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Biomedical subjects

A Waggoner

Publications and source records attributed to A Waggoner.

16 recordsLinked to original sources
PubMed2001

Guidelines for cardiac sonographer education: recommendations of the American Society of Echocardiography Sonographer Training and Education Committee.

In 1992, the American Society of Echocardiography published a report of the Sonographer Education and Training Committee's recommendations for education of sonographers who perform echocardiographic procedures. Since the publication of the original document, there has been continual progress in technology with the development of more sophisticated diagnostic applications that allow more information to be obtained from echocardiographic procedures. These recent changes in the clinical application of echocardiography should be included in all cardiac sonographer education programs. The American Society of Echocardiography, a professional society that currently represents approximately 2500 cardiac sonographers, provides these updated guidelines.

Adult↗
PubMed1996

Multiparameter fluorescence imaging microscopy: reagents and instruments.

Fluorescence imaging microscopes and fluorescent reagents have both evolved greatly in the past 10 years. Sensitive imaging cameras developed over the last decade allow detection of fluorescence signals from even a single fluorescent tag on a protein. At the same time, numerous antibody markers for growth control proteins and DNA probes for chromosomal alterations have been discovered and these can be labeled with the new multicolor fluorescent dyes for detection by microscopy. Image analysis software automatically localizes and quantifies of as many as 5-6 of these different color fluorescent markers in the individual cells of a preparation. The results is that powerful methods for multiparameter analysis of molecular structures and dynamic processes are now accessible to the pathologist.

DNA Probes↗
PubMed1995

Aneuploidy in breast cancer: a fluorescence in situ hybridization study.

Although ploidy is associated with the development and progression of most breast cancers, the value of flow cytometric ploidy as a clinical prognostic factor remains controversial. The technique of fluorescence in situ hybridization (FISH) can be used not only to determine overall ploidy, but also to assess the over-representation or under-representation of specific chromosomes in interphase cells. This information may be of prognostic value. We studied 84 primary breast cancers and 20 metastatic tumors by FISH, using chromosome-specific fluorescent centromeric probes. Of these, 100 cases were also studied by DNA flow cytometry. The FISH studies were concordant with DNA flow cytometry with regard to distinguishing aneuploid from diploid tumors in 78% of cases. The FISH data suggested that aneuploidy arises by a process of chromosome complement doubling with subsequent chromosome loss. In tumors that exhibited evidence of more than one round of chromosome complement doubling, the selective accumulation of multiple copies of specific chromosomes or chromosome segments was common. Multiple copies of chromosomes centromeres 1, 3, and 17 were accumulated selectively in the cells of individual tumors more frequently than chromosomes centromeres 7, 11, and 16. Multiple copies of chromosomes centromeres 10 and 20 were selectively accumulated only rarely, if at all. Aneuploidy in breast cancer can be divided into distinct stages using fluorescence in situ hybridization techniques. The stages of aneuploidy provide potential landmarks in the genetic evolution of this disease with possible links to chromosome-specific evolutionary changes.

Aged↗
PubMed1992

Sensitive detection of RNAs in single cells by flow cytometry.

A rapid and sensitive fluorescent in situ hybridization method has been developed to probe RNA contents of individual cells by flow cytometry. Fixed cells in suspension were hybridized with 5' end-fluorophore-labeled oligodeoxynucleotides complementary to defined regions of the RNA of interest and analyzed by flow cytometry. With this method, we monitored combinations of histone H4 mRNA, 18S rRNA and 28S rRNA levels in synchronized HeLa S3 cells by multicolor analysis. A fluorescence signal equivalent to 1800 copies of histone H4 mRNA per cell was detected with signal-to-background ratio of 5.4. If non-specific binding of the fluorophore-labeled probe can be reduced, as few as 100 copies of mRNA of the size of H4 could be detected in individual cells by flow cytometry.

Base Sequence↗
PubMed1992

Flow cytometric analysis of lymphocyte phenotypes in bronchoalveolar lavage fluid: comparison of a two-color technique with a standard immunoperoxidase assay.

Characterization of lymphocytes in bronchoalveolar fluid has provided insight into the pathogenesis of many pulmonary diseases. Identification of lymphocyte phenotypes has become highly successful due to development of specific monoclonal antibodies and reliable methods for detecting labeled cells such as flow cytometry (FCM) and immunocytochemistry. FCM permits rapid screening of many cells, but this analysis may be confounded by heterogeneity in the size and granularity of the cells being evaluated. Such heterogeneity may lead to exclusion of cells of interest and inclusion of unwanted cells. Often peripheral blood leukocytes are used to define the gate for lung lymphocytes, but this gate may be inappropriate due to considerable variation in size and granularity of cells in bronchoalveolar lavage (BAL) fluid. Here we report an alternative method for generating a gate which employed fluorescence and side scatter signals to analyze lymphocyte subsets in BAL fluid by FCM. This gating technique avoids the pitfalls inherent in using the conventional lymphocyte gate to analyze lung cells. To validate this approach, we compared the results generated by this gate and those from the conventional forward/side light scatter gate to results derived from an immunocytochemical technique (ABC) that has been extensively employed in our laboratory to identify lymphocyte subsets in blood and lavage fluid. FCM tended to underestimate the proportions of T-cell subsets compared with ABC when the conventional gate was used. Counting only cells that stained with fluorescein-conjugated anti-CD45 antibody and that had side scatter properties of lymphocytes, however, resulted in excellent agreement between FCM and ABC. It appears that the CD45+/side scatter gate includes the vast majority of lymphocytes in BAL fluid while excluding most of the nonlymphoid cells that contaminate the conventional gate. It was this latter group of cells, and erythrocytes in particular, that led to the artificially low values for lymphocyte phenotypes in BAL fluid by FCM when the conventional lymphocyte gate was used. Although erythrocytes in BAL fluid may be eliminated by hypotonic lysis, this may also result in contamination of the conventional lymphocyte gate with nuclear debris and particulates from macrophages. Despite these advantages, the fluorescence/side scatter gate may not always be optimal for the evaluation of T lymphocytes if BAL fluid contains CD45+, nonlymphoid cells with low side light scatter. In these instances, additional antibodies such as anti-CD14 and anti-CD11 may be employed to determine the size of contaminant monocytic cells and neutrophils.(ABSTRACT TRUNCATED AT 400 WORDS)

Bronchoalveolar Lavage Fluid↗
PubMed1984

Infantile cardiac fibromatosis: correlation of antemortem and necropsy findings.

In the case presented here of a 2-month-old boy with symptoms of upper respiratory infection, the physical examination, chest X-ray film and electrocardiogram led us to suspect congestive heart failure caused by either a primary or infectious cardiomyopathy or by a structural left ventricular outflow tract obstruction. The echocardiographic findings were the first evidence to suggest the presence of an intracardiac tumor. A cardiac computerized axial tomography scan supported the findings. Operation was performed, but the tumor was unresectable. Biopsies of the mass revealed fibromatosis. The infant was discharged on diuretics, procainamide and propranolol. Approximately 3 weeks following discharge, while at rest, the child suddenly became unresponsive and could not be resuscitated. Postmortem examination verified the degree of precision that can be achieved by noninvasive techniques.

Journal Article↗
PubMed1980

Binding of merocyanine 540 to normal and leukemic erythroid cells.

Normal erythroid cells and both uninduced and induced erythroleukemia cells were stained with the leukemia-specific fluorescent probe merocyanine 540 and its analogs. The external membranes of normal intact cells bound the dye, but this general low-affinity binding was completely abolished by the addition of competing serum. In contrast, erythroleukemia cells bound the dye even in the presence of serum; binding was not affected by reversing the sign of the charge carried by MC540, but was abolished upon removal of certain hydrophobic side chains. When the erythroleukemia cells were induced to differentiate, the distribution of dye-binding regions was altered by the cell such that staining became localized to one region of the membrane. Concomitantly, conconavalin A binding sites were redistributed and became localized in the same region of the membrane as the merocyanine binding sites. Merocyanine 540 is thus shown to bind to a hematopoietic surface feature whose topological distribution is subject to cellular control during differentiation. This leukemia-specific marker may be one of several eliminated during enucleation of mammalian erythroid cells.

Animals↗
PubMed1979

Isopotentiality and an optical determination of series resistance in Limulus ventral photoreceptors.

1. Photoreceptor somas in the ventral rudimentary eye of Limulus polyphemus were impaled with three micropipettes. Two micropipettes were connected in a voltage-clamp circuit and the cells were stimulated by brief flashes. The third micropipette did not measure any significant deviations from the 'clamped' voltage during responses to the flashes, in several geometries of electrode placement, even for very bright flashes. Therefore using the described techniques there is no evidence for spatial non-uniformity of intracellular voltage in the soma of these photoreceptors. 2. A voltage-sensitive dye was used to monitor light-induced changes in membrane voltage while intracellular voltage was held clamped by a feed-back circuit. With a known series resistance connected between the bath and ground the dye recorded a light-induced change in membrane voltage. When there was no added series resistance, the light-induced change was smaller and often undetectable. From these data the naturally occurring series resistance was calculated to be less than or equal to 30 k omega. 3. From these measurements, as well as from calculations for a model spherical cell, we conclude that membrane potential can be controlled to within 2 mV using our micropipette 'point clamp' methods, for all but the brightest stimuli.

Animals↗
PubMed1976

Optical probes of membrane potential.

There are two basically different mechanisms for the fluorescence and absorption changes of merocyanine, cyanine and oxonol dyes. The permeant dyes (cyanine and oxonol dyes, with delocalized charges) work by a potential-dependent accumulation mechanism. These dyes show large (up to 80%) fluorescence and absorption changes with suspensions of cells, and the changes are complete in seconds. The impermeant dyes (merocyanine dyes, with localized charges) and the permeant dyes also show optical changes that take place in fractions of milliseconds. The rapid optical changes are relatively small (less than or equal to 5 X 10(-3)) but can often be easily detected in experiments with single cells. The rapid, nonaccumulative, optical changes result from membrane-localized dye movements. Cyanine dye-absorption changes occur because of a potential-dependent partition of dye between the membrane and the adjacent aqueous region at the high dye-concentration side of the membrane. Dimers and larger aggregates are formed in the aqueous region during the change. Merocyanine dyes may also work by the same mechanism. DiS-C3-(5) is presently the best dye for measuring membrane potentials of cells, organelles, and vesicles in suspension, but several other cyanines work nearly as well (P.J. Sims, A.S. Waggoner, C.-H. Wang, J.F. Hoffman, Biochemistry 13:3315, 1974). For each system, the ratio of dye to membrane must be varied until the optimum fluorescence change is found. A separate calibration curve must be obtained for each system. For measuring fluorescence and/or absorption changes in single cells, merocyanine 540 and diBA-C4-(5) work well but produce some photodynamic damage with high intensity illumination. A rhodanine merocyanine (WW-375) gives very large absorption changes and does not damage tissue during strong illumination. As the mechanisms of the optical changes are worked out, it should be possible to design and synthesize more sensitive, less toxic dyes that are easier to calibrate. And, as the mechanisms of the optical changes are worked out, these dyes may be useful for studying the structure and dynamics of excitable membranes.

Anilino Naphthalenesulfonates↗