Detailed Studies on the Application of Three Fluorescent Antibiotics for Dna Staining in Flow Cytometry

Crissman, Harry A.; Stevenson, Anita P.; Orlicky, David J.; Kissane, R.J.

doi:10.3109/10520297809111954

Cited by 47 publications

(16 citation statements)

References 9 publications

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“…In the majority of samples (70N, 76N, 81N, 83N, and 86N), remaining fibroblasts were largely removed at passage 2 by incubating the cultures with cold trypsin for 2-3 min. (20). One strain (70N) was karyotyped by Giemsa banding and found to be diploid with 46XX chromosomes (C.…”

Section: Establishment Of Normal Mammary Epithelial Cell Strainsmentioning

confidence: 99%

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Distinctive traits of normal and tumor-derived human mammary epithelial cells expressed in a medium that supports long-term growth of both cell types.

Band

Sager

1989

Proc. Natl. Acad. Sci. U.S.A.

232

169

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A medium is described that supports longterm growth in culture of human primary mammary tumor cells, of normal epithelial cells from mammoplasty, and of mammary tumor cell lines. Tumor cells are shown to be distinguishable from normal mammary epithelial cells by morphology, by growth requirements, and by two markers: preferential expression of the HMFG-2 epitope on tumor cells and preferential retention in tumor cell mitochondria of the lipophilic fluorescent dye rhodamine 123. Differential fluorescence of HMFG-2 fluorescein-conjugated antibodies can be used as a basis for separation of normal and tumor cells in a fluorescence-activated cell sorter, as can differential retention of rhodamine 123.

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Section: Establishment Of Normal Mammary Epithelial Cell Strainsmentioning

confidence: 99%

“…DNA content was measured as described (20 20 ,ug/ml; they were kept in the dark until fluorescence was measured at 457 nm with an Epics IV cytofluorograph.…”

mentioning

confidence: 99%

Distinctive traits of normal and tumor-derived human mammary epithelial cells expressed in a medium that supports long-term growth of both cell types.

Band

Sager

1989

Proc. Natl. Acad. Sci. U.S.A.

232

169

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“…Glutaraldehyde, unless of very good quality, may produce an autofluorescence signal in FC that can be very annoying (Booth, 1987). And formalin is known to negatively affect cell fluorescence (Crissman et al, 1978;Lebaron et al, 1998;Troussellier et al, 1999).…”

Section: Fixationmentioning

confidence: 99%

“…Crissman et al, 1978;Steen et al, 1982Steen et al, , 1986) and other stress-producing substances (e.g. Comas and Vives-Rego, 1997), and characterizing the starvation-survival response of selected bacterial species, usually those with pathogenic relevance (Thorsen et al, 1992;Lebaron and Joux, 1994).…”

Section: Bacteria and Flow Cytometrymentioning

confidence: 99%

Using flow cytometry for counting natural planktonic bacteria and understanding the structure of planktonic bacterial communities

Gasol¹,

Giorgio²

2000

Sci. Mar.

800

717

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SUMMARY: Flow cytometry is rapidly becoming a routine methodology in aquatic microbial ecology. The combination of simple to use bench-top flow cytometers and highly fluorescent nucleic acid stains allows fast and easy determination of microbe abundance in the plankton of lakes and oceans. The different dyes and protocols used to stain and count planktonic bacteria as well as the equipment in use are reviewed, with special attention to some of the problems encountered in daily routine practice such as fixation, staining and absolute counting. One of the main advantages of flow cytometry over epifluorescence microscopy is the ability to obtain cell-specific measurements in large numbers of cells with limited effort. We discuss how this characteristic has been used for differentiating photosynthetic from non-photosynthetic prokaryotes, for measuring bacterial cell size and nucleic acid content, and for estimating the relative activity and physiological state of each cell. We also describe how some of the flow cytometrically obtained data can be used to characterize the role of microbes on carbon cycling in the aquatic environment and we prospect the likely avenues of progress in the study of planktonic prokaryotes through the use of flow cytometry.

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“…Samples were allowed to equilibrate with the dye for 24 h at 4°C before analysis on an EPICS V cell sorter (Coulter Electronics Inc., Hialeah, FL). The following histograms were generated for each sample: (a) forward angle light scatter to estimate cell size (42), and (b) linear fluorescences gated from the forward angle light scatter to perform a cell cycle analysis, assess the blastogenic index (43), and measure the RNA content per cell (44 (45). Standard curves were generated using 7-amino-4-methylcoumarin (NMec; Enzyme Systems Products, Livermore, CA) in various concentrations from 0.5 to 4 MM.…”

Section: Measurement Ofprotein Degradation Ratementioning

confidence: 99%

Induction of hypertrophy in cultured proximal tubule cells by extracellular NH4Cl.

Golchini¹,

Norman²,

Bohman³

et al. 1989

J. Clin. Invest.

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Ammonia production increases in several models of renal hypertrophy in vivo. The present study was designed to determine whether ammonia can directly modulate the growth of renal cells in the absence of a change in extracellular acidity. In serum-free media NH4Cl (0-20 mM) caused JTC cells and a primary culture of rabbit proximal tubule cells to hypertrophy (increase in cell protein content) in a dose-dependent fashion without a change in DNA synthesis. Studies in JTC cells revealed that the cell protein content increased as a result of both an increase in protein synthesis and a decrease in protein degradation. Total cell RNA content and ribosome number increased after NH4Cl exposure and the cell content of the lysosomal enzymes cathepsin B and L decreased. Inhibition of the Na+/H' antiporter with amiloride did not prevent the hypertrophic response induced by NHI-Cl. The results indicate that ammonia is an important modulator of renal cell growth and that hypertrophy can occur in the absence of functioning Na+/H' antiport activity.

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Detailed Studies on the Application of Three Fluorescent Antibiotics for Dna Staining in Flow Cytometry

Cited by 47 publications

References 9 publications

Distinctive traits of normal and tumor-derived human mammary epithelial cells expressed in a medium that supports long-term growth of both cell types.

Distinctive traits of normal and tumor-derived human mammary epithelial cells expressed in a medium that supports long-term growth of both cell types.

Using flow cytometry for counting natural planktonic bacteria and understanding the structure of planktonic bacterial communities

Induction of hypertrophy in cultured proximal tubule cells by extracellular NH4Cl.

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