Mechanistic studies of lantibiotic-induced permeabilization of phospholipid vesicles.
AuthorsDriessen AJ, van den Hooven HW, Kuiper W, van de Kamp M, Sahl HG, Konings RN, Konings WN
JournalBiochemistry
PubMed ID7849020
'Nisin is a cationic polycyclic bacteriocin secreted by some lactic acid bacteria. Nisin has previously been shown to permeabilize liposomes. The interaction of nisin was analyzed with liposomes prepared of the zwitterionic phosphatidylcholine (PC) and the anionic phosphatidylglycerol (PG). Nisin induces the release of 6-carboxyfluorescein and other small anionic fluorescent ... More
Ligand/receptor internalization: a spectroscopic analysis and a comparison of ligand binding, cellular response, and internalization by human neutrophils.
'We have compared the kinetics of the responses of neutrophils to the kinetics of ligand-receptor interaction and internalization, using as a model ligand the fluoresceinated hexapeptide N-CHO-Nle-Leu-Phe-Nle-Tyr-Lys-Fluorescein (Nle, norleucine). Cellular responses, ie, membrane depolarization, enzyme (elastase) secretion, and superoxide anion (O-2) generation, are all initiated within 10 sec of the ... More
Interaction of Mycobacterium avium-containing phagosomes with the antigen presentation pathway.
AuthorsUllrich HJ, Beatty WL, Russell DG
JournalJ Immunol
PubMed ID11086039
'Pathogenic mycobacteria infect macrophages where they replicate in phagosomes that minimize contact with late endosomal/lysosomal compartments. Loading of Ags to MHC class II molecules occurs in specialized compartments with late endosomal characteristics. This points to a sequestration of mycobacteria-containing phagosomes from the sites where Ags meet MHC class II molecules. ... More
In vitro testing of chemotherapeutic drug combinations in acute myelocytic leukaemia using the fluorometric microculture cytotoxicity assay (FMCA).
AuthorsLarsson R, Fridborg H, Kristensen J, Sundström C, Nygren P
JournalBr J Cancer
PubMed ID8494730
'The fluorometric microculture cytotoxicity assay (FMCA) was employed for analysing the effect of different chemotherapeutic drug combinations and their single constituents in 44 cases of acute myelocytic leukaemia (AML). A large heterogeneity with respect to cell kill was observed for all combinations tested, the interactions ranging from antagonistic to synergistic ... More
Fluorimetric quantification of cell death in monolayer cultures and cell suspensions.
AuthorsRuiz MC, Michelangeli F, Ludert JE, Liprandi F, del Castillo JR, Chemello ME, Benaim G, Cohen E
JournalJ Biochem Biophys Methods
PubMed ID1779095
'A fluorimetric assay using ethidium bromide (EB) was employed to quantify cell death in monolayer cell cultures (MA-104 cells) in situ and isolated cell suspensions (isolated colonic cells and Leishmania). Fluorescence of EB stained cells was measured with a photometer coupled to an inverted microscope for cell monolayers or in ... More
Hidden effects of cryopreservation on quality of human spermatozoa.
AuthorsGlander HJ, Schaller J
JournalCell Tissue Bank
PubMed ID15256959
'The effects of cryopreservation on two characteristics of human spermatozoa were investigated: the early phases of disturbed plasma membrane function and the activity of enzymes in intact spermatozoa. The membrane function was detected by means of the calcium-dependent binding of fluorescein isothiocyanate (FITC)-conjugated Annexin V to sperm plasma membranes. Annexin ... More
Flow cytometric quantitation of oxidative product formation by polymorphonuclear leukocytes during phagocytosis.
AuthorsSzejda P, Parce JW, Seeds MS, Bass DA
JournalJ Immunol
PubMed ID6491287
'Stimulation of the oxidative metabolic burst of human polymorphonuclear leukocytes (PMNL) may occur by an all-or-none trigger mechanism or by a graded response to increasing stimulation of an individual cell. If the proposed all-or-none mechanism occurred during phagocytosis, a PMNL would expend all of its metabolic potential at once, yet ... More
Continuous measurement of mitochondrial pH gradients in isolated hepatocytes by difference ratio spectroscopy.
AuthorsThomas PJ, Gaspers LD, Pharr C, Thomas JA
JournalArch Biochem Biophys
PubMed ID1898020
'The pH gradient, delta pH, present across the inner mitochondrial membrane in isolated rat hepatocytes was continuously monitored with a novel spectroscopic technique that utilizes the weak acid fluorescein. Unlike most cytosolic pH indicators, such as 2'',7''-bis(carboxyethyl)-5,(6)-carboxyfluorescein (BCECF), fluorescein freely distributes between the cytosolic and mitochondrial compartments. As is typical ... More
Cytotoxic activity of topoisomerase II inhibitors in primary cultures of tumor cells from patients with human hematologic and solid tumors.
AuthorsLarsson R, Nygren P
JournalCancer
PubMed ID7954248
'BACKGROUND. Little is known about the in vitro activity and cross-resistance patterns of topoisomerase II (topo II) inhibitors in primary cultures of tumor cells from patients with different diagnoses. METHODS. The in vitro activity of the topo II inhibitors daunorubicin, doxorubicin, idarubicin, epirubicin, amsacrine, mitoxantrone, and etoposide was determined using ... More
Prediction of individual patient response to chemotherapy by the fluorometric microculture cytotoxicity assay (FMCA) using drug specific cut-off limits and a Bayesian model.
AuthorsLarsson R, Nygren P
JournalAnticancer Res
PubMed ID8267387
'The semiautomated fluorometric microculture cytotoxicity assay (FMCA) based on the measurement of fluorescence generated from cellular hydrolysis of fluorescein diacetate (FDA) to fluorescein in microtiter plates, has been used for determination of cytotoxic drug resistance of tumor cells from patients with hematological and solid tumors. In the present study we ... More
pABC11 (also known as MOAT-C and MRP5), a member of the ABC family of proteins, has anion transporter activity but does not confer multidrug resistance when overexpressed in human embryonic kidney 293 cells.
AuthorsMcAleer MA, Breen MA, White NL, Matthews N
JournalJ Biol Chem
PubMed ID10438534
'Several members of the ABC family of proteins have been implicated in multidrug resistance associated with cancer therapies. A novel member of this gene family, designated pABC11, has been identified using degenerate polymerase chain reaction. The full-length cDNA spans 5881 base pairs and encodes an open reading frame of 1437 ... More
Functional interactions of lanthanum and phospholipase D with the abscisic acid signaling effectors VP1 and ABI1-1 in rice protoplasts.
AuthorsGampala SS, Hagenbeek D, Rock CD
JournalJ Biol Chem
PubMed ID11139577
'cis,trans-Abscisic acid (ABA) plays an important role in plant growth and development, regulation of seed maturation, germination, and adaptation to environmental stresses. Knowledge of ABA mechanisms of action and the interactions of components required for ABA signal transduction is far from complete. Using transient gene expression in rice protoplasts, we ... More
Fluorescent vital stains for complementary labelling of protoplasts from Trichoderma spp.
AuthorsHarman GE, Stasz TE
JournalStain Technol
PubMed ID2464211
'In this study several fluorescent vital stains were evaluated for their ability to provide complementary vital staining of protoplasts of Trichoderma spp. for selection of heterokaryons following protoplast fusion. Tetramethyl rhodamine isothiocyanate and fluorescein isothiocyanate were rejected because they stained only a small proportion of protoplasts. Fluorescein diacetate stained all ... More
Laboratory determination of chemotherapeutic drug resistance in tumor cells from patients with leukemia, using a fluorometric microculture cytotoxicity assay (FMCA).
AuthorsLarsson R, Kristensen J, Sandberg C, Nygren P
JournalInt J Cancer
PubMed ID1730510
'An automated fluorometric microculture cytotoxicity assay (FMCA) based on the measurement of fluorescence generated from cellular hydrolysis of fluorescein diacetate (FDA) to fluorescein was employed for chemotherapeutic-drug-sensitivity testing of tumor-cell suspensions from patients with leukemia. Fluorescence was linearly related to cell number, and reproducible measurements of drug sensitivity could be ... More
Redistribution of plasma membrane proteins by electroosmosis elicits cytosolic calcium response in tumor mast cells.
AuthorsFeder TJ, Webb WW
JournalJ Cell Physiol
PubMed ID7962106
'Activation of mast cells and basophils by binding of ligands that crosslink and micro-aggregate cell surface receptors leads to a series of responses including a phosphoinositide cascade, elevation of intracellular free calcium ([Ca2+]i), morphological changes in the cell plasma membrane, and ultimately, exocytosis of granules containing histamine and other mediators ... More
Loss of halophytism by interference with SOS1 expression.
AuthorsOh DH, Leidi E, Zhang Q, Hwang SM, Li Y, Quintero FJ, Jiang X, D'Urzo MP, Lee SY, Zhao Y, Bahk JD, Bressan RA, Yun DJ, Pardo JM, Bohnert HJ,
JournalPlant Physiol
PubMed ID19571313
'The contribution of SOS1 (for Salt Overly Sensitive 1), encoding a sodium/proton antiporter, to plant salinity tolerance was analyzed in wild-type and RNA interference (RNAi) lines of the halophytic Arabidopsis (Arabidopsis thaliana)-relative Thellungiella salsuginea. Under all conditions, SOS1 mRNA abundance was higher in Thellungiella than in Arabidopsis. Ectopic expression of ... More
Effect of glyphosate on the microbial activity of two Brazilian soils.
AuthorsAraújo AS, Monteiro RT, Abarkeli RB
JournalChemosphere
PubMed ID12757780
'Glyphosate [N-(phosphonomethyl)-glycine] is a broad-spectrum, non-selective, post-emergence herbicide that is widely used in agricultural. We studied, in vitro, changes in the microbial activity of typical Hapludult and Hapludox Brazilian soils, with and without applied glyphosate. Glyphosate was applied at a rate of 2.16 mg glyphosate kg(-1) of soil and microbial ... More
Effect of mucin and polymeric gums on uptake of fluorescein esters.
AuthorsWiedmann TS, Robinson JR
JournalJ Pharm Sci
PubMed ID3440934
'The effect of three biopolymers on the mass transport of fluorescein esters into cultured epithelial cell monolayers was determined. The method involved measurement of the rate of accumulation of fluorescein after extracellular introduction of its relatively nonfluorescent ester derivatives. Calibration of the intracellular fluorescence along with cell enumeration allowed estimation ... More
A new method to determine Giardia cyst viability: correlation of fluorescein diacetate and propidium iodide staining with animal infectivity.
AuthorsSchupp DG, Erlandsen SL
JournalAppl Environ Microbiol
PubMed ID2437857
'The viability of Giardia muris cysts was studied with the fluorogenic dyes fluorescein diacetate (FDA) and propidium iodide (PI). G. muris cysts were seen to fluoresce intensely green with FDA at an excitation wavelength of 450 to 490 nm. Cysts stained with PI fluoresced bright orange at an excitation wavelength ... More
Microfluorometric evaluation of calcein acetoxymethyl ester as a probe for P-glycoprotein-mediated resistance: effects of cyclosporin A and its nonimmunosuppressive analogue SDZ PSC 833.
AuthorsLiminga G, Nygren P, Larsson R
JournalExp Cell Res
PubMed ID7910563
'A microtiter plate-based fluorometric assay for functional measurement of 170-kDa P-glycoprotein (Pgp)-mediated transport using fluorescent calcein as a probe is described. The myeloma RPMI 8226 cell line and two of its doxorubicin-resistant Pgp-expressing sublines, dox40 (high expression) and dox6 (low expression), were used as models. Nonfluorescent calcein acetoxymethyl ester (calcein/AM) ... More
Estimation of cell survival by flow cytometric quantification of fluorescein diacetate/propidium iodide viable cell number.
AuthorsRoss DD, Joneckis CC, Ordóñez JV, Sisk AM, Wu RK, Hamburger AW Nora RE, Nora RE
JournalCancer Res
PubMed ID2736519
'We report a flow cytometric method to quantify the number of viable cells remaining in suspension culture following exposure to cytotoxic drugs. Cell viability is assessed by flow cytometric measurement of cellular fluorescence after staining with fluorescein diacetate and propidium iodide in isotonic solution. The number of viable cells per ... More
In situ assessment of cell viability.
AuthorsYang H, Acker J, Chen A, McGann L
JournalCell Transplant
PubMed ID9786064
'Cryobiological studies of tissues often require the simultaneous assessment of tissue structure and in situ cellular function. Localization of damage during cryopreservation occurs as a consequence of tissue structure and morphology and as a result of biophysical constraints imposed by diffusion and heat transfer. This study used five experimental model ... More
Analysis of early apoptotic events in individual cells by fluorescence intensity and polarization measurements.
AuthorsZurgil N, Shafran Y, Fixler D, Deutsch M
JournalBiochem Biophys Res Commun
PubMed ID11820802
'Apoptosis is a dynamic process of variable duration. The ability to continuously detect the death process occurring in single or subgroups of cells is therefore very important in identifying apoptotic cells within a complex population. The Individual Cell Scanner (ICS), a multiparametric, multilaser-based scanning static cytometer, was used in the ... More
Distinct mechanisms for aerenchyma formation in leaf sheaths of rice genotypes displaying a quiescence or escape strategy for flooding tolerance.
AuthorsParlanti S, Kudahettige NP, Lombardi L, Mensuali-Sodi A, Alpi A, Perata P, Pucciariello C,
JournalAnn Bot
PubMed ID21489969
'Rice is one of the few crops able to withstand periods of partial or even complete submergence. One of the adaptive traits of rice is the constitutive presence and further development of aerenchyma which enables oxygen to be transported to submerged organs. The development of lysigenous aerenchyma is promoted by ... More
A fluorescent staining procedure for determining the viability of mycobacterial cells.
AuthorsKvach JT, Veras JR
JournalInt J Lepr Other Mycobact Dis
PubMed ID6180992
'A fluorescent staining procedure has been developed which rapidly, accurately, and economically measures the viability of mycobacterial cells. M. smegmatis and M. phlei have served as prototype organisms to establish conditions which ensure optimal staining. The staining method incorporates the use of the fatty acid ester fluorescein diacetate (FDA) and ... More
Dynamic continuity of cytoplasmic and membrane compartments between plant cells.
AuthorsBaron-Epel O, Hernandez D, Jiang LW, Meiners S, Schindler M
JournalJ Cell Biol
PubMed ID3346323
'Fluorescence photobleaching was employed to examine the intercellular movement of fluorescein and carboxyfluorescein between contiguous soybean root cells (SB-1 cell line) growing in tissue culture. Results of these experiments demonstrated movement of these fluorescent probes between cytoplasmic (symplastic) compartments. This symplastic transport was inhibited with Ca2+ in the presence of ... More
A versatile assay for the accurate, time-resolved determination of cellular viability.
AuthorsAmano T, Hirasawa K, O'Donohue MJ, Pernolle JC, Shioi Y
JournalAnal Biochem
PubMed ID12633596
'A convenient and versatile method for the accurate, time-resolved determination of cellular viability has been developed. The conventional viability indicator fluorescein diacetate (FDA), which is converted to the fluorescent compound fluorescein in living cells, was employed as a viability probe. Fluorescence emission from cells was measured using a spectrofluorimeter equipped ... More
Positive selection of mouse B and T lymphocytes and analysis of isolated populations by flow cytometry.
AuthorsButtke TM, Mallett GS, Cuchens MA
JournalJ Immunol Methods
PubMed ID6131923
'Mouse spleen cells were separated into Ig+ and Ig- populations by positive selection using petri plates coated with rabbit anti-mouse immunoglobulin. The Ig- cells were subsequently incubated with mouse monoclonal alloantisera to Thy1.2 prior to a second positive selection. The adherent populations were characterized as B (Ig+) or T (Thy1.2+) ... More
Fluorogenic substrate detection of viable intracellular and extracellular pathogenic protozoa.
AuthorsJackson PR, Pappas MG, Hansen BD
JournalScience
PubMed ID2578226
'Viable Leishmania promastigotes and amastigotes were detected by epifluorescence microscopy with fluorescein diacetate being used to mark living parasites and the nucleic acid-binding compound ethidium bromide to stain dead cells. This procedure is superior to other assays because it is faster and detects viable intracellular as well as extracellular Leishmania. ... More
Regulatory differences in the stress response of hippocampal neurons and glial cells after heat shock.
AuthorsMarcuccilli CJ, Mathur SK, Morimoto RI, Miller RJ
JournalJ Neurosci
PubMed ID8551332
'During periods of stress, cells depend on a transient, highly conserved, and regulated response to maintain homeostasis. This "heat shock response" is mediated transcriptionally by a multigene family of heat shock factors (HSF). The presence of multiple HSF suggests that activation of a given HSF is stress-specific. Using Western blot ... More
The use of fluorescein diacetate and ethidium bromide as a viability stain for isolated islets of Langerhans.
AuthorsGray DW, Morris PJ
JournalStain Technol
PubMed ID2448922
'There is a need for a simple, rapid, sensitive method for assessing the viability of isolated islets of Langerhans. In this study the fluorescent dyes fluorescein diacetate (FDA) and ethidium bromide (EB) have been used to provide a viability assay for isolated rat islets. Discrimination of living from dead islets ... More
Polar fluorescein derivatives as improved substrate probes for flow cytoenzymological assay of cellular esterases.
AuthorsDive C, Cox H, Watson JV, Workman P
JournalMol Cell Probes
PubMed ID3173358
'Fluorescein esters are employed in assays of cell viability, membrane permeability and esterase activity. The ester most widely used, fluorescein diacetate (FDA), has the disadvantage of rapid cellular efflux of its hydrolysis product fluorescein. This is particularly problematic for flow cytoenzymology (FCE), where fluorescence is measured in individual cells allowing ... More
Isolation of highly multidrug-resistant P388 cells from drug-sensitive P388/S cells by flow cytometric cell sorting.
'To investigate the spontaneous frequency of occurrence of stable multidrug-resistant cells in a population of drug-sensitive cells, we exposed drug sensitive P388/S cells to daunorubicin (dnr) for 1 h, then used fluorescence-activated cell sorting based on intracellular dnr fluorescence to isolate cells within P388/S having different intracellular content of drug. ... More
The ES-242s, novel N-methyl-D-aspartate antagonists of microbial origin, interact with both the neurotransmitter recognition site and the ion channel domain.
AuthorsToki S, Tsukuda E, Nozawa M, Nonaka H, Yoshida M, Matsuda Y
JournalJ Biol Chem
PubMed ID1378842
'ES-242-1 approximately 5 are novel microbial bioxanthracenes which do not contain nitrogen. The ES-242s inhibited the binding of [3H]TCP and [3H]CGS19755 to the N-methyl-D-aspartate (NMDA) receptor complex. They had no effect on the binding of the specific ligands for the non-NMDA receptor. The biochemical and pharmacological properties of ES-242-1 were ... More
Rapid killing of monocytes in vitro by Candida albicans yeast cells.
AuthorsDanley DL, Polakoff J
JournalInfect Immun
PubMed ID3510173
'To study the interaction between Candida albicans blastoconidia and human phagocytes, we incubated peripheral leukocytes with fungi for 1 h at 37 degrees C and stained the cells with fluorescent vital stains ethidium bromide (EB) and fluorescein diacetate. Fungi that had been phagocytosed showed little staining; however, some leukocytes containing ... More
Fluorescent labeling of blood platelets in vivo.
AuthorsTangelder GJ, Slaaf DW, Reneman RS
JournalThromb Res
PubMed ID7167880
'In rabbits, a clear microscopic image of individual, fluorescent blood platelets flowing in a mesenteric microvessel could be obtained over the whole cross-sectional area of the vessel, following an intravenous injection of a solution containing 5-15 mg of the fluorochrome Acridine Red. Most of the circulating platelets were labeled. Activation ... More
Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates.
AuthorsPeeters E, Nelis HJ, Coenye T,
JournalJ Microbiol Methods
PubMed ID18155789
'In the present study six assays for the quantification of biofilms formed in 96-well microtiter plates were optimised and evaluated: the crystal violet (CV) assay, the Syto9 assay, the fluorescein diacetate (FDA) assay, the resazurin assay, the XTT assay and the dimethyl methylene blue (DMMB) assay. Pseudomonas aeruginosa, Burkholderia cenocepacia, ... More
Progressive incorporation of propidium iodide in cultured mouse neurons correlates with declining electrophysiological status: a fluorescence scale of membrane integrity.
AuthorsMacklis JD, Madison RD
JournalJ Neurosci Methods
PubMed ID2308380
'We describe a visual assay of neuronal electrophysiologic status for use with cultured neurons, based on the exclusion of propidium iodide (PI) by intact cellular membranes. We use this fluorescent dye, which binds to nucleic acids, at concentrations suitable for long-term exposure to neurons without toxicity. We correlate the progressive ... More
Interactions of molecular probes with living cells and tissues. Part 1. Some general mechanistic proposals, making use of a simplistic Chinese box model.
AuthorsHorobin RW, Rashid F
JournalHistochemistry
PubMed ID2358379
'A simple and generalised model-termed the simplistic Chinese box [SCB] model-for the interaction of molecular probes with living systems is described. The SCB model includes the following assumptions. That living systems may be considered as built from biologically defined boxes, e.g. whole cell, nucleus, nucleoli. That movement of molecular probes ... More
Flow cytofluorimetry of fluorescein fluorescence polarization to assay lymphocyte activation.
AuthorsDimitropoulos K, Rolland JM, Nairn RC
JournalBiochem Biophys Res Commun
PubMed ID2424436
'Change in fluorescence polarization of intracellular fluorescein measured with a specially adapted flow cytometer reliably reflected subtle biophysical changes in cells, such as those accompanying increased temperature or osmolality of the suspending medium. This system was developed to monitor changes in lymphocytes one hour after stimulation with the mitogen phytohaemagglutinin, ... More
Isolation of viable tumor cells following introduction of labelled antibody to an intracellular oncogene product using electroporation.
AuthorsBerglund DL, Starkey JR
JournalJ Immunol Methods
PubMed ID2691578
'A method for labelling the intracellular ras oncogene product, p21, with a monoclonal antibody, in B16BL6 mouse melanoma cells for subsequent flow cytometric analysis and viable cell sorting is described. Permeabilization of the cells for introduction of labelled antibody was attempted using (1) lysolecithin treatment, and (2) electroporation, a much ... More
A highly reliable, sensitive, flow cytometric/fluorometric assay for the evaluation of the anti-HIV activity of antiviral compounds in MT-4 cells.
AuthorsSchols D, Pauwels R, Vanlangendonck F, Balzarini J, De Clercq E
JournalJ Immunol Methods
PubMed ID2903196
'Infection of human T4 lymphocyte MT-4 cells with human immunodeficiency virus (HIV) results in cell death 4-5 days after infection. We have now developed a highly sensitive and rapid procedure for estimating the cytopathic effect of HIV in MT-4 cells. This method is based on fluorometric as well as flow ... More
The use of fluorescein diacetate and ethidium bromide as a stain for evaluating viability of mycobacteria.
AuthorsJarnagin JL, Luchsinger DW
JournalStain Technol
PubMed ID6160651
'A method for evaluating the viability of mycobacterial cultures using fluorescein diacetate and ethidium bromide is described. Smears from 2-4 week old cultures of mycobacteria were stained with a 0.005% Dubos albumin broth dilution of fluorescein diacetate for thirty minutes at 37 C. The smears were counterstained with a 0.005% ... More
Photolysis of caged phosphatidic acid induces flagellar excision in Chlamydomonas.
AuthorsGoedhart J, Gadella TW
JournalBiochemistry
PubMed ID15065870
'Phosphatidic (PtdOH) acid formation is recognized as an important step in numerous signaling pathways in both plants and mammals. To study the role of this lipid in signaling pathways, it is of major interest to be able to increase the amount of this lipid directly. Therefore, "caged" PtdOH was synthesized, ... More
An improved method to determine cell viability by simultaneous staining with fluorescein diacetate-propidium iodide.
AuthorsJones KH, Senft JA
JournalJ Histochem Cytochem
PubMed ID2578146
'A rapid, simultaneous double-staining procedure using fluorescein diacetate (FDA) and propidium iodide (PI) is described for use in the determination of cell viability in cell suspension. Air-dried slide preparations can be made from the cell suspensions so that an accurate estimate of the viability of the cells in the original ... More
Survival factor-insensitive generation of reactive oxygen species induced by serum deprivation in neuronal cells.
AuthorsSatoh T, Sakai N, Enokido Y, Uchiyama Y, Hatanaka H
JournalBrain Res
PubMed ID8891242
'To investigate the involvement of reactive oxygen species (ROS) in neuronal apoptosis, we performed confocal and flow cytometric analysis with a ROS-specific fluorogen, 6-carboxy-2'', 7''-dichorodihydrofluorescein diacetate, di(acetoxymethyl ester) (C-DCDHF-DA). Serum deprivation significantly increased the level of ROS in PC12 cells and rat cortical neurons. N,N''-diphenyl-p-phenylenediamine (DPPD), an antioxidant, reduced ROS ... More
Fluorescein diacetate hydrolysis for determination of accelerated degradation of thiocarbamate herbicides.
AuthorsReed JP, Krueger HR, Hall FR
JournalBull Environ Contam Toxicol
PubMed ID2597800
Fluorometric test of cell membrane integrity.
AuthorsPersidsky MD, Baillie GS
JournalCryobiology
PubMed ID560940
Membrane properties of living mammalian cells as studied by enzymatic hydrolysis of fluorogenic esters.
AuthorsRotman B, Papermaster BW
JournalProc Natl Acad Sci U S A
PubMed ID5220862
Serology for automated cytotoxicity assays. Contrast fluorescence test.
AuthorsMartel JL, Jaramillo S, Allen FH, Rubinstein P
JournalVox Sang
PubMed ID4137187
Modification of the fluorescent staining method for mycobacterial cells.
AuthorsTsukiyama F, Katoh M, Matsuo Y
JournalHiroshima J Med Sci
PubMed ID6384149
Cytotoxicity in an anchorage-independent fibroblast cell line measured by a combination of fluorescent dyes.
AuthorsKemp RB
JournalMethods Mol Biol
PubMed ID7550650
Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ.
AuthorsThomas JA, Buchsbaum RN, Zimniak A, Racker E
JournalBiochemistry
PubMed ID36128
Staining for enzymatic activity after gel electrophoresis, I.
AuthorsGabriel O, Gersten DM
JournalAnal Biochem
PubMed ID1381872
Assays of cell viability: discrimination of cells dying by apoptosis.
AuthorsDarzynkiewicz Z, Li X, Gong J
JournalMethods Cell Biol
PubMed ID7861963
Physiological assessment of bacteria using fluorochromes.
AuthorsMcFeters GA, Yu FP, Pyle BH, Stewart PS
JournalJ Microbiol Methods
PubMed ID11538412
This minireview focuses on the application of fluorogenic compounds in the detection of bacteria with particular emphasis on the assessment of physiological activity using epifluorescence microscopy. Microbiological applications of several related methods will also be reviewed. ... More
A kinetic study on the enzymatic hydrolysis of fluorescein diacetate and fluorescein-di-beta-D-galactopyranoside.
AuthorsHofmann J, Sernetz M
JournalAnal Biochem
PubMed ID6614449
The kinetics of the hydrolysis of fluoresceindiacetate and fluorescein-di-beta-D-galactopyranoside were investigated by thin-layer chromatography. The time course of the concentrations of substrate, monosubstituted intermediate, and product was simulated numerically. The mathematical model takes into account the competition of substrate and intermediate and the accumulation of the intermediate at the enzyme. ... More
Fluorescence polarization assay by flow cytometry.
AuthorsRolland JM, Dimitropoulos K, Bishop A, Hocking GR, Nairn RC
JournalJ Immunol Methods
PubMed ID3881532
Fluorescence polarization measurement on cell suspensions provides a highly sensitive means for detecting subtle changes in the cells, such as occur early after lymphocyte activation or on malignant transformation. We review here the principles of fluorescence polarization, its measurement by a commercially available flow cytometer and application of such assays ... More
Intracellular pH topography: determination by a fluorescent probe.
AuthorsSlavík J
JournalFEBS Lett
PubMed ID6852257
The distribution of pH inside the yeast Endomyces magnusii was measured at 1 micron resolution using different external pH values. In a neutral buffer the pH of the cytoplasm was 6.7-7.2 in the center, decreasing to 6.0 toward the periphery of the cell. A decrease of external pH was followed ... More
Rapid assessment of microspore and pollen development stage in wheat and maize using DAPI and membrane permeabilization.
AuthorsVergne P, Delvallee I, Dumas C
JournalStain Technol
PubMed ID3424393
The use of the DNA-specific fluorochrome DAPI has been extended to stage assessment of fresh pollen in wheat and maize. Membrane permeabilization by Triton X-100 incorporated in the staining solution allows access of the fluorochrome to nuclear DNA. At all stages of gametophytic development, the nuclei can be sharply visualized. ... More
Effects of cations, ionophores and hypoglycemic sulfonylureas on the fluorescence of fluorescein-labelled pancreatic islets.
AuthorsDeleers M, Lebrun P, Malaisse WJ
JournalRes Commun Chem Pathol Pharmacol
PubMed ID6427864
The stimulation of Ca2+ inflow into pancreatic islet cells resulting from either an increase in extracellular K+ (from 5 to 37 mM) or Ca2+ (from 1 to 20 mM) concentration or the administration of tolbutamide, glibenclamide and the ionophores A23187 or X537A resulted in an apparent fall of cellular pH ... More
A fluorescence NK assay using flow cytometry.
AuthorsMcGinnes K, Chapman G, Marks R, Penny R
JournalJ Immunol Methods
PubMed ID3944470
A flow cytometric NK assay was developed in which the K562 targets were labelled with the fluorogenic substrate, carboxyfluorescein diacetate (c'FDA). This new assay compared favourably with results obtained using the conventional 51Cr-release assay. c'FDA was not toxic to target cells and did not inhibit lysis. The assay permits the ... More
Flow cytometric analysis of viability of bull sperm cells.
AuthorsMátyus L, Szabó G, Resli I, Gáspár R, Damjanovich S
JournalActa Biochim Biophys Acad Sci Hung
PubMed ID6545628
Fluorescein diacetate and propidium iodide double fluorescence labeling of animal sperm cells in combination with 55 degrees C treatment for 15 min was used to monitor the viability and heat tolerance of sperm cells by flow cytometry. The applicability of the elaborated test to detect fertility differences has been shown ... More
Mitogen-induced changes in the fluorescence polarization of fluorescein in normal human lymphocytes: a membrane event?
AuthorsBlakeslee D
JournalJ Natl Cancer Inst
PubMed ID287826
The fluorescence polarization of human lymphocytes undergoing fluorescein fluorochromasia was measured in a system in which antifluorescein IgG was used to quench the extracellular emission of fluorescein leaked from the cells. The polarization values obtained from control cells were similar to those obtained by other investigators, as was the decrease ... More
A fluorometric assay for determining cell growth in lymphocyte proliferation and lymphokine assays.
AuthorsDotsika EN, Sanderson CJ
JournalJ Immunol Methods
PubMed ID3316407
A microplate method for assessing cell growth and viability based on the hydrolysis of fluorogenic substrates by cell esterases has been investigated. Living cells incubated with fluorescein diacetate or 4-methylumbelliferyl heptanoate generate a fluorescent product which is proportional to the number of cells. This can be used as a simple ... More
Flow cytometry with crystal violet to detect intracytoplasmic fluorescence in viable human lymphocytes. Demonstration of antibody entering living cells.
AuthorsMa JA, Chapman GV, Chen SL, Penny R, Breit SN
JournalJ Immunol Methods
PubMed ID3316393
A new method is described for the detection of intracytoplasmic fluorescence and its differentiation from surface staining of viable human lymphocytes using flow cytometry after addition of crystal violet which quenches surface but not internal fluorescence. This has then been used to study antibody penetration of viable human lymphocytes, using ... More
Calcium mobilization and exocytosis after one mechanical stretch of lung epithelial cells.
AuthorsWirtz HR, Dobbs LG
JournalScience
PubMed ID2173861
Deep inflation of the lung stimulates surfactant secretion by unknown mechanisms. The hypothesis that mechanical distension directly stimulates type II cells to secrete surfactant was tested by stretching type II cells cultured on silastic membranes. The intracellular Ca2+ concentration was measured in single cells, before and after stretching. A single ... More
Fluorogenic substrate turnover in single living cells.
AuthorsSengbusch GV, Couwenbergs C, Kühner J, Müller U
JournalHistochem J
PubMed ID60305
Intracellular diffusion properties and enzyme activities in single living cells can be analysed by means of fluorogenic substrates that diffuse into the cells where they are converted into a fluorescent product by an enzymic reaction. The reaction-kinetic analysis of this process as a system of consecutive reactions provides information on ... More
Fluorescence correlation spectroscopy in small cytosolic compartments depends critically on the diffusion model used.
AuthorsGennerich A, Schild D
JournalBiophys J
PubMed ID11106632
Fluorescence correlation spectroscopy (FCS) is a powerful technique for measuring low concentrations of fluorescent molecules and their diffusion constants. In the standard case, fluorescence fluctuations are measured in an open detection volume defined by the confocal optics. However, if FCS measurements are carried out in cellular processes that confine the ... More
DNA damage detection technique applying time-resolved fluorescence measurements.
AuthorsCosa G, Vinette AL, McLean JR, Scaiano JC
JournalAnal Chem
PubMed ID12510734
A novel DNA damage detection technique based on the characteristic fluorescence lifetimes exhibited by Pico-Green-single-stranded DNA and -double-stranded DNA complexes is employed to establish the damage produced on DNA isolated from sheep white blood cells following gamma radiation. This technique, which incorporates key concepts such as alkaline unwinding buffers and ... More
A rapid analytical technique for flow cytometric analysis of cell viability using calcofluor white M2R.
AuthorsBerglund DL, Taffs RE, Robertson NP
JournalCytometry
PubMed ID3304880
Analysis of dead versus live cells is shown to be possible using Calcoflour White M2R (CFW), a fluorescent brightener. Comparison of CFW with both propidium iodide (PI) and fluorescein diacetate (FDA) was performed on a FACS 440 dual laser flow cytometer on several populations of cultured rat and mouse cell ... More
Living with genome instability: plant responses to telomere dysfunction.
AuthorsRiha K, McKnight TD, Griffing LR, Shippen DE
JournalScience
PubMed ID11230697
Loss of telomere function in metazoans results in catastrophic damage to the genome, cell cycle arrest, and apoptosis. Here we show that the mustard weed Arabidopsis thaliana can survive up to 10 generations without telomerase. The last five generations of telomerase-deficient plants endured increasing levels of cytogenetic damage, which was ... More
Increased membrane permeability of apoptotic thymocytes: a flow cytometric study.
We have recently developed a method for the separation and quantification of viable apoptotic cells without the need for permeabilisation or fixation of the cells. The method is based on the observation that apoptotic rat thymocytes fluoresce more brightly than normal cells after a brief incubation with the DNA binding ... More
Induction of M-phase entry of prophase-blocked mouse oocytes through microinjection of okadaic acid, a specific phosphatase inhibitor.
AuthorsGavin AC, Tsukitani Y, Schorderet-Slatkine S
JournalExp Cell Res
PubMed ID1701730
We report that a specific inhibitor of types 1 and 2A phosphatases, okadaic acid (OA), induces germinal vesicle break down (GVBD) and chromosome condensation when microinjected into denuded mouse oocytes maintained in prophase block by analogs of cAMP, inhibitors of phosphodiesterase, or a tumor-promoting phorbol ester. GVBD and chromosome condensation ... More
A simple way to identify non-viable cells within living plant tissue using confocal microscopy.
AuthorsTruernit E, Haseloff J,
JournalPlant Methods
PubMed ID18573203
ABSTRACT: BACKGROUND: Plant cell death is a normal process during plant development. Mutant plants may exhibit misregulation of this process, which can lead to severe growth defects. Simple ways of visualising cell death in living plant tissues can aid the study of plant development and physiology. RESULTS: Spectral variants of ... More
Rapid analytical technique for the assessment of cell metabolic activity in marine microalgae.
AuthorsDorsey J, Yentsch CM, Mayo S, McKenna C
JournalCytometry
PubMed ID2776579
A standard method for the assessment of cell viability has been developed for marine phytoplankton using an inexpensive stain, fluorescein diacetate (FDA), at .75 microM for 10 min. A flow cytometer was used as the fluorescence detector, providing an assessment of viability for each individual particle. Cell size and chlorophyll ... More
Detection of dead cells and measurement of cell killing by flow cytometry.
AuthorsKing MA
JournalJ Immunol Methods
PubMed ID10986413
A flow cytometer can quickly perform numerous quantitative, sensitive measurements on each individual cell within a large, heterogeneous population. The modern commercially available analytical instruments, which can be found in most hospitals, pathology laboratories, and cell biology research laboratories in the industrially developed countries, can now routinely measure fluorescence simultaneously ... More
Rhodamine dyes as potential agents for photochemotherapy of cancer in human bladder carcinoma cells.
AuthorsShea CR, Chen N, Wimberly J, Hasan T
JournalCancer Res
PubMed ID2736534
The phototoxicity in vitro of rhodamine 123 and tetrabromo rhodamine 123 (TBR) was compared, in order to assess their photochemotherapeutic potential. Exposure to 514.5-nm radiation from an argon ion laser caused phototoxicity in MGH-U1 bladder carcinoma cells previously treated with either dye at 10 microM for 30 min. As assessed ... More
Cell hybridization by electrofusion on filters.
AuthorsRamos C, Bonenfant D, Teissie J
JournalAnal Biochem
PubMed ID11878799
Electric field pulses induce permeabilization and associated fusogenicity in cell membranes. Electrofusion of cells is usually performed in two steps: the first is the creation of close intercellular contacts; the second is an application of electric pulses that induces membrane fusion. Very large cell contacts can be obtained by a ... More
Sensitive, reproducible and convenient fluorometric assay for the in vitro evaluation of anti-cytomegalovirus agents.
AuthorsNeyts J, Snoeck R, Schols D, Himpens B, De Clercq E
JournalJ Virol Methods
PubMed ID1666112
Fluorescein diacetate (FDA), a non-fluorescent diacetyl fluorescein ester that becomes fluorescent upon hydrolysis by cytoplasmic esterases, permitted the easy distinction by fluorometry between non-infected and human cytomegalovirus (CMV)-infected HEL cell cultures. As a result of enhanced cytoplasmic esterase activity after CMV infection, FDA-derived fluorescence intensity was brighter for infected than ... More
Mechanisms of stimulus-evoked intracellular acidification in frog nerve fibres.
AuthorsKhodorov B, Valkina O, Turovetsky V
JournalFEBS Lett
PubMed ID8137911
Measurements of cytoplasmic pH (pHi) in frog nerve fibers (sciatic nerve and its thin bundles) were performed by using fluorescein diacetate. Earlier it had been established that veratridine (VER) treatment of the nerve greatly enhances the stimulus-evoked intracellular acidification (SEIA) which becomes irreversible after blockade of the Na+/K+ pump with ... More
Lateral phase separation of phospholipids as a basis for increased permeability of membranes towards fluorescein and other chemical species.
AuthorsHumphries GM, Lovejoy JP
JournalJ Membr Biol
PubMed ID6438340
Using mouse spleen cells, before and after treatment with glutaraldehyde or mild hyperthermia, we observe a strong correlation between permeability to fluorescein and susceptibility to staining with N epsilon-dansyl-L-lysine (irrespective of the cells' ability to exclude trypan blue). We observe the same correlation using liposomes prepared from phosphatidylcholine and varying ... More
Esterase activity, exclusion of propidium iodide, and proliferation in tumor cells exposed to anticancer agents: phenomena relevant to chemosensitivity determinations.
AuthorsPavlik EJ, Flanigan RC, van Nagell JR, Hanson MB, Donaldson ES, Keaton K, Doss B, Bartmas J, Kenady DE
JournalCancer Invest
PubMed ID4052831
Cellular esterase activity and the ability to exclude propidium iodide were examined after exposing tumor cells to anticancer agents. In general, esterase activity and the ability to exclude propidium iodide continued when cells proliferated and disappeared when proliferation was inhibited. However, with a number of preparations, drug exposure inhibited proliferation ... More
Intracellular turnover of fluorescein diacetate. Influence of membrane ionic gradients on fluorescein efflux.
AuthorsProsperi E
JournalHistochem J
PubMed ID2387757
The influence of the membrane ionic gradient on the efflux of Fluorescein after intracellular turnover of Fluorescein diacetate was studied in HeLa cells. The kinetics of Fluorescein efflux was monitored by determining with flow cytometry the decrease in fluorescence intensity of single cells. Alterations of the Na+ and K+ gradients ... More
Enzyme cytochemical techniques for metabolic mapping in living cells, with special reference to proteolysis.
AuthorsBoonacker E, Van Noorden CJ
JournalJ Histochem Cytochem
PubMed ID11724895
Specific enzymes play key roles in many pathophysiological processes and therefore are targets for therapeutic strategies. The activity of most enzymes is largely determined by many factors at the post-translational level. Therefore, it is essential to study the activity of target enzymes in living cells and tissues in a quantitative ... More
Assessment of activated sludge viability with flow cytometry.
AuthorsZiglio G, Andreottola G, Barbesti S, Boschetti G, Bruni L, Foladori P, Villa R
JournalWater Res
PubMed ID11827352
The aim of the study was to evaluate the applicability of fluorescent dyes and multiparameter flow cytometry for the rapid and direct viability/activity assessment of activated sludge samples taken from wastewater treatment plants. Viability and activity of the biomass were estimated respectively through cellular membrane integrity, staining with SYBR Green ... More
Laser-micropipet combination for single-cell analysis.
Due to its potential for exquisite mass detection limits and resolving power, capillary electrophoresis is used for biochemical measurements on single cells; however, accurate measurements of many physiological parameters require sampling strategies that are considerably faster than those presently available. We have developed a laser-based technique to lyse single, adherent, ... More
Transcellular transport of fluorescein in hepatocyte monolayers: evidence for functional polarity of cells in culture.
AuthorsBarth CA, Schwarz LR
JournalProc Natl Acad Sci U S A
PubMed ID6956908
The rat liver in vivo transfers bile salts, proteins, and dyes from blood into bile. It is the purpose of this communication to demonstrate the maintenance of this transcellular transport in cultured adult rat hepatocytes. Two minutes after adding fluorescein (20 microgram/ml) to the culture medium, maximal cellular fluorescence was ... More
A rapid cell membrane permeability test using fluorescent dyes and flow cytometry.
AuthorsAeschbacher M, Reinhardt CA, Zbinden G
JournalCell Biol Toxicol
PubMed ID3267449
A reliable and rapid test to detect cytotoxic chemicals which affect cell membranes is described. Fluorescein diacetate freely penetrates intact cells where it is hydrolyzed to its fluorochrome, fluorescein, which is retained in the cell due to its polarity. On the other hand, ethidium bromide is known to be excluded ... More
Regional heterogeneity in intracellular distribution of superoxide and hydrogen peroxide within the sperm and its relation to sperm development.
AuthorsAgnihotri S, Purohit SB, Laloraya M, Kumar GP
JournalArch Androl
PubMed ID10543573
This study was performed to understand the regional distribution of superoxide anion radicals and hydrogen peroxide within the spermatozoa of mice during both normal and altered situations of epididymal maturation. The intracellular O2*- levels were probed employing dichlorodihydrofluorescein diacetate (DDF) as a reporter. The testicular spermatozoa from normal animals showed ... More
Inorganic Pi increases neuronal survival in the acute early phase following excitotoxic/oxidative insults.
AuthorsGlinn M, Ni B, Irwin RP, Kelley SW, Lin SZ, Paul SM
JournalJ Neurochem
PubMed ID9572268
Inorganic phosphate (Pi) plays a vital role in intracellular energy metabolism. Its many effects include stimulation of glucose use, enhancement of high-energy phosphate concentrations, and modulation of cytosolic free [Ca2+]. Cultured fetal rat cortical neurons constitutively import Pi, and cytosolic levels positively correlate with [ATP], [NADPH], and energy charge. In ... More
Change of intracellular fluidity during keratinocyte differentiation measured by fluorescence polarization.
AuthorsHachisuka H, Nomura H, Sasai Y, Shiotsuki K, Yokoyama MM
JournalCell Tissue Res
PubMed ID2431784
The fluorescence polarization method was applied to measure the intracellular fluidity of fractionated guinea pig keratinocytes. Guinea pig epidermal cell suspension was obtained by treatment with EDTA and trypsin, and was separated into high, intermediate, and low density fractions using Percoll density gradient centrifugation. Morphological observation and cytofluorometric analysis of ... More
Production of reactive oxygen species and release of L-glutamate during superoxide anion-induced cell death of cerebellar granule neurons.
AuthorsSatoh T, Numakawa T, Abiru Y, Yamagata T, Ishikawa Y, Enokido Y, Hatanaka H
JournalJ Neurochem
PubMed ID9422377
Enhanced production of superoxide anion (O2-) is considered to play a pivotal role in the pathogenesis of CNS neurons. Here, we report that O2- generated by xanthine (XA) + xanthine oxidase (XO) triggered cell death associated with nuclear condensation and DNA fragmentation in cerebellar granule neuron. XA + XO induced ... More
Flow cytometric analysis of membrane permeability properties influencing intracellular accumulation and efflux of fluorescein.
AuthorsProsperi E, Croce AC, Bottiroli G, Supino R
JournalCytometry
PubMed ID3948603
A flow cytometric investigation has been made on the membrane permeability properties that mediate intracellular turnover of fluorogenic substrates. The accumulation and efflux of fluorescein, consequent to the enzymatic turnover of fluorescein diacetate, were assessed in the presence of metabolic inhibitors and after treatment with membrane-active compounds. The metabolic poisons ... More
Detection of tumor-specific cytotoxic drug activity in vitro using the fluorometric microculture cytotoxicity assay and primary cultures of tumor cells from patients.
AuthorsNygren P, Fridborg H, Csoka K, Sundström C, de la Torre M, Kristensen J, Bergh J, Hagberg H, Glimelius B, Rastad J
JournalInt J Cancer
PubMed ID8314348
The semi-automated fluorometric microculture cytotoxicity assay (FMCA), based on the measurement of fluorescence generated from cellular hydrolysis of fluorescein diacetate (FDA) by viable cells, was employed for cytotoxic drug sensitivity testing of tumor cells from patients with hematological or solid tumors. In total, 390 samples from 20 diagnoses were tested ... More
Fluorescent dyes for lymphocyte migration and proliferation studies.
AuthorsParish CR
JournalImmunol Cell Biol
PubMed ID10571670
Fluorescent dyes are increasingly being exploited to track lymphocyte migration and proliferation. The present paper reviews the properties and performance of some 14 different fluorescent dyes that have been used during the last 20 years to monitor lymphocyte migration. Of the 14 dyes discussed, two stand out as being the ... More
New fluorescent dyes for lymphocyte migration studies. Analysis by flow cytometry and fluorescence microscopy.
AuthorsWeston SA, Parish CR
JournalJ Immunol Methods
PubMed ID2212694
16 fluorochromes were examined for their ability to label viable lymphocytes in vitro and yield fluorescence detectable by fluorescence microscopy and flow cytometry. Of these fluorochromes, four intracellular dyes were found to be suitable for in vivo migration studies. They were H33342, the well known DNA-binding dye which excites and ... More
Activity of cyclosporins as resistance modifiers in primary cultures of human haematological and solid tumours.
The semiautomated fluorimetric microculture cytotoxicity assay (FMCA) was used for evaluation of the ability of cyclosporin A (CsA) and its novel non-immunosuppressive derivative SDZ PSC 833 (PSC) to modify the response to doxorubicin or vincristine in vitro in different haematological and solid human tumour types. Primary cultures of 322 tumour ... More
Analysis of enzyme kinetics in individual living cells utilizing fluorescence intensity and polarization measurements.
AuthorsDeutsch M, Kaufman M, Shapiro H, Zurgil N
JournalCytometry
PubMed ID10655561
BACKGROUND: The Cellscan mark-S (CS-S) scanning cytometer was used for tracing enzymatic reactions in the same individual cells under various physiological conditions over periods of minutes. On-line reagent addition and changes in the experimental conditions (buffers, ions, substrates and inhibitors) were performed. METHODS: Kinetic events were monitored by fluorescence intensity ... More
The use of fluorochromes in the cytochemical characterization of some phytoflagellates.
AuthorsKlut ME, Bisalputra T, Antia NJ
JournalHistochem J
PubMed ID2453490
Sixteen fluorochromes were tested for the cytochemical characterization of two dinoflagellates (Amphidinuim carterae, Prorocentrum micans) and one chlorophycean flagellate (Dunaliella tertiolecta). Depending on the fluorochrome used, various cellular components (including the plasma membrane, thecal plates, pusule, trichocysts, nucleus, lipid bodies and vacuoles) were revealed. The different colours obtained from single ... More