Current Protocols in Cytometry最新文献_第9页

Large Particle Sorting to Isolate Live Parasitic Nematode Eggs 大颗粒分选分离寄生线虫卵

Current Protocols in Cytometry Pub Date : 2016-04-01 DOI: 10.1002/0471142956.cy1121s76

Alfonso J. Schmidt, T. Bouchery, G. Le Gros, Kylie M. Price

引用次数: 1

Optimized MOL-PCR for Characterization of Microbial Pathogens 优化的MOL-PCR鉴定微生物病原菌

Current Protocols in Cytometry Pub Date : 2016-01-06 DOI: 10.1002/0471142956.cy1315s75

Véronique Wuyts, Nancy H.C. Roosens, Sophie Bertrand, Kathleen Marchal, Sigrid C.J. De Keersmaecker

引用次数: 0

Quantitative Flow Cytometry Measurements in Antibodies Bound per Cell Based on a CD4 Reference 定量流式细胞术测量抗体结合每个细胞基于CD4参考

Current Protocols in Cytometry Pub Date : 2016-01-06 DOI: 10.1002/0471142956.cy0129s75

Lili Wang, Heba Degheidy, Fatima Abbasi, Howard Mostowski, Gerald Marti, Steven Bauer, Robert A. Hoffman, Adolfas K. Gaigalas

{"title":"Quantitative Flow Cytometry Measurements in Antibodies Bound per Cell Based on a CD4 Reference","authors":"Lili Wang, Heba Degheidy, Fatima Abbasi, Howard Mostowski, Gerald Marti, Steven Bauer, Robert A. Hoffman, Adolfas K. Gaigalas","doi":"10.1002/0471142956.cy0129s75","DOIUrl":"10.1002/0471142956.cy0129s75","url":null,"abstract":"<p>Multicolor flow cytometer assays with fluorescently labeled antibodies are routinely used in clinical laboratories to measure the cell number of specific immunophenotypes and to estimate expression levels of specific receptors/antigens either on the cell surface or intracellularly. The cell number and specific receptors/antigens serve as biomarkers for pathological conditions at various stages of a disease. Existing methods and cell reference materials for quantitative expression measurements have not yet produced results that are of wide clinical interest or are instrument-independent across all fluorescence channels. This unit details a procedure for quantifying surface and intracellular biomarkers by calibrating the output of a multicolor flow cytometer in units of antibody bound per cell (ABC). The procedure includes (1) quality control of the flow cytometer, (2) fluorescence intensity calibration using hard dyed microspheres assigned with fluorescence intensity values, (3) compensation for fluorescence spillover between adjacent fluorescence channels, and (4) application of a biological reference calibrator to establish an ABC scale. The unit also points out current efforts for quantifying biomarkers in a manner that is independent of instrument platforms and reagent differences. © 2016 by John Wiley & Sons, Inc.</p>","PeriodicalId":11020,"journal":{"name":"Current Protocols in Cytometry","volume":"75 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/0471142956.cy0129s75","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50821487","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: 21

Simultaneous, Single-Cell Measurement of Messenger RNA, Cell Surface Proteins, and Intracellular Proteins 同时，单细胞测量信使RNA，细胞表面蛋白和细胞内蛋白

Current Protocols in Cytometry Pub Date : 2016-01-06 DOI: 10.1002/0471142956.cy0745s75

Kah Teong Soh, Joseph D. Tario Jr., Sean Colligan, Orla Maguire, Dalin Pan, Hans Minderman, Paul K. Wallace

{"title":"Simultaneous, Single-Cell Measurement of Messenger RNA, Cell Surface Proteins, and Intracellular Proteins","authors":"Kah Teong Soh, Joseph D. Tario Jr., Sean Colligan, Orla Maguire, Dalin Pan, Hans Minderman, Paul K. Wallace","doi":"10.1002/0471142956.cy0745s75","DOIUrl":"10.1002/0471142956.cy0745s75","url":null,"abstract":"<p>Nucleic acid content can be quantified by flow cytometry through the use of intercalating compounds; however, measuring the presence of specific sequences has hitherto been difficult to achieve by this methodology. The primary obstacle to detecting discrete nucleic acid sequences by flow cytometry is their low quantity and the presence of high background signals, rendering the detection of hybridized fluorescent probes challenging. Amplification of nucleic acid sequences by molecular techniques such as in situ PCR have been applied to single-cell suspensions, but these approaches have not been easily adapted to conventional flow cytometry. An alternative strategy implements a Branched DNA technique, comprising target-specific probes and sequentially hybridized amplification reagents, resulting in a theoretical 8,000- to 16,000-fold increase in fluorescence signal amplification. The Branched DNA technique allows for the quantification of native and unmanipulated mRNA content with increased signal detection and reduced background. This procedure utilizes gentle fixation steps with low hybridization temperatures, leaving the assayed cells intact to permit their concomitant immunophenotyping. This technology has the potential to advance scientific discovery by correlating potentially small quantities of mRNA with many biological measurements at the single-cell level. © 2016 by John Wiley & Sons, Inc.</p>","PeriodicalId":11020,"journal":{"name":"Current Protocols in Cytometry","volume":"75 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/0471142956.cy0745s75","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50822524","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: 14

Microscopic Investigation of Protein Function in C. elegans Using Fluorescent Imaging 利用荧光成像技术研究秀丽隐杆线虫蛋白质功能

Current Protocols in Cytometry Pub Date : 2015-10-01 DOI: 10.1002/0471142956.cy1241s74

Cliff J. Luke, Linda P. O'Reilly

引用次数: 4

In Vivo Immuno-Spin Trapping: Imaging the Footprints of Oxidative Stress 体内免疫自旋捕获:氧化应激的成像足迹

Current Protocols in Cytometry Pub Date : 2015-10-01 DOI: 10.1002/0471142956.cy1242s74

Nicholas K.H. Khoo, Nadiezhda Cantu-Medellin, Claudette St. Croix, Eric E. Kelley

{"title":"In Vivo Immuno-Spin Trapping: Imaging the Footprints of Oxidative Stress","authors":"Nicholas K.H. Khoo, Nadiezhda Cantu-Medellin, Claudette St. Croix, Eric E. Kelley","doi":"10.1002/0471142956.cy1242s74","DOIUrl":"10.1002/0471142956.cy1242s74","url":null,"abstract":"A plethora of disease processes are associated with elevated reactive species formation and allied reactions with biomolecules that alter cell signaling, induce overt damage, and promote dysfunction of tissues. Unfortunately, effective detection of reactive species in tissues is wrought with issues that significantly limit capacity for validating species identity, establishing accurate concentrations, and identifying anatomic sites of production. These shortcomings reveal the pressing need for new approaches to more precisely assess reactive species generation in vivo. Herein, we describe an in vivo immuno‐spin trapping method for indirectly assessing oxidant levels by detecting free radicals resulting from reaction of oxidants with biomolecules to form stable, immunologically detectable nitrone‐biomolecular adducts. This process couples the reactivity and sensitivity of an electron paramagnetic resonance spin trap with the resolution of confocal imaging to visualize the extent of cell and tissue oxidation and anatomic sites of production by detecting resultant free radical formation. © 2015 by John Wiley & Sons, Inc.","PeriodicalId":11020,"journal":{"name":"Current Protocols in Cytometry","volume":"74 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2015-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/0471142956.cy1242s74","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34048854","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: 13

Visualization of Telomere Integrity and Function In Vitro and In Vivo Using Immunofluorescence Techniques 利用免疫荧光技术在体外和体内可视化端粒完整性和功能

Current Protocols in Cytometry Pub Date : 2015-07-01 DOI: 10.1002/0471142956.cy1240s73

Anthony J. Cesare, Christopher M. Heaphy, Roderick J. O'Sullivan

引用次数: 42

Collection, Storage, and Preparation of Human Blood Cells 人类血细胞的收集、储存和制备

Current Protocols in Cytometry Pub Date : 2015-07-01 DOI: 10.1002/0471142956.cy0501s73

Pradeep K. Dagur, J. Philip McCoy Jr.

引用次数: 103

High Throughput-Based Mitochondrial Function Assays by Multi-Parametric Flow Cytometry 基于多参数流式细胞术的高通量线粒体功能测定

Current Protocols in Cytometry Pub Date : 2015-07-01 DOI: 10.1002/0471142956.cy0948s73

J. Paul Robinson, Nianyu Li, Padma Kumar Narayanan

引用次数: 2

Flow Cytometry of Extracellular Vesicles: Potential, Pitfalls, and Prospects 细胞外囊泡的流式细胞术:潜力、缺陷和前景

Current Protocols in Cytometry Pub Date : 2015-07-01 DOI: 10.1002/0471142956.cy1314s73

John P. Nolan

引用次数: 90