Recent studies in oncology have addressed the importance of detecting circulating tumor cell clusters because circulating tumor cell clusters might survive and metastasize more easily than single circulating tumor cel...Recent studies in oncology have addressed the importance of detecting circulating tumor cell clusters because circulating tumor cell clusters might survive and metastasize more easily than single circulating tumor cells.Signals with larger peak widths detected by in vivo flow cytometer(IVFC)have been used to identify cell clusters in previous studies.However,the accuracy of this criterion might be greatly degraded by variance in blood°ow and the rolling behaviors of circulating tumor cells.Here,we propose a criterion and algorithm to distinguish cell clusters from single cells.In this work,we first used area-based and volume-based models for single°uorescent cells.Simulating each model,we analyzed the corresponding morphology of IVFC signals from cell clusters.According to the Rayleigh criterion,the valley between two adjacent peak signals from two distinguishable cells should be lower than 73.5%of the peak values.A novel signal processing algorithm for IVFC was developed based on this criterion.The results showed that cell clusters can be reliably identied using our proposed algorithm.Intravital imaging was also performed to further support our algorithm.With enhanced accuracy,IVFC is a powerful tool to study circulating cell clusters.展开更多
Plankton size structure is crucial for understanding marine ecosystem dynamics and the associated biogeochemical processes.A fixation step by acid Lugol’s solution has been commonly employed to preserve plankton samp...Plankton size structure is crucial for understanding marine ecosystem dynamics and the associated biogeochemical processes.A fixation step by acid Lugol’s solution has been commonly employed to preserve plankton samples in the field.However,the acid Lugol’s solution can bias the estimation of size structure and the preserved plankton size structure can vary with time.Here,we explore the impact of sample storage time on the size-structure of the plankton community preserved by Lugol’s solution.Two short-term experiments and one long-term experiment were conducted to explore the change of plankton community size structure with the storage time:covering from a week to a month,and to nearly seven months based on particle-size data obtained by continuous Flow Cytometer and Microscope(FlowCAM)measurements.We found a linear change of plankton size with the storage time in short-term periods(less than 3 months)with a decrease of the slope but an increase of the intercept for the normalized biomass size spectrum(NBS S).However,there were opposite trends for NBSS with increasing slope but decreasing intercept after3 months.The potential causes of the distinct patterns of the NBSS parameters are addressed in terms of the interplay between particle aggregation and fragmentation.We found large changes in plankton biovolume and abundance among different size classes,which may indicate a distinct effect of acid Lugol’s solution on various plankton size classes.The mechanism driving temporal change in the size-structure of the Lugolfixed plankton community was further discussed in terms of particle aggregation and fragmentation.Finally,we emphasize that the effect of storage time should be taken into account when interpreting or comparing data of plankton community acquired from samples with various storage durations.展开更多
Metastasis is a very complicated multi-step process and accounts for the low survival rate of the cancerous patients.To metastasize,t he malignant cells must detach from the primary tumor and migrate to secondary site...Metastasis is a very complicated multi-step process and accounts for the low survival rate of the cancerous patients.To metastasize,t he malignant cells must detach from the primary tumor and migrate to secondary sites in the body through either blood or lymph circulation.Macrophages appear to be directly involved in tumor progression and metastasis.However,the role of macrophages in affecting cancer metast asis has not been fully elucidated.Here,we have utilized an emerging technique,namely in vivo flow cytometry(IVFC)to study the depletion kinetics of circulating prostate cancer cells in mice and determine how depletion of macrophages by the liposome encapsulated clodronate affects the depletion kinetics.Our results show diferent depletion kinetics of PC-3 cells between the macrophagedeficient group and the control group.The number of circulating tumor cells(CTCs)in the macrophage-deficient group decreases in a slower manner compared to the control mice group.The differences in depletion kinetics indicate that the absence of macrophages facilitates the stay of prostate cancer cells in circulation.In addition,our imaging data suggest that macrophages might be able to arrest,phagocytose and digest PC-3 cells.Therefore,phagocy tosis may mainly contribute to the de-pletion kinetic diferences.The developed methods elaborated here would be useful to study the relationship between macr ophages and tumor metastasis in small animal cancer models.展开更多
The Microchip Imaging Cytometer(MIC)is a class of integrated point-of-care detection systems based on the combination of optical microscopy and flow cytometry.MIC devices have the attributes of portability,cost-effect...The Microchip Imaging Cytometer(MIC)is a class of integrated point-of-care detection systems based on the combination of optical microscopy and flow cytometry.MIC devices have the attributes of portability,cost-effectiveness,and adaptability while providing quantitative measurements to meet the needs of laboratory testing in a variety of healthcare settings.Based on the use of microfluidic chips,MIC requires less sample and can complete sample preparation automatically.Therefore,they can provide quantitative testing results simply using a finger prick specimen.The decreased reagent consumption and reduced form factor also help improve the accessibility and affordability of healthcare services in remote and resource-limited settings.In this article,we review recent developments of the Microchip Imaging Cytometer from the following aspects:clinical applications,microfluidic chip integration,imaging optics,and image acquisition.Following,we provide an outlook of the field and remark on promising technologies that may enable significant progress in the near future.展开更多
Background: B-cell Acute lymphoblastic leukemia (B-ALL) is a neoplasm of lymphoblasts which are of B-cell lineage typically composed of small to medium sized blast cells, moderately condensed to dispersed chromatin wi...Background: B-cell Acute lymphoblastic leukemia (B-ALL) is a neoplasm of lymphoblasts which are of B-cell lineage typically composed of small to medium sized blast cells, moderately condensed to dispersed chromatin with scanty cytoplasm and inconspicuous nucleoli, involving the bone marrow and/or blood. Methods and materials: This is a prospective cross-sectional study in which 50 blood and/or bone marrow samples of newly diagnosed patients (B-ALL) were tested for immunophenotyping. All samples were prepared for surface and cytoplasmic markers including kappa and lambda human antibody for 10 minutes in dark place and then run by the Flow Cytometer. Results: 64% of the study populations were males and 36% were females. Cases were classified according to immunophenotype and the age into different subtypes and showed the following frequencies: Pro B-ALL (8%), early pre B-ALL (56%), common B-ALL (16%), Pre-B-ALL (14%) and Mature B-ALL (only 6%). Surface immunoglobulin was positive in 10% and negative in 90% of all patients, showing 100% positivity in mature B-ALL and totally negative in other subtypes. While cytoplasmic immunoglobulin was positive in 16% and negative in 84% of all patients and was positive in 100% of Pre-B-ALL and in 50% of mature B-ALL. Surface kappa was more expressed in mature B-ALL than lambda giving a ratio of 2:1, while cytoplasmic kappa:lambda was 6:1 in Pre-B-ALL. Conclusion: Kappa and lambda have important role in B-ALL classification which necessitates their presence in immunophenotyping of B-ALL.展开更多
基金the National Science Fund for Distinguished Young Scholars(Grant No.61425006)Program of Shanghai Technology Research Leader(Grant No.17XD1402200).
文摘Recent studies in oncology have addressed the importance of detecting circulating tumor cell clusters because circulating tumor cell clusters might survive and metastasize more easily than single circulating tumor cells.Signals with larger peak widths detected by in vivo flow cytometer(IVFC)have been used to identify cell clusters in previous studies.However,the accuracy of this criterion might be greatly degraded by variance in blood°ow and the rolling behaviors of circulating tumor cells.Here,we propose a criterion and algorithm to distinguish cell clusters from single cells.In this work,we first used area-based and volume-based models for single°uorescent cells.Simulating each model,we analyzed the corresponding morphology of IVFC signals from cell clusters.According to the Rayleigh criterion,the valley between two adjacent peak signals from two distinguishable cells should be lower than 73.5%of the peak values.A novel signal processing algorithm for IVFC was developed based on this criterion.The results showed that cell clusters can be reliably identied using our proposed algorithm.Intravital imaging was also performed to further support our algorithm.With enhanced accuracy,IVFC is a powerful tool to study circulating cell clusters.
基金Supported by the Guangdong Province Special Support Plan for Leading Talents(No.2019TX05H216)the Key Special Project for Introduced Talents Team of Southern Marine Science and Engineering Guangdong Laboratory(Guangzhou)(No.GML2019ZD0305)+1 种基金the National Natural Science Foundation of China(No.41906132)the Science and Technology Program of Guangzhou(No.202102021229)。
文摘Plankton size structure is crucial for understanding marine ecosystem dynamics and the associated biogeochemical processes.A fixation step by acid Lugol’s solution has been commonly employed to preserve plankton samples in the field.However,the acid Lugol’s solution can bias the estimation of size structure and the preserved plankton size structure can vary with time.Here,we explore the impact of sample storage time on the size-structure of the plankton community preserved by Lugol’s solution.Two short-term experiments and one long-term experiment were conducted to explore the change of plankton community size structure with the storage time:covering from a week to a month,and to nearly seven months based on particle-size data obtained by continuous Flow Cytometer and Microscope(FlowCAM)measurements.We found a linear change of plankton size with the storage time in short-term periods(less than 3 months)with a decrease of the slope but an increase of the intercept for the normalized biomass size spectrum(NBS S).However,there were opposite trends for NBSS with increasing slope but decreasing intercept after3 months.The potential causes of the distinct patterns of the NBSS parameters are addressed in terms of the interplay between particle aggregation and fragmentation.We found large changes in plankton biovolume and abundance among different size classes,which may indicate a distinct effect of acid Lugol’s solution on various plankton size classes.The mechanism driving temporal change in the size-structure of the Lugolfixed plankton community was further discussed in terms of particle aggregation and fragmentation.Finally,we emphasize that the effect of storage time should be taken into account when interpreting or comparing data of plankton community acquired from samples with various storage durations.
基金supported by the National Basic Research Program of China(973Program,2011CB910404 and 2012CB966800)China National Natural Science Foundation(30901432 and 30972949)+2 种基金the Chinese Ministry of Education(109056)Program for New Century Excellent Talents in University Award(NCET-08-0131)Shanghai Science and Tech-nology Committee(11DZ2211000).
文摘Metastasis is a very complicated multi-step process and accounts for the low survival rate of the cancerous patients.To metastasize,t he malignant cells must detach from the primary tumor and migrate to secondary sites in the body through either blood or lymph circulation.Macrophages appear to be directly involved in tumor progression and metastasis.However,the role of macrophages in affecting cancer metast asis has not been fully elucidated.Here,we have utilized an emerging technique,namely in vivo flow cytometry(IVFC)to study the depletion kinetics of circulating prostate cancer cells in mice and determine how depletion of macrophages by the liposome encapsulated clodronate affects the depletion kinetics.Our results show diferent depletion kinetics of PC-3 cells between the macrophagedeficient group and the control group.The number of circulating tumor cells(CTCs)in the macrophage-deficient group decreases in a slower manner compared to the control mice group.The differences in depletion kinetics indicate that the absence of macrophages facilitates the stay of prostate cancer cells in circulation.In addition,our imaging data suggest that macrophages might be able to arrest,phagocytose and digest PC-3 cells.Therefore,phagocy tosis may mainly contribute to the de-pletion kinetic diferences.The developed methods elaborated here would be useful to study the relationship between macr ophages and tumor metastasis in small animal cancer models.
基金Natural Sciences and Engineering Research Council of Canada (NSERC)Ontario Research Fund (ORF)the India-Canada Centre for Innovative Multidisciplinary Partnerships to Accelerate Community Transformation and Sustainability (IC-IM-PACTS)
文摘The Microchip Imaging Cytometer(MIC)is a class of integrated point-of-care detection systems based on the combination of optical microscopy and flow cytometry.MIC devices have the attributes of portability,cost-effectiveness,and adaptability while providing quantitative measurements to meet the needs of laboratory testing in a variety of healthcare settings.Based on the use of microfluidic chips,MIC requires less sample and can complete sample preparation automatically.Therefore,they can provide quantitative testing results simply using a finger prick specimen.The decreased reagent consumption and reduced form factor also help improve the accessibility and affordability of healthcare services in remote and resource-limited settings.In this article,we review recent developments of the Microchip Imaging Cytometer from the following aspects:clinical applications,microfluidic chip integration,imaging optics,and image acquisition.Following,we provide an outlook of the field and remark on promising technologies that may enable significant progress in the near future.
文摘Background: B-cell Acute lymphoblastic leukemia (B-ALL) is a neoplasm of lymphoblasts which are of B-cell lineage typically composed of small to medium sized blast cells, moderately condensed to dispersed chromatin with scanty cytoplasm and inconspicuous nucleoli, involving the bone marrow and/or blood. Methods and materials: This is a prospective cross-sectional study in which 50 blood and/or bone marrow samples of newly diagnosed patients (B-ALL) were tested for immunophenotyping. All samples were prepared for surface and cytoplasmic markers including kappa and lambda human antibody for 10 minutes in dark place and then run by the Flow Cytometer. Results: 64% of the study populations were males and 36% were females. Cases were classified according to immunophenotype and the age into different subtypes and showed the following frequencies: Pro B-ALL (8%), early pre B-ALL (56%), common B-ALL (16%), Pre-B-ALL (14%) and Mature B-ALL (only 6%). Surface immunoglobulin was positive in 10% and negative in 90% of all patients, showing 100% positivity in mature B-ALL and totally negative in other subtypes. While cytoplasmic immunoglobulin was positive in 16% and negative in 84% of all patients and was positive in 100% of Pre-B-ALL and in 50% of mature B-ALL. Surface kappa was more expressed in mature B-ALL than lambda giving a ratio of 2:1, while cytoplasmic kappa:lambda was 6:1 in Pre-B-ALL. Conclusion: Kappa and lambda have important role in B-ALL classification which necessitates their presence in immunophenotyping of B-ALL.
