Fluorescent Probes of Membranes

Waggoner, Alan S.

doi:10.1007/978-1-4615-8015-7_5

Cited by 14 publications

(5 citation statements)

References 74 publications

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“…A wide range of fluorescent molecules can detect functions of living cells (for review see 20), and monoclonal antibodies conjugated with a fluorochrome have already been used in combination with FCM to quantify functional changes of mammalian spermatozoa under various experimental conditions (10). Moreover, FCM appears to allow a rapid evaluation of several parameters of cell func-tions simultaneously for each spermatozoon in a sample population (5,9).…”

Section: Discussionmentioning

confidence: 99%

Flow cytometric sorting of living, highly motile human spermatozoa based on evaluation of their mitochondrial activity.

Auger

Leonce²,

Jouannet³

et al. 1993

J Histochem Cytochem.

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We investigated the applicability of flow cytometric (FCM) sorting to select, with no deleterious effects, fractions of living, highly motile spermatozoa after staining with rhodamine 123 (Rh123) and propidium iodide (PI) for assessment of their mitochondrial activity and viability, respectively. Sperm cells were subjected to FCM sorting according to their Rh123 fluorescence intensity, and computer-aided sperm analysis (CASA) for percentage motility and movement characteristic measurements was carried out on the entire sperm populations and on the Rhl23-positive (Rh123') sorted fractions. A first experiment on five sperm samples from fertile donors pre-selected by either swim-up or simplified Percoll gradient indicated no detrimental effect of the FCM sorting procedure because: (a) the numbers of Rh123' motile sperm were not decreased by FCM sorting; (b) data on the sorted fractions showed a tendency (not significant) for an increase in movement parameters rather than a drop; and (c) a significant decrease in the percentage of PI-positive (PI+) sperm (13% vs 3%; f l 0 . 0 5 ) was measured. A second experiment was performed on sperm samples from four patients, only washed and re-suspended in Bz medium. This demonstrated a significant increase in some characteristics of movement quality related to a substantial and selective immobilization of the less motile sperm. The significant drop in the percentage of PI' sperm after FCM sorting wO.01) was less pronounced than after FCM sorting of pre-selected sperm (12% vs 3%, respectively), indicating a lethal effect of FCM sorting on a small proportion of presumably moribund sperm. These preliminary data indicate a differential effect of FCM sorting on sperm according to their function characteristics and suggest the potential importance of these methods for the characterization in vitro of sperm subpopulations on the basis of functional criteria. (J Hisrochem

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Section: Discussionmentioning

confidence: 99%

Flow cytometric sorting of living, highly motile human spermatozoa based on evaluation of their mitochondrial activity.

Auger

Leonce²,

Jouannet³

et al. 1993

J Histochem Cytochem.

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“…Cyanine dyes are a very interesting group of substances for the investigations of mitochondria (Cohen et al, 1981;Johnson et al, 1981;Laris et al, 1975;Utsumi et al, 1975;Waggoner, 1976Waggoner, , 1979. DiOCs (3,3'-dihexyloxocarbocyanine iodide) has been found to become specifically accumulated in mitochondria (Amchenkova et al, 1988;Hatano and Ueda, 1988;Korchack et al, 1982) and may stain the endoplasmic reticulum as well (Menzel and Menzel-Elsner, 1989a;Quader et al, 1987;Song et al, 1991;Weissenfels et al, 1990).…”

Section: Fluorescence Microscopymentioning

confidence: 99%

Dynamics of mitochondria in living cells: Shape changes, dislocations, fusion, and fission of mitochondria

Bereiter‐Hahn

Vöth

1994

Microscopy Res & Technique

780

496

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Mitochondria are semi-autonomous organelles which are endowed with the ability to change their shape (e.g., by elongation, shortening, branching, buckling, swelling) and their location inside a living cell. In addition they may fuse or divide. These dynamics are discussed. Dislocation of mitochondria may result from their interaction with elements of the cytoskeleton, with microtubules in particular, and from processes intrinsic to the mitochondria themselves. Morphological criteria and differences in the fate of some mitochondria argue for the presence of more than one mitochondrial population in some animal cells. Whether these reflect genetic differences remains obscure. Emphasis is laid on the methods for visualizing mitochondria in cells and following their behaviour. Fluorescence methods provide unique possibilities because of their high resolving power and because some of the mitochondria-specific fluorochromes can be used to reveal the membrane potential. Fusion and fission often occur in short time intervals within the same group of mitochondria. At sites of fusion of two mitochondria material of the inner membrane, the matrix compartment seems to accumulate. The original arrangement of the fusion partners is maintained for some minutes. Fission is a dynamic event which, like fusion, in most cases observed in vertebrate cell cultures is not a straight forward process but rather requires several "trials" until the division finally occurs. Regarding fusion and fission hitherto unpublished phase contrast micrographs, and electron micrographs have been included.

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“…Therefore, methods to estimate the ability of yeast cells to concentrate this dye were devised, using the esterase-specific dye, CF-DA. DCFH-DA and CF-DA have many similar properties and are concentrated within cells by the same esterase-dependent mechanism; however, unlike DCFH-DA, CF-DA becomes fluorescent immediately following modification by cellular esterases (30). Therefore, all cells capable of concentrating DCFH-DA display intense green fluorescence when stained with CF-DA.…”

Section: Estimation Of Dcfh-da Cell Loading Using Thementioning

confidence: 99%

Characterization of hypoxia‐dependent peroxide production in cultures of Saccharomyces cerevisiae using flow cytometry: A model for ischemic tissue destruction

Yurkow

McKenzie

1993

Cytometry

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Peroxide production in cultures of Saccharomyces cerevisiae was measured using the H202-sensitive fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) and flow cytometry. Aeration of cultures of S. cerevisiae exposed to a period of hypoxia was found to induce elevated levels of peroxide that were 100-fold higher than the levels observed in cultures maintained under exclusively aerated or hypoxic conditions. Simultaneous viability analysis, using the fluorescent DNA-intercalating dye propidium iodide, indicated that the increase in peroxide generation preceded cell damage and death. Various agents were found to influence the effect of peroxides on cell viability. The addition of ethanol to hypoxic stationary cultures dramatically increased the rate of cell death without further increasing the amount of peroxide produced, while glucose inhibited peroxide production and decreased the rate of cell death. Surprisingly, elevated peroxide levels of hypoxiclreaerated cultures were maintained upon addition of KH2P0,, although the cells remained viable for extended periods of time when compared to control and other test cultures. Similarities between our observations and those of other investigators using anoxidreperfused organs suggest that hypoxic/reaerated yeast cultures may be a useful model system to study ischemia-dependent tissue destruction of mammals.0 1993 Wiley-Liss, Inc.

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Fluorescent Probes of Membranes

Cited by 14 publications

References 74 publications

Flow cytometric sorting of living, highly motile human spermatozoa based on evaluation of their mitochondrial activity.

Flow cytometric sorting of living, highly motile human spermatozoa based on evaluation of their mitochondrial activity.

Dynamics of mitochondria in living cells: Shape changes, dislocations, fusion, and fission of mitochondria

Characterization of hypoxia‐dependent peroxide production in cultures of Saccharomyces cerevisiae using flow cytometry: A model for ischemic tissue destruction

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