Single-cell analysis using capillary electrophoresis: Influence of surface support properties on cell injection into the capillary

Krylov, Sergey N.; Dovic̀hi, Norman J.

doi:10.1002/(sici)1522-2683(20000301)21:4<767::aid-elps767>3.0.co;2-e

Cited by 60 publications

(32 citation statements)

References 14 publications

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“…The small volume of the analyte needed in CE experiments has made possible the measurement of the cellular contents (native proteins [357], specific fluorescent-labelled or-induced proteins [358], metabolites), the monitoring of single cells [359,360] or bacteria communications [361]. Different CE-techniques like CGE or CIEF were developed over the last years and are currently used intensively in a similar manner to classical gel electrophoresis [362] in the fields of human genomics and proteomics [363,364].…”

Section: Discussionmentioning

confidence: 99%

Capillary electrophoresis – mass spectrometry: 15 years of developments and applications

Schmitt‐Kopplin¹,

Frommberger²

2003

Electrophoresis

282

232

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Capillary electrophoresis -mass spectrometry: 15 years of developments and applicationsSince its introduction in 1987, capillary electrophoresis-mass spectrometry (CE-MS) has developed to a well accepted multidimensional analytical approach complementary and/or competitive to classical MS-hyphenated separation techniques. The threefold combination of rapid developments of an exceptional separation technique, of selective mass detection possibilities, and of very mild ionization modes first allowed these progresses. This article shows the CE specificities that need to be well controlled/known, compared to classical and more routinely used liquid chromatography in the light of its coupling to MS. The major trends and developments over the last 15 years and most of the reviews and applications found in ISI Web of science and publisher databases are presented in a tabulated way. The reader can thus rapidly find existing CE-MS analysis techniques in his field of research and application (forensics, environment, bioanalytics, pharmaceutics, and metabolites).

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Section: Discussionmentioning

confidence: 99%

Capillary electrophoresis – mass spectrometry: 15 years of developments and applications

Schmitt‐Kopplin¹,

Frommberger²

2003

Electrophoresis

282

232

View full text Add to dashboard Cite

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“…Microscope slides were prepared by applying and spreading 5 mL of PVA stock solution (5% w/w) over each slide and allowing the slides to dry at 1407C overnight. The PVA coating on the slides prevented the cells being injected from adhering [28].…”

Section: Edta N-(2-hydroxyethyl) Piperazine-nmentioning

confidence: 99%

CE analysis of the acidic organelles of a single cell

2007

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CE analysis of the acidic organelles of a single cellThe properties of organelles within a cell have been shown to be highly heterogeneous. Until now, it has been unclear just how much of this heterogeneity is endemic to the organelle subpopulations themselves and how much is actually due to stochastic cellular noise. An attractive approach for investigating the origins of heterogeneity among the organelles of a single cell is CE with LIF detection (CE-LIF). As a proof of principle, in this report we optimize and use a single cell CE-LIF method to investigate the properties of endocytic (acidic) organelles. Our results show that the properties of individual acidic organelles containing Alexa Fluor ® 488 Dextran suggest that there are two groups of CCRF-CEM cells: a group with a high dextran content per cell, and a group with a low dextran content per cell. Furthermore, the individual organelle measurements of the single cells allow us to compare in each group the distributions of doxorubicin content per acidic organelle and electrophoretic mobilities of these organelles. Keywords IntroductionHeterogeneity among cells of the same type is known to result partly from intracellular processes, such as pinocytosis [1], the expression of proteins participating in DNA replication [2], and cellular responses to drug treatments [3], and partly from the microenvironment in which the cells reside [4]. Within the cell, heterogeneity at the subcellular level, which refers to variations in the properties of organelles of a given type, has remained practically uncharacterized up to this point. There is a great need to develop and apply single cell approaches capable of measuring the different facets of cellular heterogeneity at the subcellular level. Among the existing techniques for investigating subcellular heterogeneity, chemical imaging has been used to probe the heterogeneity of subcellular properties such as organellar pH, ion concentration, and protein interactions [5,6]. Flow cytometry [7] and CE with LIF detection (CE-LIF) [7,8] have both been used to analyze individual organelles in samples prepared from thousands to millions of cells. With all of these techniques, it has been impossible to track down which organelles originate from the same cell, information that is needed if we are to determine cell-to-cell variations.In order to determine cell-to-cell variations, single cells have been analyzed by CE [9], chromatography [10], MS [11], enzymatic radiolabeling [12], sensitive voltammetric detection [13], and flow cytometry [14]. CE has been used to describe the contents and functions of single cells (e.g., protein expression [15], release of neurotransmitters, and cell secretion [16]). In regard to the subcellular analysis of single cells, CE has been used to sample small subcellular regions, and to measure properties of the whole nucleus of a single cell [17,18]. The ability to detect individual mitochondrial particles by CE-LIF after their release from single cells has been recently described [19]. This report sho...

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“…By coating half the area of a coverslip with polyvinyl alcohol (PVA), a nonstick region was created (Krylov and Dovichi 2000). Ungerminated conidia were then trapped with the optical tweezers and carried from the PVA region to the uncoated area.…”

Section: Manipulating Whole Cells To Study Cell-cell Signallingmentioning

confidence: 99%

Experimentally manipulating fungi with optical tweezers*

et al. 2007

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A short review of the use of optical tweezers in fungal cell biological research is provided. First, we describe how optical tweezers work. Second, we review how they have been used in various experimental live-cell studies to manipulate intracellular organelles, hyphal growth and branching, and whole cells. Third, we indicate how optically trapped microbeads can be used for the localized delivery of chemicals or mechanical stimulation to cells, as well as permitting measurements of the growth forces generated by germ tubes. Finally, the effects of optical trapping on fungal cell viability and growth are assessed.

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Single-cell analysis using capillary electrophoresis: Influence of surface support properties on cell injection into the capillary

Cited by 60 publications

References 14 publications

Capillary electrophoresis – mass spectrometry: 15 years of developments and applications

Capillary electrophoresis – mass spectrometry: 15 years of developments and applications

CE analysis of the acidic organelles of a single cell

Experimentally manipulating fungi with optical tweezers*

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