发布求助
文献互助 智能选刊 最新文献

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

筛选
英文 中文
A rapid and reliable flow cytometric method for determining Hsp70 levels in tobacco protoplasts. 测定烟草原生质体中 Hsp70 水平的快速可靠的流式细胞计数法。
Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1007/s11022-004-2878-z
Marianne J Cronjé, Marisha Snyman, Liza Bornman, Iona E Weir
{"title":"A rapid and reliable flow cytometric method for determining Hsp70 levels in tobacco protoplasts.","authors":"Marianne J Cronjé, Marisha Snyman, Liza Bornman, Iona E Weir","doi":"10.1007/s11022-004-2878-z","DOIUrl":"10.1007/s11022-004-2878-z","url":null,"abstract":"<p><p>Current methods to determine heat shock protein (Hsp) synthesis or accumulation in plant cells, such as Western blotting and biometabolic labelling are either indirectly quantitative, labour-intensive or biohazardous. An optimal flow cytometric protocol was developed to measure the intracellular Hsp70/Hsc70 levels in tobacco protoplasts. After heat treatments, protoplasts were fixed in 2% paraformaldehyde-phosphate-buffered saline and dehydrated overnight in methyl cellusolve, followed by permeabilization with Triton X-100 (0.1% in Protoplast Wash Fluid). Immunolabelling of Hsp70/Hsc70 was done for 1 hour with a mouse monoclonal antibody and detected by R-Phycoerythrin-conjugated goat anti-mouse IgG using flow cytometry. Flow cytometry detected a significant 1.2-fold increase in Hsp70/Hsc70 accumulation (P < 0.001) in protoplasts, while Western blotting, quantified by image analysis, showed induction under similar conditions but at lower significance (P < 0.05). The coefficients of variance for flow cytometry and Western blotting were 30.7 and 49.8 respectively. Optimum temperature of heat-induced Hsp70/Hsc70 accumulation in tobacco protoplasts occurred at 40 degrees C. Flow cytometry is proposed as a quantitative, more reproducible and rapid alternative to Western blotting for the detection of Hsp70 accumulation in plant cells.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"25 3-4","pages":"237-46"},"PeriodicalIF":0.0,"publicationDate":"2003-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s11022-004-2878-z","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25203832","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: 4
A new method to obtain epithelial and stromal explants from human Corneo-Scleral Discs for the routine culture of corneal epithelial and fibroblast cells. 一种从人角膜巩膜盘获得上皮和间质外植体的新方法，用于角膜上皮和成纤维细胞的常规培养。
Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1007/s11022-004-6793-0
Sim F Webb, Sheila Davies, Richard Evans-Gowing, George Duncan
{"title":"A new method to obtain epithelial and stromal explants from human Corneo-Scleral Discs for the routine culture of corneal epithelial and fibroblast cells.","authors":"Sim F Webb,&nbsp;Sheila Davies,&nbsp;Richard Evans-Gowing,&nbsp;George Duncan","doi":"10.1007/s11022-004-6793-0","DOIUrl":"https://doi.org/10.1007/s11022-004-6793-0","url":null,"abstract":"<p><p>Acquisition of human corneal cells for culture is hindered not only by the scarcity of donor tissues but also by some of the standard enzymatic and mechanical isolation techniques. Good yields have been reported from full-thickness explant and sclero-limbal pieces. However, due to their greater proliferative capacity, fibroblasts will encroach and subsequently overwhelm epithelial cultures whichever technique is used. The novel approach presented here is to minimise this by removal of the whole stroma from the epithelial layers at the outset. This is achieved by selective sectioning with the Webb mini-microtome developed in the Norwich Eye Research Laboratory. The microtome can be sterilised by alcohol spraying or autoclaving and is small enough to use in the culture hood. A selective cut in the region of the Bowman's membrane results in the isolation of the epithelium from the stroma and thus exposed, the basal epithelial layers are released from contact inhibition to allow growth. The stroma is further cut to produce multiple sections for the culture of fibroblasts. Both pure epithelial and stromal fibroblast cultures have been successfully generated in serum-enriched medium as well as defined serum-free media with growth supplements, from the corneo-scleral discs of donors of all ages.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"25 3-4","pages":"167-76"},"PeriodicalIF":0.0,"publicationDate":"2003-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s11022-004-6793-0","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25204576","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: 2
Comparing protein stabilities during zebrafish embryogenesis. 斑马鱼胚胎发生过程中蛋白质稳定性的比较。
Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1023/B:MICS.0000006895.03556.9b
Thomas Becker, Michael Bossenz, Baris Tursun, Anne Schlüter, Marvin A Peters, Catherina G Becker, Heather P Ostendorff, Ingolf Bach
{"title":"Comparing protein stabilities during zebrafish embryogenesis.","authors":"Thomas Becker,&nbsp;Michael Bossenz,&nbsp;Baris Tursun,&nbsp;Anne Schlüter,&nbsp;Marvin A Peters,&nbsp;Catherina G Becker,&nbsp;Heather P Ostendorff,&nbsp;Ingolf Bach","doi":"10.1023/B:MICS.0000006895.03556.9b","DOIUrl":"https://doi.org/10.1023/B:MICS.0000006895.03556.9b","url":null,"abstract":"<p><p>The stabilities of many key proteins are regulated, e.g. via ubiquitination and proteasomal degradation, with important biological consequences. We present a convenient method that allows the analysis and comparison of protein stabilities during embryogenesis using early zebrafish development as a model system. Basically, this method involves ectopic overexpression of epitope-tagged proteins via mRNA injections in one-to-four-cell stage embryos and subsequent protein detection after various time points. Indeed, the protein stability of the ubiquitin ligase RLIM, which is able to autoubiquitinate and target itself for proteasomal degradation, was much shorter when compared to a protein consisting of a Myc epitope-tag and a nuclear localization domain. Thus, this method may be used more widely for the study of developmental protein stability.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"25 1-2","pages":"85-9"},"PeriodicalIF":0.0,"publicationDate":"2003-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/B:MICS.0000006895.03556.9b","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24177339","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: 4
Coulter counter use in the enumeration of muscle and fat stem cells. 库尔特计数器在肌肉和脂肪干细胞计数中的应用。
Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2003-01-01 DOI: 10.1007/978-3-642-45207-9_19
M. Fernyhough, D. Helterline, J. Vierck, R. Hill, M. Dodson
{"title":"Coulter counter use in the enumeration of muscle and fat stem cells.","authors":"M. Fernyhough, D. Helterline, J. Vierck, R. Hill, M. Dodson","doi":"10.1007/978-3-642-45207-9_19","DOIUrl":"https://doi.org/10.1007/978-3-642-45207-9_19","url":null,"abstract":"","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"9 1","pages":"221-5"},"PeriodicalIF":0.0,"publicationDate":"2003-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89351040","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
Analysis of intracellular cytokines using flowcytometry. 流式细胞术分析细胞内细胞因子。
Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2002-01-01 DOI: 10.1023/a:1024177428018
Sunil K Arora
{"title":"Analysis of intracellular cytokines using flowcytometry.","authors":"Sunil K Arora","doi":"10.1023/a:1024177428018","DOIUrl":"https://doi.org/10.1023/a:1024177428018","url":null,"abstract":"<p><p>Characterization of T-cell clones and identification of functional subsets of the helper T-cells with polarized cytokine production is based on testing of cytokine expression. Several methods have been developed that allow cytokine expression to be measured like ELISA, RT-PCR, ELISPOT, ISH and flowcytometry. Among all these methods, monitoring of cytokine production using flowcytometric analysis has its own advantages and disadvantages. Multi-parametric characterization of cytokine production on single cell basis, without long-term culture and cloning along with high throughput of samples is main feature attached to flowcytometric analysis. The interpretation may be difficult at times due to change in the phenotype of the cells. Cells with similar surface phenotype but synthesizing different cytokines and having different functional characteristics can be analyzed with this technique.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"24 1-3","pages":"37-40"},"PeriodicalIF":0.0,"publicationDate":"2002-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1024177428018","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22444200","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: 0
Nitric oxide mediated modulation of free radical generation response in the rat polymorphonuclear leukocytes: a flowcytometric study. 一氧化氮介导的大鼠多形核白细胞自由基生成反应的调节:一项流式细胞术研究。
Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2002-01-01 DOI: 10.1023/a:1024197915723
Madhu Dikshit, Prashant Sharma
{"title":"Nitric oxide mediated modulation of free radical generation response in the rat polymorphonuclear leukocytes: a flowcytometric study.","authors":"Madhu Dikshit,&nbsp;Prashant Sharma","doi":"10.1023/a:1024197915723","DOIUrl":"https://doi.org/10.1023/a:1024197915723","url":null,"abstract":"<p><p>Nitric oxide (NO) synthesis and free radical generation from polymorphonuclear leukocytes (PMNs) play an important role in several pathological conditions. It is therefore important to understand the regulatory mechanisms of free radical generation from PMNs. Flowcytometry can be used to assess generation of reactive oxygen and nitrogen species from PMNs by using fluorescent probes. In the present study regulation of NO synthesis in the control and lipopolysaccharide (LPS) treated rat PMNs has been investigated. Free radical generation was assessed by flow cytometry using a dye, 2'7'-dichlorodihydrofluorescein diacetate (DCFDA), dihydrorhodamine-123 (DHR) and 4,5-diaminofluorescein diacetate (DAF). Superoxide dismutase (SOD), and catalase significantly attenuated the arachidonic acid (AA, 1 x 10(-6) M) induced free radical generation, while 4-aminobenzoicacid hydrazide (ABH), myeloperoxidase (MPO) inhibitor had no significant effect. Intracellular and extracellular calcium levels also modulated FR generation. AA induced free radical generation from PMNs was also enhanced significantly after LPS treatment. NO synthase (NOS) inhibitors, aminoguanidine (AG) and 7-nitroindazole (NI) inhibited arachidonic acid induced free radical generation from LPS treated PMNs, while in control PMNs NOS inhibition had no effect. Augmentation of free radical generation from rat PMNs following LPS treatment seems to be regulated by NO.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"24 1-3","pages":"69-76"},"PeriodicalIF":0.0,"publicationDate":"2002-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1024197915723","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22444206","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: 0
Membrane oxidative damage and apoptosis in cervical carcinoma cells of patients after radiation therapy. 宫颈癌放疗后细胞膜氧化损伤及细胞凋亡的研究。
Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2002-01-01 DOI: 10.1023/A:1024145931652
S. Bhosle, B. Pandey, N. Huilgol, K. Mishra
{"title":"Membrane oxidative damage and apoptosis in cervical carcinoma cells of patients after radiation therapy.","authors":"S. Bhosle, B. Pandey, N. Huilgol, K. Mishra","doi":"10.1023/A:1024145931652","DOIUrl":"https://doi.org/10.1023/A:1024145931652","url":null,"abstract":"","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"22 4","pages":"65-8"},"PeriodicalIF":0.0,"publicationDate":"2002-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/A:1024145931652","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72421959","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: 17
Flowcytometric detection of PNH defect and response to therapy in aplastic anemia patients. 再生障碍性贫血患者PNH缺陷的流式细胞检测及对治疗的反应。
Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2002-01-01 DOI: 10.1007/978-94-017-0623-0_12
N. Varma, S. Varma, H. Vohra, K. Malik, G. Garewal
{"title":"Flowcytometric detection of PNH defect and response to therapy in aplastic anemia patients.","authors":"N. Varma, S. Varma, H. Vohra, K. Malik, G. Garewal","doi":"10.1007/978-94-017-0623-0_12","DOIUrl":"https://doi.org/10.1007/978-94-017-0623-0_12","url":null,"abstract":"","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"79 1","pages":"77-8"},"PeriodicalIF":0.0,"publicationDate":"2002-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90862773","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
Cell array coupled with laser scanning cytometry allows easy analysis of changes in cyclin expression during the cell cycle. An application of cell array system. 细胞阵列与激光扫描细胞术相结合，可以轻松分析细胞周期蛋白表达的变化。小区阵列系统的一个应用。
Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2002-01-01 DOI: 10.1007/978-94-017-0623-0_6
T. Furuya, Morihito Takita, S. Tsunoda, S. Kawauchi, T. Hirano, A. Oga, K. Sasaki
{"title":"Cell array coupled with laser scanning cytometry allows easy analysis of changes in cyclin expression during the cell cycle. An application of cell array system.","authors":"T. Furuya, Morihito Takita, S. Tsunoda, S. Kawauchi, T. Hirano, A. Oga, K. Sasaki","doi":"10.1007/978-94-017-0623-0_6","DOIUrl":"https://doi.org/10.1007/978-94-017-0623-0_6","url":null,"abstract":"","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"10 1","pages":"41-7"},"PeriodicalIF":0.0,"publicationDate":"2002-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85802407","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: 1
Membrane oxidative damage and apoptosis in cervical carcinoma cells of patients after radiation therapy. 宫颈癌放疗后细胞膜氧化损伤及细胞凋亡的研究。
Methods in cell science : an official journal of the Society for In Vitro Biology Pub Date : 2002-01-01
S M Bhosle, B N Pandey, N G Huilgol, K P Mishra
{"title":"Membrane oxidative damage and apoptosis in cervical carcinoma cells of patients after radiation therapy.","authors":"S M Bhosle,&nbsp;B N Pandey,&nbsp;N G Huilgol,&nbsp;K P Mishra","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>This article describes evaluation of plasma membrane fluidity and intracellular SOD with relation to apoptotic death of cervical carcinoma cells after radiation therapy. Cells from biopsies of cancer patients (stage IIIB) prior to and 24 h after radiation dose of 2 Gy were examined. Plasma membrane fluidity, measured by fluorescence polarization of DPH incorporated into lipid bilayer and superoxide dismutase (SOD) activity, determined by epinephrine method, showed significant decrease but per cent apoptotic cells, as determined by annexin-V and TUNEL methods, were found increased by two folds after radiotherapy. It is suggested that decrease in DPH polarization in membrane, reduction in SOD activity and increased apoptosis in cervical cells of cancer patients treated with radiation may be consequent to oxidative damage induced by reactive oxygen species (ROS), which may have implications in developing predictive protocol in cancer radiotherapy.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"24 1-3","pages":"65-8"},"PeriodicalIF":0.0,"publicationDate":"2002-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22444205","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: 0
首页 上一页 下一页 尾页
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整，为了账户安全，请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
请完成安全验证×
相关产品
×
本文献相关产品
联系我们：info@booksci.cn Book学术提供免费学术资源搜索服务，方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号：604180095
Book学术官方微信