Methods in cell science : an official journal of the Society for In Vitro Biology最新文献

Chromosome Painting 染色体涂染

Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2020-02-02 DOI: 10.1007/978-94-010-0330-8

A. Sharma, A. Sharma

引用次数: 39

Data reproducibility in fluorescence image analysis. 荧光图像分析中的数据再现性。

Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1007/s11022-004-2383-4

Catherine Souchier, Christine Brisson, Bernadette Batteux, Michel Robert-Nicoud, Paul-André Bryon

{"title":"Data reproducibility in fluorescence image analysis.","authors":"Catherine Souchier, Christine Brisson, Bernadette Batteux, Michel Robert-Nicoud, Paul-André Bryon","doi":"10.1007/s11022-004-2383-4","DOIUrl":"https://doi.org/10.1007/s11022-004-2383-4","url":null,"abstract":"Fluorescence image analysis provides quantitative data on fluorescence in situ hybridization signals (FISH), immunofluorescence labelings, Green Fluorescent Protein (GFP) expression and microarrays. It is a valuable tool for decision making in the fields of biology and medicine. The aim of this study was to evaluate the reproducibility of fluorescence intensity measurements and standardization when acquisitions are performed under various but well defined conditions. Fluorescent intensity of standard beads (Inspeck series, Molecular Probes) was repeatedly measured using an image analyzer and automated procedures. Images were acquired using several integration times and neutral filter sets. A standardization procedure was used for expressing the data in a same unit: data were multiplied by the light attenuation factor and were divided by the CCD integration times. Results show that 1) standardization is possible 2) accurate and reliable fluorescence measurements can be obtained and 3) specimens showing large differences in fluorescence intensity can be objectively compared. Moreover fluorescent test slides including fluorochrome solutions and altuglas slides were tested for shading correction and as overall test systems.","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"25 3-4","pages":"195-200"},"PeriodicalIF":0.0,"publicationDate":"2003-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s11022-004-2383-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25203827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10

A reproducible modified method for direct preparation of chorionic villi cytogenetic analysis. 直接制备绒毛膜绒毛细胞遗传学分析的可重复性改进方法。

Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1007/s11022-004-6830-z

Shashirekha Shetty, Alka Gogate, Sharad Gogate, Paul Malet

引用次数: 2

Whole-cell patch-clamp recordings from identified spinal neurons in the zebrafish embryo. 斑马鱼胚胎中鉴定的脊髓神经元的全细胞膜片钳记录。

Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1023/B:MICS.0000006896.02938.49

Louis Saint-Amant, Pierre Drapeau

引用次数: 22

Short-term serum supplementation improves glucose-oxygen deprivation in primary cortical cultures grown under serum-free conditions. 短期补充血清改善无血清条件下生长的原代皮质培养物的葡萄糖-氧剥夺。

Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1007/s11022-004-9121-9

Jan Lewerenz, Susanne Thomsen, Julius A Steinbeck, Axel Methner

{"title":"Short-term serum supplementation improves glucose-oxygen deprivation in primary cortical cultures grown under serum-free conditions.","authors":"Jan Lewerenz, Susanne Thomsen, Julius A Steinbeck, Axel Methner","doi":"10.1007/s11022-004-9121-9","DOIUrl":"https://doi.org/10.1007/s11022-004-9121-9","url":null,"abstract":"Brain ischemia can be studied in vitro by depriving primary neurons of oxygen and glucose by replacing oxygen with argon and glucose with its antimetabolite 2-deoxy-D-glucose. In this contribution, we explain how to construct a reliably functioning ischemia chamber and use it to study neuronal cell death in neuron-enriched fetal primary cortical cultures grown under serum-free conditions. We observed that these cultures exhibited a significant cell death even during exposure to oxygenated balanced salt solution used as control for oxygen-glucose deprivation. We show that addition of only 2% fetal calf serum 24 h prior, during, and after treatment almost abolished this undesirable cell loss and proportionally increased cell death induced by oxygen-glucose deprivation. Western blots and immunocytochemistry showed that these effects were mainly due to an increase in neuronal viability under control conditions accompanied by a limited glial proliferation independent of the treatment condition. Under these modified conditions, the cultures could also still be effectively preconditioned by a short-term oxygen-glucose deprivation. In summary, this modified protocol combines the advantages of serum-free neuronal culture, where potentially toxic antimitotic substances can be omitted, with a serum-mediated protection of neurons against unspecific factors and concomitant sensitization for oxygen-glucose deprivation.","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"25 3-4","pages":"227-36"},"PeriodicalIF":0.0,"publicationDate":"2003-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s11022-004-9121-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25203831","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

Intracellular Ca2+ measurements in live cells by rapid line scan confocal microscopy: simplified calibration methodology. 细胞内Ca2+测量活细胞通过快速线扫描共聚焦显微镜:简化的校准方法。

Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1007/s11022-004-2043-8

David M Plank, Mark A Sussman

{"title":"Intracellular Ca2+ measurements in live cells by rapid line scan confocal microscopy: simplified calibration methodology.","authors":"David M Plank, Mark A Sussman","doi":"10.1007/s11022-004-2043-8","DOIUrl":"https://doi.org/10.1007/s11022-004-2043-8","url":null,"abstract":"Altered intracellular Ca2+ dynamics are characteristically observed in cardiomyocytes from failing hearts. Studies of Ca2+ handling in myocytes predominantly use Fluo-3 AM, a visible light excitable Ca2+ chelating fluorescent dye in conjunction with rapid line-scanning confocal microscopy. However, Fluo-3 AM does not allow for traditional ratiometric determination of intracellular Ca2+ concentration and has required use of mathematic correction factors with values obtained from separate procedures to convert Fluo-3 AM fluorescence to appropriate CA2+ concentrations. This study describes methodology to directly measure intracellular Ca2+ levels using inactivated, Fluo-3 AM loaded cardiomyocytes equilibrated with Ca2+ concentration standards. Titration of Ca2+ concentration exhibits a linear relationship to increasing Fluo-3 AM fluorescence intensity. Images obtained from individual myocyte confocal scans were recorded, average pixel intensity values were calculated, and a plot is generated relating the average pixel intensity to known Ca2+ concentrations. These standard plots can be used to convert transient Ca2+ fluorescence obtained with experimental cells to Ca2+ concentrations by linear regression analysis. Standards are determined on the same microscope used for acquisition of unknown Ca2+ concentrations, simplifying data interpretaton and assuring accuracy of conversion values. This procedure eliminates additional equipment, ratiometric imaging, and mathematic correction factors and should be used to investigators requiring a straightforward method for measuring Ca2+ concentrations in live cells using Ca2+-chelating dyes exhibiting variable fluorescence intensity.","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"25 3-4","pages":"123-33"},"PeriodicalIF":0.0,"publicationDate":"2003-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s11022-004-2043-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25204571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6

Establishment, characterization and viral susceptibility of 3 new cell lines from snakehead, Channa striatus (Blooch). 3个新蛇头细胞系的建立、鉴定及病毒敏感性研究。

Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1007/s11022-004-3804-0

Zhengshan Zhao, Dee Montgomery-Brock, Cheng-Sheng Lee, Yuanan Lu

{"title":"Establishment, characterization and viral susceptibility of 3 new cell lines from snakehead, Channa striatus (Blooch).","authors":"Zhengshan Zhao, Dee Montgomery-Brock, Cheng-Sheng Lee, Yuanan Lu","doi":"10.1007/s11022-004-3804-0","DOIUrl":"https://doi.org/10.1007/s11022-004-3804-0","url":null,"abstract":"Three cell lines were established from muscle (SHMS), heart (SHHT) and swim bladder (SHSB) of snakehead (Channa striatus). The cells grew initially at 25 degrees C in L15 medium supplemented with 20% fetal bovine serum and have been subcultured 13-18 times since their initiation on June 25, 2002. Growth of the snakehead cells was serum-dependent and plating efficiencies ranged from 22-29%. These snakehead cells grew well in RPMI 1640 and L-15 media, which are commonly used for cultivation of animal and mammalian cells and retained 95.9-96.6% cell viability following storage for 4 months in liquid nitrogen. Karyotyping indicated that these snakehead-derived cell lines remained diploid with a chromosome count of 44 at their early passage (passage 8-14). These cell lines were sensitive to CCV, VHSV, SVCV, IPN and SHRV; they were refractory to IHNV. These newly established cell lines are currently being used for the investigation of snakehead viral diseases in Hawaii and will be available for future isolation and study of snakehead viruses.","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"25 3-4","pages":"155-66"},"PeriodicalIF":0.0,"publicationDate":"2003-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s11022-004-3804-0","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25204575","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 29

Identifying axon guidance defects in the embryonic zebrafish brain. 鉴定胚胎斑马鱼大脑中的轴突引导缺陷。

Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1023/B:MICS.0000006851.84998.e0

Christine A Devine, Brian Key

{"title":"Identifying axon guidance defects in the embryonic zebrafish brain.","authors":"Christine A Devine, Brian Key","doi":"10.1023/B:MICS.0000006851.84998.e0","DOIUrl":"https://doi.org/10.1023/B:MICS.0000006851.84998.e0","url":null,"abstract":"The method described here outlines a simple protocol to investigate the in vivo function of axon guidance molecules during the development of the embryonic zebrafish brain. By 24 hours postfertilization, a simple scaffold of axon tracts and commissures can be visualised in the brain using acetylated alpha-tubulin, a panaxonal marker that stains all axons. The highly stereotypical trajectory of axons in the embryonic zebrafish brain provides an ideal system in which to study the molecular mechanisms of axon guidance, as defects in the axon scaffold can be clearly visualised. We describe here our approach to identify defects in the trajectory of axons that establish the initial template of tracts in the embryonic fore- and mid-brain. By combining immunohistochemical techniques and confocal microscopy on dissected wholemounts of embryonic brains we are able to observe at high resolution the complete scaffold of axon tracts. This approach provides a rapid and simple means of assessing axon guidance defects in the developing brain. Given the advantages of the zebrafish as a model system, and the range of molecular perturbation methods now available, this technique provides a valuable tool for assessing the phenotypic effects of gene perturbations in a biologically relevant context.","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"25 1-2","pages":"33-7"},"PeriodicalIF":0.0,"publicationDate":"2003-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/B:MICS.0000006851.84998.e0","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24177903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 16

Using the adult zebrafish visual system to study cadherin-2 expression during central nervous system regeneration. 利用成年斑马鱼视觉系统研究中枢神经系统再生过程中钙粘蛋白-2的表达。

Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1023/B:MICS.0000006854.18378.fc

Q Liu, R L Londraville

引用次数: 0

Model system. 模型系统。

Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01

Søren S L Andersen

引用次数: 0