistent with large microviscosity values. In this way, we successfully develop a practical and versatile approach to map the microviscosity in cells based on imaging fluorescent molecular rotors.