Fluvo-metricell, a combined cell volume and cell fluorescence analyzer.

Kachel, Volker; Glossner, E; Kordwig, E; Ruhenstroth‐Bauer, G.

doi:10.1177/25.7.330731

Cited by 83 publications

(19 citation statements)

References 13 publications

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“…Cell volume was determined, with high accuracy, by flow cytometry by employment of an advanced Coulter system with hydrodynamic focusing (Kachel et a!., 1977;HEKA-Elektronik, Lambrecht/Pfalz, Germany). Alter ations in cell size of -1 % can be recognized with this method.…”

Section: Flow Cytometrymentioning

confidence: 99%

Swelling, Acidosis, and Irreversible Damage of Glial Cells from Exposure to Arachidonic Acid in vitro

Staub¹,

Winkler²,

Peters³

et al. 1994

J Cereb Blood Flow Metab

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Swelling and damage of C6 glioma cells and of primary cultured astrocytes were analyzed in vitro during incubation with arachidonic acid (AA; 20:4). The cells were suspended in a physiological medium supplemented with AA at concentrations of 0.001–1.0 m M. Cell swelling was quantified by flow cytometry with hydrodynamic focusing. Flow cytometry was also utilized for assessment of cell viability by exclusion of the fluorescent dye propidium iodide and for measurement of the intracellular pH (pHi) by 2′,7′-bis-(2-carboxyethyl)−5(and −6)carboxyfluorescein. Administration of AA caused an immediate dose-dependent swelling of C6 glioma cells, even at a concentration of 0.01 m M. At this level cell volume increased within 20 min to 105.0% of control, at 0.1 m M to 111.0%, while at 1.0 m M to 123.7%. Following a phase of rapid cell volume increase, swelling leveled off during the subsequent observation period of 70 min. Viability of the C6 glioma cells was 90% under control conditions. It remained unchanged after raising AA concentrations to 0.1 m M. At 0.5 m M, however, cell viability fell to 72.8%, and at 1.0 m M to 32.7%. pHi of the glioma cells was 7.3 under control conditions. In parallel with the early swelling phase, AA led to a dose-dependent decrease of the intracellular pH and an elevated lactate production of the cells. During incubation with 0.1 m M AA, pHi decreased to 7.06 after 5 min, but recovered to normal subsequently. In addition, swelling-inducing properties of linoleic (18:2) or stearic (18:0) acid were analyzed for evaluation of the specificity of glial swelling induced by AA. Whereas stearic acid (0.1 m M) failed to induce a swelling response, linoleic acid (0.1 m M) was found to be effective. The volume increase of the glial cells, however, was only half of that found during exposure to AA at the same concentration. Further, glial swelling from AA or linoleic acid was completely inhibited by the aminosteroid U-74389F, an antagonist of lipid peroxidation. Finally, omission of Na+ ions in the suspension medium with replacement by choline led also to inhibition of the cell volume increase by AA. Experiments using astrocytes from primary culture confirmed the swelling-inducing properties of AA at a quantitative level, whereas vulnerability of the cells to AA was increased. The present results demonstrate an important role of AA in cytotoxic swelling and irreversible damage of glial cells at concentrations that occur in vivo in cerebral ischemia or trauma. The damaging potential of AA might be enhanced by a concurrently evolving intracellular acidosis, stimulating the formation of oxygen-derived free radicals and lipid peroxidation.

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Section: Flow Cytometrymentioning

confidence: 99%

Swelling, Acidosis, and Irreversible Damage of Glial Cells from Exposure to Arachidonic Acid in vitro

Staub¹,

Winkler²,

Peters³

et al. 1994

J Cereb Blood Flow Metab

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“…The volume and the membrane-bound fluorescence of the stained cells were determined simultaneously with a Fluvo-Metricell flow cytometer (14), which combines electrical cell sizing and epi-illumination fluorescence measurement. The orifice for volume sizing was cylindrical, with 85 pm diameter and 100 pm length.…”

Section: Flow Cytometric Analysismentioning

confidence: 99%

Flow cytometric analysis of the binding of eleven lectins to human T‐ and B‐cells and to human T‐ and B‐cell lines

et al. 1984

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The relative surface binding of 11 lectins to human peripheral blood T‐ and B‐lymphocytes, to Molt‐4 and JM T‐cell lines, and to 6410 and NC37 B‐cell lines was determined by flow cytometry. The lectins from Lens culinaris (LCA), Ricinus communis (RCA), Arachis hypogaea (PNA), Abrus precatorius (APA), Ulex europaeus (UEA‐F), Sarothamnus scoparius (SAS‐F), Helix pomatia (HPA), Phaseolus coccineus (L‐PHA), Glycine max (SBA), and Triticum vulgare (WGA) were fluoresceinated and incubated with living, formaldehyde‐fixed, or neuraminidase‐treated cells. Except LCA, which preferentially bound to the two B‐cell lines tested in this study, none of the other lectins exhibited selective binding to the undifferentiated cells of the cell lines. The T‐cell lines and, in part, the peripheral blood T‐cells bound less WGA, APA, LCA, and L‐PHA than the B‐cell lines and the peripheral blood B‐cells. Binding of PNA was found only after neuraminidase treatment of the cells; the binding of PNA, HPA, and UEA‐F after neuraminidase treatment was higher for the T‐cells than the B‐cells from peripheral blood. No significant differences were detected between both cell types for RCA, ConA, SBA, and SAS‐F.

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“…Similarly, the percentage of cells in S phase as measured by autoradiography was the same independent of whether the cells were cultures at 33°C or 39°C for two days (data not shown). A more precise determination of the distribution of a cell population among specific phases of the cell cycle was achieved by cytofluorometric measurement of the amount of DNA per cell [22]. Such an analysis of BHK ts 422E cells grown at 33'C or 39°C revealed no significant differences in the proportion of cells in GI, S and Gz phase.…”

Section: Dna Synthesis In Bhk Ts 422e Cells During Transition From Rementioning

confidence: 99%

Ribosome Biosynthesis Is not Necessary for Initiation of DNA Replication

Grummt

Mayer

1979

European Journal of Biochemistry

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Synthesis of mature 28-S ribosomal RNA and 60-S ribosomal subunits is inhibited in baby hamster kidney (BHK) cell line ts 422E at non-permissive temperature (39 "C). This leads to a 66 decrease of total ribosomes per cell, a marked imbalance between the large and small ribosomal subunits in the cytoplasm and a decrease of cells per dish after prolonged culture at 39 "C. However, inhibition of ribosome synthesis does not affect progression of cells through the GI period of the cell division cycle, the length of the pre-replicative period, and the rate of entry of cells into S phase. In contrast to culture at non-permissive temperature, culture of BHK ts 422E cells in the presence of 0.04 pg/ml actinomycin D at 33 "C inhibits markedly the entry into S period. It is concluded that low doses of actinomycin D exert their inhibitory effect on cell growth by preventing maturation and transport of mRNA rather than by interfering with ribosome synthesis. Microfluorometric analysis revealed only slight differences in the distribution of BHK ts 422E cells in GI, S and GZ phases of the cycle either when cultured at 33°C or at 39°C. When too few ribosomes per cell are produced in BHK ts 422E cells at 39 "C, cells do not seem to be arrested reversibly at a specific point of the cell cycle but rather to die at random.The nucleolus has been considered to play a prominent role in cell proliferation. There are many reports which demonstrate that the nucleolus is markedly activated in cells stimulated to proliferate. An enhanced synthesis of rRNA in stimulated cells has been shown by an increased incorporation of radioactive precursors into nucleolar [l-51 or cytoplasmic rRNA [6-101 as well as by an increased activity of nucleolar RNA polymerase A [5, 11 -131. On the other hand, selective inhibition of nucleolar RNA synthesis by low doses of actinomycin or by lucanthone at the beginning of the GI period caused a delay of the entry of cells into the S phase [14-171. These findings were interpreted in terms of the dependence of DNA replication upon the contuined supply of ribosomal particles. Since, however, in all these studies only the biosynthesis of pre-rRNA was concerned and neither the processing of rRNA nor the assembly of mature ribosomal subunits has been studied, the question of whether growth stimulation of mammalian cells is necessarily correlated with an increased biosynthesis of ribosomes cannot yet be answered. The purpose of this work was to investigate Ahhreviations. BHK, baby hamster kidney; pre-rRNA, precursor to ribosomal RNA.to what extent, if at all, ribosome formation is a prerequisite for cells to traverse GI and enter S phase. We have used a temperature-sensitive mutant of BHK cells, ts 422E3, that is defective in the processing of 28-S rRNA at the non-permissive temperature [ 18, 191. The results suggest that the initiation of DNA replication is not related to an accumulation of cytoplasmic ribosomes during the GI period. EXPERIMENTAL PROCEDURESCell Culture

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Fluvo-metricell, a combined cell volume and cell fluorescence analyzer.

Cited by 83 publications

References 13 publications

Swelling, Acidosis, and Irreversible Damage of Glial Cells from Exposure to Arachidonic Acid in vitro

Swelling, Acidosis, and Irreversible Damage of Glial Cells from Exposure to Arachidonic Acid in vitro

Flow cytometric analysis of the binding of eleven lectins to human T‐ and B‐cells and to human T‐ and B‐cell lines

Ribosome Biosynthesis Is not Necessary for Initiation of DNA Replication

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