AIM:To explore the DNA image cytometry(DNA-ICM)technique as a primary screening method for esopha-geal squamous precancerous lesions.METHODS:This study was designed as a population-based screening study.A total of 582...AIM:To explore the DNA image cytometry(DNA-ICM)technique as a primary screening method for esopha-geal squamous precancerous lesions.METHODS:This study was designed as a population-based screening study.A total of 582 local residents aged 40 years-69 years were recruited from Linzhou in Henan and Feicheng in Shandong.However,only 452 subjects had results of liquid-based cytology,DNA-ICM and pathology.The sensitivity and specificity of DNA-ICM were calculated and compared with liquid-based cytology in moderate dysplasia or worse.RESULTS:Sensitivities of DNA-ICM ranging from at least 1 to 4 aneuploid cells were 90.91%,86.36%,79.55%and 77.27%,respectively,which were better than that of liquid-based cytology(75%).Specifici-ties of DNA-ICM were 70.83%,84.07%,92.65%and 96.81%,but the specificity of liquid-based cytology was 91.91%.The sensitivity and specificity of a combination of liquid-based cytology and DNA-ICM were 84.09%and 85.78%,respectively.CONCLUSION:It is possible to use DNA-ICM tech-nique as a primary screening method for esophageal squamous precancerous lesions.展开更多
Objective： To evaluate the feasibility of DNA image cytometry （DNA-ICM） as a primary screening method for esophageal squamous cell cancer （ESCC）. Methods： A total of 5,382 local residents aged 40-69 years from t...Objective： To evaluate the feasibility of DNA image cytometry （DNA-ICM） as a primary screening method for esophageal squamous cell cancer （ESCC）. Methods： A total of 5,382 local residents aged 40-69 years from three high-risk areas in China （Linzhou in Henan province, Feicheng in Shandong province and Cixian in Hebei province） from 2008 to 2011 were recruited in this population-based screening study. And 2,526 subjects declined to receive endoscopic biopsy examination with Lugol＇s iodine staining, while 9 and 815 subjects were excluded from liquid-based cytology and DNA-ICM test respectively due to slide quality. Finally, 2,856, 5,373 and 4,567 subjects were enrolled in the analysis for endoscopic biopsy examination, liquid-based cytology and DNA-ICM test, respectively. Sensitivity （SE）, specificity （SP）, negative predictive values （NPV） and positive predictive values （PPV） as well as their 95% confidence intervals （95% CI） for DNA-ICM, liquid-based cytology and the combination of the two methods were calculated. Receiver operating characteristic （ROC） curves were applied to determine the cutoff point of DNA-ICM for esophageal cancer. Results： DNA-ICM results were significantly correlative with esophageal cancer and precancer lesions （X2= 18.016, P〈0.001）. The cutoff points were 5,802, 5,803 and 8,002 based on dissimilar pathological types of low grade intraepithelial neoplasia （LGIN）, high grade intraepithelial neoplasia （HGIN）, and ESCC, respectively, and 5,803 was chosen in this study considering the SE and SP. The SE, SP, PPV, NPV of DNA-ICM test （cutoff point 5,803） combined with liquid-based cytology [threshold atypical squamous cells of undetermined significance （ASCUS）] were separately 72.1% （95% CI： 70.3%-73.9%）, 43.3% （95% CI. 41.3%-45.3%）, 22.8% （95% CI： 21.1%-24.5%） and 87.0% （95% CI： 85.7%-88.3%） for LGIN, 85.7% （95% CI： 84.3%-87.1%）, 41.3% （95% CI： 39.3%-43.3%）, 4.6% （95% CI： 3.8%-5.4%） and 98.9% （95% CI： 98.5%-99.3%） for HGIN, and 96.0% （95% CI： 95.2%-96.8%）, 40.8% （95% CI： 38.8%-42.8%）, 1.7% （95% CI： 1.2%-2.2%） and 99.9% （95% CI： 99.8%-100.0%） for ESCC. Conclusions： It is possible to use DNA-ICM test as a primary screening method before endoscopic screening for esophageal cancer.展开更多
To evaluate the value of detecton of DNA aneuploidy in exfoliated airway epithelia cells of sputum specimens by the automated image cytometry for the identification of lung cancer, 100 patients were divided into pat...To evaluate the value of detecton of DNA aneuploidy in exfoliated airway epithelia cells of sputum specimens by the automated image cytometry for the identification of lung cancer, 100 patients were divided into patient group (50 patients with lung cancer) and control group (30 patients with tuberculosis and 20 healthy people). Sputum was obtained for the quantitative analysis of DNA content of exfoliated airway epithelial cells with the automated image cytometry, together with the examinations of brush cytology and conventional sputum cytology. Our results showed that DNA aneuploidy (DI>2.5 or 5c) was found in 20 out of 50 sputum samples of lung cancer, 1 out of 30 sputum samples from tuberculosis patients, and none of 20 sputum samples from healthy people. The positive rates of conventional sputum cytology and brush cytology were 16 % and 32 %, which was lower than that of DNA aneuploidy detection by the automated image cytometry (P<0.01,P>05). Our study showed that automated image cytometry, which uses DNA aneuploidy as a marker for tumor, can detect the malignant cells in sputum samples of lung cancer and it is a sensitive and specific method serving as a complement for the diagnosis of lung cancer.展开更多
Objective:To elucidate whether DNA aneuploidy was an independent discriminator for carcinoma within oral potentially malignant disorders(OPMDs),and further establish and validate a risk model based on DNA aneuploidy f...Objective:To elucidate whether DNA aneuploidy was an independent discriminator for carcinoma within oral potentially malignant disorders(OPMDs),and further establish and validate a risk model based on DNA aneuploidy for the detection of oral cancer.Methods:A total of 810 consecutive patients with OPMD were prospectively enrolled from March 2013 to December 2018,and divided into a training set(n=608)and a test set(n=202).Brushing and biopsy samples from each patient were processed by DNADNA image cytometry and histopathological examination,respectively.Results:DNA aneuploidy of an outside DNA index≥3.5 in OPMD was an independent marker strongly associated with malignant risk[adjusted odds ratio:13.04;95%confidence interval(CI):5.46-31.14].In the training and test sets,the area under the curve(AUC)was 0.87(95%CI:0.82-0.91)and 0.77(95%CI:0.57-0.97),respectively,for detecting carcinoma in OPMD patients.The independent risk factors of lateral/ventral tongue and non-homogenous type combined with a risk model built with a multivariate logistic regression revealed a more favorable diagnostic efficacy associated with the training set(AUC:0.93;95%CI:0.91-0.96)and test set(AUC:0.94;95%CI:0.90-0.98).The sensitivity and specificity of carcinoma detection within OPMD was improved to 100%and 88.1%,respectively.Conclusions:This large-scale diagnostic study established a risk model based on DNA aneuploidy that consisted of a noninvasive strategy with lateral/ventral tongue and non-homogenous features.The results showed favorable diagnostic efficacy for detecting carcinoma within OPMD,irrespective of the clinical and pathological diagnoses of OPMD.Multicenter validation and longitudinal studies are warranted to evaluate community practices and clinical applications.展开更多
Extranodal NK/T cell lymphoma(ENKTL)poses significant challenges in efficient treatment processes due to its aggressive nature and high recurrence rates.There is a critical need to develop a robust statistical model t...Extranodal NK/T cell lymphoma(ENKTL)poses significant challenges in efficient treatment processes due to its aggressive nature and high recurrence rates.There is a critical need to develop a robust statistical model to predict treatment efficacy by dynamically quantifying biomarkers tailored to various stages of lymphoma.Recent analytics such as sequencing and microbiome tests have only been utilized to understand lymphoma progression and treatment response in clinical settings.However,these methods are limited by their quantitative analysis capabilities,long turnaround times,and lack of single-cell resolution,which are essential for understanding the heterogeneous nature of lymphoma.In this study,we developed a deep learning-enhanced image cytometry(DLIC)to investigate biophysical heterogeneities in peripheral blood mononuclear cells(PBMCs)from newly diagnosed(ND)ENKTL patients.We established a substantial cohort of 23 ND ENKTL patients,categorizing them into interim of treatment(n=21)and end of treatment(n=19)stages along their serial treatment timelines.Using a basic optical microscope and a commercial microchip,we analyzed over 270,000 single PBMCs in high-throughput,profiling their size,eccentricity,and refractive index in a completely label-free and quantified manner through Al-based nanophotonic computation.We observed distinct heterogeneity variations in these three biophysical indicators across treatment stages and relapse statuses,revealing solid mechanistic correlations among the phenotypes.We established a three-dimensional single-cell distribution map for ENKTL patients and created a standard for quantifying the change in occupational volume.Leveraging this extensive database,DLIC offers on-site analytics in clinical settings,facilitating treatment assessment and prognosis prediction through label-free biophysical analysis of patient PBMCs,extracted directly without additional sample preparation.展开更多
Imaging mass cytometry(IMC)enables the high-resolution spatial profiling of tumor microenvironment,but its clinical utility for prospective prediction remains underdeveloped.In this study,we integrated IMC into a clin...Imaging mass cytometry(IMC)enables the high-resolution spatial profiling of tumor microenvironment,but its clinical utility for prospective prediction remains underdeveloped.In this study,we integrated IMC into a clinical trial of hepatocellular carcinoma(HCC)patients undergoing combination therapy with programmed death-1 blockade and transarterial chemoembolization.We analyzed 281 regions of interest from 43 patients using a custom 40-marker IMC panel and developed a novel superpixel-based graph attention network,IMCSGAT,to model spatial cell interactions within the tumor microenvironment.IMCSGAT enabled accurate multitask prediction of key clinical features,including Barcelona Clinic Liver Cancer stage,trabecular histologic subtype,and treatment response.Compared to state-of-the-art methods,IMCSGAT achieved superior performance across all classification tasks.Spatial interaction analysis revealed that resident macrophage-centered interactions,particularly those with NK and T cells,were enriched in responders and predictive of therapeutic outcome.These findings were validated in a murine HCC model,reinforcing the role of innate immune architecture in shaping the treatment response.This study establishes IMCSGAT as a powerful spatial learning framework for high-dimensional IMC data,with potential applications in clinical outcome prediction and personalized therapy design for HCC.Our results provide a blueprint for the broader use of spatial analytics in precision oncology.展开更多
As the gold-standard method for single-cell analysis,flow cytometry enables high-throughput and multiple-parameter characterization of individual biological cells.This review highlights the demands for clinical flow c...As the gold-standard method for single-cell analysis,flow cytometry enables high-throughput and multiple-parameter characterization of individual biological cells.This review highlights the demands for clinical flow cytometry in laboratory hematology(e.g.,diagnoses of minimal residual disease and various types of leukemia),summarizes state-of-the-art clinical flow cytometers(e.g.,FACSLyricTMby Becton Dickinson,DxFLEX by Beckman Coulter),then considers innovative technical improvements in flow cytometry(including quantitative,spectral,and imaging approaches)to address the limitations of clinical flow cytometry in hematology diagnosis.Finally,driven by these clinical demands,future developments in clinical flow cytometry are suggested.展开更多
Gaining a full understanding of the mechanisms of action of natural products as therapeutic agents includes observing the effects of natural products on cellular morphology,because abnormal cellular morphology is an i...Gaining a full understanding of the mechanisms of action of natural products as therapeutic agents includes observing the effects of natural products on cellular morphology,because abnormal cellular morphology is an important aspect of cellular transformations that occur as part of disease states.In this study a set of natural products was examined in search of small molecules that influence the cylindrical morphology of fission yeast Schizosaccharomyces pombe.Imaging flow cytometry of large populations of S.pombe exposed to natural products captured cell images and revealed changes in mean length and aspect ratio of cells.Several natural products were found to alter S.pombe’s morphology relative to control,in terms of elongating cells,shrinking them,or making them more round.These results may facilitate future investigations into methods by which cells establish and maintain specific shapes.展开更多
In the tumor microenvironment(TME),various types of immune cells interact with each other and with cancer cells,playing critical roles in cancer progression and treatment[1].Numerous studies have reported that the inf...In the tumor microenvironment(TME),various types of immune cells interact with each other and with cancer cells,playing critical roles in cancer progression and treatment[1].Numerous studies have reported that the infiltration levels of specific immune cells are associated with patient prognosis and response to immunotherapies[2,3].展开更多
Imaging flow cytometry(IFC)combines the imaging capabilities of microscopy with the high throughput of flow cytometry,offering a promising solution for high-precision and high-throughput cell analysis in fields such a...Imaging flow cytometry(IFC)combines the imaging capabilities of microscopy with the high throughput of flow cytometry,offering a promising solution for high-precision and high-throughput cell analysis in fields such as biomedicine,green energy,and environmental monitoring.However,due to limitations in imaging framerate and realtime data processing,the real-time throughput of existing IFC systems has been restricted to approximately 1000-10,000 events per second(eps),which is insufficient for large-scale cell analysis.In this work,we demonstrate IFC with real-time throughput exceeding 1,000,000 eps by integrating optical time-stretch(OTS)imaging,microfluidic-based cell manipulation,and online image processing.Cells flowing at speeds up to 15 m/s are clearly imaged with a spatial resolution of 780 nm,and images of each individual cell are captured,stored,and analyzed.The capabilities and performance of our system are validated through the identification of malignancies in clinical colorectal samples.This work sets a new record for throughput in imaging flow cytometry,and we believe it has the potential to revolutionize cell analysis by enabling highly efficient,accurate,and intelligent measurement.展开更多
Mass cytometry(cytometry by time-of-flight(CyTOF))and imaging mass cytometry(IMC)are transformative technologies that combine flow cytometry principles with time-of-flight mass spectrometry(TOF-MS).By employing metal ...Mass cytometry(cytometry by time-of-flight(CyTOF))and imaging mass cytometry(IMC)are transformative technologies that combine flow cytometry principles with time-of-flight mass spectrometry(TOF-MS).By employing metal isotope-tagged antibodies instead of fluorophores,these techniques overcome spectral overlap limitations and enable high-dimensional,compensation-free analysis of complex biological systems at single-cell resolution.The performance of CyTOF and IMC critically depends on advanced nanomaterials labeled with stable metal isotopes,which are essential for improving sensitivity and multiplexing capacity.This review systematically discusses the design principles,synthesis methods,and functionalization strategies of mass-tagged nanomaterials tailored for CyTOF(e.g.,cell suspension analysis)and IMC(e.g.,spatial proteomics of tissue sections).We highlight their impactful applications in biomedicine,including proteomics,immunology,oncology,and neuroscience,emphasizing their roles in disease diagnosis,targeted drug development,and singlecell analysis.Despite these advancements,challenges such as nanomaterial biocompatibility,clinical scalability,and artificial intelligence(AI)-driven design are discussed,providing a roadmap for future research in personalized medicine and theranostics.展开更多
Mechanical measurements of cells can provide unique insights into cell state and disease processes.The overall mechanical properties of cells can be heavily affected by the stiffest organelle,the nucleus.However,it is...Mechanical measurements of cells can provide unique insights into cell state and disease processes.The overall mechanical properties of cells can be heavily affected by the stiffest organelle,the nucleus.However,it is challenging to fully characterize internal nuclear structures in most cell mechanical measurement platforms.Here,we demonstrate single-cell deformability measurements of whole cells and stained nuclei in a fluorescence imaging flow cytometry platform.We also introduce bending energy derived metrics as a way to normalize measurements of cytoskeletal cortex and nuclear shape changes of cells and demonstrate the utility of relative deformability distributions to characterize populations of cells.We apply the platform to measure changes in cell biophysical properties during the process of NETosis,whereby neutrophils undergo drastic nuclear restructuring.We characterize cell size,deformability,and nuclear structure changes and their correlations in thousands of neutrophils undergoing NETosis,a process implicated in development of critical disease states,such as sepsis.This platform can aid in understanding heterogeneity in deformability in cell populations and how this may be influenced by nuclear or internal structure changes.展开更多
Single-pixel imaging （SPI） technology has garnered great interest within the last decade because of its ability to record high-resolution images using a single-pixel detector. It has been applied to diverse fields, ...Single-pixel imaging （SPI） technology has garnered great interest within the last decade because of its ability to record high-resolution images using a single-pixel detector. It has been applied to diverse fields, such as magnetic resonance imaging （MRI）, aerospace remote sensing, terahertz photography, and hyperspectral imaging. Compared with conventional silicon-based cameras, single-pixel cameras （SPCs） can achieve image compression and operate over a much broader spectral range. However, the imaging speed of SPCs is governed by the response time of digital mieromirror devices （DMDs） and the amount of com- pression of acquired images, leading to low （ms-level） temporal resolution. Consequently, it is particularly challenging for SPCs to investigate fast dynamic phenomena, which is required commonly in microscopy. Recently, a unique approach based on photonic time stretch （PTS） to achieve high-speed SPI has been reported. It achieves a frame rate far beyond that can be reached with conventional SPCs. In this paper, we first introduce the principles and applications of the PTS technique. Then the basic archi- tecture of the high-speed SPI system is presented, and an imaging flow cytometer with high speed and high throughput is demonstrated experimentally. Finally, the limitations and potential applications of high-speed SPI are discussed.展开更多
基金Supported by Grants from the Ministry of Health of China,No.200902002-8Grants from Cancer Institute/Hospital Chinese Academy of Medical Sciences and Peking Union Medical College,No.2009YF50
文摘AIM:To explore the DNA image cytometry(DNA-ICM)technique as a primary screening method for esopha-geal squamous precancerous lesions.METHODS:This study was designed as a population-based screening study.A total of 582 local residents aged 40 years-69 years were recruited from Linzhou in Henan and Feicheng in Shandong.However,only 452 subjects had results of liquid-based cytology,DNA-ICM and pathology.The sensitivity and specificity of DNA-ICM were calculated and compared with liquid-based cytology in moderate dysplasia or worse.RESULTS:Sensitivities of DNA-ICM ranging from at least 1 to 4 aneuploid cells were 90.91%,86.36%,79.55%and 77.27%,respectively,which were better than that of liquid-based cytology(75%).Specifici-ties of DNA-ICM were 70.83%,84.07%,92.65%and 96.81%,but the specificity of liquid-based cytology was 91.91%.The sensitivity and specificity of a combination of liquid-based cytology and DNA-ICM were 84.09%and 85.78%,respectively.CONCLUSION:It is possible to use DNA-ICM tech-nique as a primary screening method for esophageal squamous precancerous lesions.
基金granted by the National Natural Science Foundation of China (No.81241091)
文摘Objective： To evaluate the feasibility of DNA image cytometry （DNA-ICM） as a primary screening method for esophageal squamous cell cancer （ESCC）. Methods： A total of 5,382 local residents aged 40-69 years from three high-risk areas in China （Linzhou in Henan province, Feicheng in Shandong province and Cixian in Hebei province） from 2008 to 2011 were recruited in this population-based screening study. And 2,526 subjects declined to receive endoscopic biopsy examination with Lugol＇s iodine staining, while 9 and 815 subjects were excluded from liquid-based cytology and DNA-ICM test respectively due to slide quality. Finally, 2,856, 5,373 and 4,567 subjects were enrolled in the analysis for endoscopic biopsy examination, liquid-based cytology and DNA-ICM test, respectively. Sensitivity （SE）, specificity （SP）, negative predictive values （NPV） and positive predictive values （PPV） as well as their 95% confidence intervals （95% CI） for DNA-ICM, liquid-based cytology and the combination of the two methods were calculated. Receiver operating characteristic （ROC） curves were applied to determine the cutoff point of DNA-ICM for esophageal cancer. Results： DNA-ICM results were significantly correlative with esophageal cancer and precancer lesions （X2= 18.016, P〈0.001）. The cutoff points were 5,802, 5,803 and 8,002 based on dissimilar pathological types of low grade intraepithelial neoplasia （LGIN）, high grade intraepithelial neoplasia （HGIN）, and ESCC, respectively, and 5,803 was chosen in this study considering the SE and SP. The SE, SP, PPV, NPV of DNA-ICM test （cutoff point 5,803） combined with liquid-based cytology [threshold atypical squamous cells of undetermined significance （ASCUS）] were separately 72.1% （95% CI： 70.3%-73.9%）, 43.3% （95% CI. 41.3%-45.3%）, 22.8% （95% CI： 21.1%-24.5%） and 87.0% （95% CI： 85.7%-88.3%） for LGIN, 85.7% （95% CI： 84.3%-87.1%）, 41.3% （95% CI： 39.3%-43.3%）, 4.6% （95% CI： 3.8%-5.4%） and 98.9% （95% CI： 98.5%-99.3%） for HGIN, and 96.0% （95% CI： 95.2%-96.8%）, 40.8% （95% CI： 38.8%-42.8%）, 1.7% （95% CI： 1.2%-2.2%） and 99.9% （95% CI： 99.8%-100.0%） for ESCC. Conclusions： It is possible to use DNA-ICM test as a primary screening method before endoscopic screening for esophageal cancer.
文摘To evaluate the value of detecton of DNA aneuploidy in exfoliated airway epithelia cells of sputum specimens by the automated image cytometry for the identification of lung cancer, 100 patients were divided into patient group (50 patients with lung cancer) and control group (30 patients with tuberculosis and 20 healthy people). Sputum was obtained for the quantitative analysis of DNA content of exfoliated airway epithelial cells with the automated image cytometry, together with the examinations of brush cytology and conventional sputum cytology. Our results showed that DNA aneuploidy (DI>2.5 or 5c) was found in 20 out of 50 sputum samples of lung cancer, 1 out of 30 sputum samples from tuberculosis patients, and none of 20 sputum samples from healthy people. The positive rates of conventional sputum cytology and brush cytology were 16 % and 32 %, which was lower than that of DNA aneuploidy detection by the automated image cytometry (P<0.01,P>05). Our study showed that automated image cytometry, which uses DNA aneuploidy as a marker for tumor, can detect the malignant cells in sputum samples of lung cancer and it is a sensitive and specific method serving as a complement for the diagnosis of lung cancer.
基金supported by the National Natural Science Foundation of China(Grant No.82074502)the Science and Technology Commission of Shanghai Municipality(Grant No.20Y11903700)+3 种基金the Shanghai Hospital Development Center(Grant No.SHDC2020CR4082)the Shanghai Municipal Health Committee(Grant No.202040457)the Innovative Research Team of High-level Local Universities in Shanghai(Grant No.SSMU-ZDCX20180901)the SHIPM-mu Fund from the Shanghai Institute of Precision Medicine(Grant No.JC201807)。
文摘Objective:To elucidate whether DNA aneuploidy was an independent discriminator for carcinoma within oral potentially malignant disorders(OPMDs),and further establish and validate a risk model based on DNA aneuploidy for the detection of oral cancer.Methods:A total of 810 consecutive patients with OPMD were prospectively enrolled from March 2013 to December 2018,and divided into a training set(n=608)and a test set(n=202).Brushing and biopsy samples from each patient were processed by DNADNA image cytometry and histopathological examination,respectively.Results:DNA aneuploidy of an outside DNA index≥3.5 in OPMD was an independent marker strongly associated with malignant risk[adjusted odds ratio:13.04;95%confidence interval(CI):5.46-31.14].In the training and test sets,the area under the curve(AUC)was 0.87(95%CI:0.82-0.91)and 0.77(95%CI:0.57-0.97),respectively,for detecting carcinoma in OPMD patients.The independent risk factors of lateral/ventral tongue and non-homogenous type combined with a risk model built with a multivariate logistic regression revealed a more favorable diagnostic efficacy associated with the training set(AUC:0.93;95%CI:0.91-0.96)and test set(AUC:0.94;95%CI:0.90-0.98).The sensitivity and specificity of carcinoma detection within OPMD was improved to 100%and 88.1%,respectively.Conclusions:This large-scale diagnostic study established a risk model based on DNA aneuploidy that consisted of a noninvasive strategy with lateral/ventral tongue and non-homogenous features.The results showed favorable diagnostic efficacy for detecting carcinoma within OPMD,irrespective of the clinical and pathological diagnoses of OPMD.Multicenter validation and longitudinal studies are warranted to evaluate community practices and clinical applications.
基金Korea Health Industry Development Institute,Grant/Award Number:HR20C0025National Research Foundation of Korea,Grant/Award Numbers:RS-2023-00211580,RS-2023-00222838。
文摘Extranodal NK/T cell lymphoma(ENKTL)poses significant challenges in efficient treatment processes due to its aggressive nature and high recurrence rates.There is a critical need to develop a robust statistical model to predict treatment efficacy by dynamically quantifying biomarkers tailored to various stages of lymphoma.Recent analytics such as sequencing and microbiome tests have only been utilized to understand lymphoma progression and treatment response in clinical settings.However,these methods are limited by their quantitative analysis capabilities,long turnaround times,and lack of single-cell resolution,which are essential for understanding the heterogeneous nature of lymphoma.In this study,we developed a deep learning-enhanced image cytometry(DLIC)to investigate biophysical heterogeneities in peripheral blood mononuclear cells(PBMCs)from newly diagnosed(ND)ENKTL patients.We established a substantial cohort of 23 ND ENKTL patients,categorizing them into interim of treatment(n=21)and end of treatment(n=19)stages along their serial treatment timelines.Using a basic optical microscope and a commercial microchip,we analyzed over 270,000 single PBMCs in high-throughput,profiling their size,eccentricity,and refractive index in a completely label-free and quantified manner through Al-based nanophotonic computation.We observed distinct heterogeneity variations in these three biophysical indicators across treatment stages and relapse statuses,revealing solid mechanistic correlations among the phenotypes.We established a three-dimensional single-cell distribution map for ENKTL patients and created a standard for quantifying the change in occupational volume.Leveraging this extensive database,DLIC offers on-site analytics in clinical settings,facilitating treatment assessment and prognosis prediction through label-free biophysical analysis of patient PBMCs,extracted directly without additional sample preparation.
基金supported by the National Natural Science Foundation of China(grant 62136004 to D.Z.,grant 62272226 to W.S.,and grant 82173078 to J.S.)the National Key Research and Development Program of China(grant 2019YFA0803000 to J.S.)+1 种基金the Excellent Youth Foundation of Zhejiang Scientific(grant R22H1610037 to J.S.)the Zhejiang Provincial Natural Science Foundation(grant 2022C03037 to J.S.).
文摘Imaging mass cytometry(IMC)enables the high-resolution spatial profiling of tumor microenvironment,but its clinical utility for prospective prediction remains underdeveloped.In this study,we integrated IMC into a clinical trial of hepatocellular carcinoma(HCC)patients undergoing combination therapy with programmed death-1 blockade and transarterial chemoembolization.We analyzed 281 regions of interest from 43 patients using a custom 40-marker IMC panel and developed a novel superpixel-based graph attention network,IMCSGAT,to model spatial cell interactions within the tumor microenvironment.IMCSGAT enabled accurate multitask prediction of key clinical features,including Barcelona Clinic Liver Cancer stage,trabecular histologic subtype,and treatment response.Compared to state-of-the-art methods,IMCSGAT achieved superior performance across all classification tasks.Spatial interaction analysis revealed that resident macrophage-centered interactions,particularly those with NK and T cells,were enriched in responders and predictive of therapeutic outcome.These findings were validated in a murine HCC model,reinforcing the role of innate immune architecture in shaping the treatment response.This study establishes IMCSGAT as a powerful spatial learning framework for high-dimensional IMC data,with potential applications in clinical outcome prediction and personalized therapy design for HCC.Our results provide a blueprint for the broader use of spatial analytics in precision oncology.
基金financial support of the National Natural Science Foundation of China(Grant Nos.61922079,61825107,and 62121003)the Chinese Academy of Sciences(Grant Nos.GJJSTD20210004 and Y201927)the National Key Research and Development Program of China(Grant No.2021YFC2500300).
文摘As the gold-standard method for single-cell analysis,flow cytometry enables high-throughput and multiple-parameter characterization of individual biological cells.This review highlights the demands for clinical flow cytometry in laboratory hematology(e.g.,diagnoses of minimal residual disease and various types of leukemia),summarizes state-of-the-art clinical flow cytometers(e.g.,FACSLyricTMby Becton Dickinson,DxFLEX by Beckman Coulter),then considers innovative technical improvements in flow cytometry(including quantitative,spectral,and imaging approaches)to address the limitations of clinical flow cytometry in hematology diagnosis.Finally,driven by these clinical demands,future developments in clinical flow cytometry are suggested.
基金financial support from the University of North Florida.
文摘Gaining a full understanding of the mechanisms of action of natural products as therapeutic agents includes observing the effects of natural products on cellular morphology,because abnormal cellular morphology is an important aspect of cellular transformations that occur as part of disease states.In this study a set of natural products was examined in search of small molecules that influence the cylindrical morphology of fission yeast Schizosaccharomyces pombe.Imaging flow cytometry of large populations of S.pombe exposed to natural products captured cell images and revealed changes in mean length and aspect ratio of cells.Several natural products were found to alter S.pombe’s morphology relative to control,in terms of elongating cells,shrinking them,or making them more round.These results may facilitate future investigations into methods by which cells establish and maintain specific shapes.
基金supported by the Cancer Prevention Research Institute of Texas(CPRIT)(RR180061)the National Cancer Institute of the National Institute of Health(1R01CA269764).
文摘In the tumor microenvironment(TME),various types of immune cells interact with each other and with cancer cells,playing critical roles in cancer progression and treatment[1].Numerous studies have reported that the infiltration levels of specific immune cells are associated with patient prognosis and response to immunotherapies[2,3].
基金supported by the National Key R&D Program of China(2023YFF0723300)National Natural Science Foundation of China(62475198,62075200,12374295)+8 种基金Fundamental Research Funds for the Central Universities(2042024kf0003,2042024kf1010,2042023kf0105)Hubei Provincial Natural Science Foundation of China(2023AFB133)Jiangsu Science and Technology Program(BK20221257)Shenzhen Science and Technology Program(JCYJ20220530140601003,JCYJ20230807090207014)Translational Medicine and Multidisciplinary Research Project of Zhongnan Hospital of Wuhan University(ZNJC202217,ZNJC202232)The Interdisciplinary Innovative Talents Foundation from Renmin Hospital of Wuhan University(JCRCYR-2022-006)Hubei Province Young Science and Technology Talent Morning Hight Lift Project(202319)The Fund of National Key Laboratory of Plasma Physics(6142A04230201)We gratefully acknowledge Serendipity Lab for facilitating collaboration opportunities.
文摘Imaging flow cytometry(IFC)combines the imaging capabilities of microscopy with the high throughput of flow cytometry,offering a promising solution for high-precision and high-throughput cell analysis in fields such as biomedicine,green energy,and environmental monitoring.However,due to limitations in imaging framerate and realtime data processing,the real-time throughput of existing IFC systems has been restricted to approximately 1000-10,000 events per second(eps),which is insufficient for large-scale cell analysis.In this work,we demonstrate IFC with real-time throughput exceeding 1,000,000 eps by integrating optical time-stretch(OTS)imaging,microfluidic-based cell manipulation,and online image processing.Cells flowing at speeds up to 15 m/s are clearly imaged with a spatial resolution of 780 nm,and images of each individual cell are captured,stored,and analyzed.The capabilities and performance of our system are validated through the identification of malignancies in clinical colorectal samples.This work sets a new record for throughput in imaging flow cytometry,and we believe it has the potential to revolutionize cell analysis by enabling highly efficient,accurate,and intelligent measurement.
基金supported by National Natural Science Foundation of China(T2122002,82361148715,22077079,82204104)National Key R&D Program of China(2022YFC2601700,2022YFF0710202 and 2022YFA1104200)+4 种基金Princess Nourah bint Abdulrahman University Researchers Supporting Project number(PNURSP2024R122)Shanghai Municipal Science and Technology Projects(22Z510202478)Shanghai Municipal Education Commission Projects(21SG10,ZXWH1082101)Shanghai Jiao Tong University Projects(YG2021ZD19)Shanghai University of Medicine&Health Sciences Project(AMSCP-24-07-01).
文摘Mass cytometry(cytometry by time-of-flight(CyTOF))and imaging mass cytometry(IMC)are transformative technologies that combine flow cytometry principles with time-of-flight mass spectrometry(TOF-MS).By employing metal isotope-tagged antibodies instead of fluorophores,these techniques overcome spectral overlap limitations and enable high-dimensional,compensation-free analysis of complex biological systems at single-cell resolution.The performance of CyTOF and IMC critically depends on advanced nanomaterials labeled with stable metal isotopes,which are essential for improving sensitivity and multiplexing capacity.This review systematically discusses the design principles,synthesis methods,and functionalization strategies of mass-tagged nanomaterials tailored for CyTOF(e.g.,cell suspension analysis)and IMC(e.g.,spatial proteomics of tissue sections).We highlight their impactful applications in biomedicine,including proteomics,immunology,oncology,and neuroscience,emphasizing their roles in disease diagnosis,targeted drug development,and singlecell analysis.Despite these advancements,challenges such as nanomaterial biocompatibility,clinical scalability,and artificial intelligence(AI)-driven design are discussed,providing a roadmap for future research in personalized medicine and theranostics.
基金supported by ImPACT Program of the Council of Science,Technology and Innovation(Cabinet office,Government of Japan)and the Presidential Early Career Award for Scientists and Engineers to D.D.(N00014-16-1-2997).
文摘Mechanical measurements of cells can provide unique insights into cell state and disease processes.The overall mechanical properties of cells can be heavily affected by the stiffest organelle,the nucleus.However,it is challenging to fully characterize internal nuclear structures in most cell mechanical measurement platforms.Here,we demonstrate single-cell deformability measurements of whole cells and stained nuclei in a fluorescence imaging flow cytometry platform.We also introduce bending energy derived metrics as a way to normalize measurements of cytoskeletal cortex and nuclear shape changes of cells and demonstrate the utility of relative deformability distributions to characterize populations of cells.We apply the platform to measure changes in cell biophysical properties during the process of NETosis,whereby neutrophils undergo drastic nuclear restructuring.We characterize cell size,deformability,and nuclear structure changes and their correlations in thousands of neutrophils undergoing NETosis,a process implicated in development of critical disease states,such as sepsis.This platform can aid in understanding heterogeneity in deformability in cell populations and how this may be influenced by nuclear or internal structure changes.
基金Project supported by the National Natural Science Foundation of China （Nos. 61771284 and 61322113）
文摘Single-pixel imaging （SPI） technology has garnered great interest within the last decade because of its ability to record high-resolution images using a single-pixel detector. It has been applied to diverse fields, such as magnetic resonance imaging （MRI）, aerospace remote sensing, terahertz photography, and hyperspectral imaging. Compared with conventional silicon-based cameras, single-pixel cameras （SPCs） can achieve image compression and operate over a much broader spectral range. However, the imaging speed of SPCs is governed by the response time of digital mieromirror devices （DMDs） and the amount of com- pression of acquired images, leading to low （ms-level） temporal resolution. Consequently, it is particularly challenging for SPCs to investigate fast dynamic phenomena, which is required commonly in microscopy. Recently, a unique approach based on photonic time stretch （PTS） to achieve high-speed SPI has been reported. It achieves a frame rate far beyond that can be reached with conventional SPCs. In this paper, we first introduce the principles and applications of the PTS technique. Then the basic archi- tecture of the high-speed SPI system is presented, and an imaging flow cytometer with high speed and high throughput is demonstrated experimentally. Finally, the limitations and potential applications of high-speed SPI are discussed.
