Very early detection of changes associated with cellular activation using a modified flow cytometer

Kelley, Keith A.

doi:10.1002/cyto.990120512

Cited by 15 publications

(9 citation statements)

References 7 publications

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“…This can result in temporal gaps of 10 -20 s, which may preclude an accurate measurement of the initial rise in intracellular Ca 2ϩ . Several modifications to the flow cytometry equipment have been proposed to minimize the lapse in data acquisition (93,190). A relatively cheap adaptation was recently reported by Burns and Lewis (49), which allows continuous data acquisition.…”

Section: Flow Cytometrymentioning

confidence: 99%

Measurement of Intracellular Calcium

Takahashi

Camacho

Lechleiter

et al. 1999

Physiological Reviews

674

511

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To a certain extent, all cellular, physiological, and pathological phenomena that occur in cells are accompanied by ionic changes. The development of techniques allowing the measurement of such ion activities has contributed substantially to our understanding of normal and abnormal cellular function. Digital video microscopy, confocal laser scanning microscopy, and more recently multiphoton microscopy have allowed the precise spatial analysis of intracellular ion activity at the subcellular level in addition to measurement of its concentration. It is well known that Ca2+ regulates numerous physiological cellular phenomena as a second messenger as well as triggering pathological events such as cell injury and death. A number of methods have been developed to measure intracellular Ca2+. In this review, we summarize the advantages and pitfalls of a variety of Ca2+ indicators used in both optical and nonoptical techniques employed for measuring intracellular Ca2+ concentration.

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Section: Flow Cytometrymentioning

confidence: 99%

Measurement of Intracellular Calcium

Takahashi

Camacho

Lechleiter

et al. 1999

Physiological Reviews

674

511

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“…cAMP was extracted from cells with 65% ethanol. Mobilization of Ca 2+ was evaluated by the standard INDO-1 method using a modified Becton Dickinson (Mountain View, CA) FACStar Plus system as described (21). Receptor internalization.…”

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confidence: 99%

Role of the Glucagon Receptor COOH-Terminal Domain in Glucagon-Mediated Signaling and Receptor Internalization

et al. 1997

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The binding of glucagon to its hepatic receptor is known to result in a number of effects, including the intracellular accumulation of cAMP, the mobilization of intracellular Ca2+, and the endocytosis of glucagon and its receptor into intracellular vesicles. In this study, we begin to define the functional role of the COOH-terminal tail of the human glucagon receptor in glucagon-stimulated signal transduction and receptor internalization. We have created and expressed in Chinese hamster ovary (CHO) cells five truncation mutants in which the COOH-terminal 24, 56, 62, 67, and 73 amino acids have been removed. Cells expressing relevant truncated receptors were assayed for cell surface expression by immunofluorescence, for ligand-binding properties, for cAMP and Ca2+-mediated signal transduction properties, and for receptor endocytosis. In addition, a mutant receptor containing seven serine-to-alanine mutations in the COOH-terminal tail was studied. Our results reveal the following: 1) a region of the COOH-terminal tail that is required for proper cell surface expression, 2) the COOH-terminal 62 amino acids, which comprise the majority of the tail, are not required for ligand binding, cAMP accumulation, or Ca2+ mobilization, and 3) phosphorylation of the COOH-terminal tail is crucial for glucagon-stimulated receptor endocytosis.

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“…Furthermore, the demonstrated performance of ITRUs in dynamic thermoregulation is a significant improvement over previous attempts (14,15), which did not rigorously examine the actual temperature of the sample via experimental methods. Therefore, the effectiveness and accu- racy of the previous devices, as far as temperature regulation of the sample is concerned, cannot be ascertained.…”

Section: Discussionmentioning

confidence: 78%

“…Historically, sample thermoregulation in flow cytometry has been crude, relatively static, and accomplished with simple water bath devices (14,15). Water baths change temperature slowly, and thus are a poor choice for the study of dynamic temperature effects.…”

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confidence: 99%