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Biomedical subjects

R Udkoff

Publications and source records attributed to R Udkoff.

5 recordsLinked to original sources
PubMed1981

Identification of mitogen responding lymphocytes by fluorescence polarization.

Measurements of the intensity and polarization distributions of fluorescein fluorescence in human lymphocyte populations show changes within minutes after exposure of the cells to the mitogens phytohemagglutinin and Concanavalin A. The distributions of polarization, which before mitogen exposure show essentially a single peak, after exposure develop a second peak at a lower value of polarization. Simultaneously, the intensity distributions show a shift to higher levels of intensity. These shifts can be modeled on the basis of a subpopulation of lymphocytes responding to mitogen. Interpretation of the results is complicated by many factors that may influence the shape of the polarization distributions. In particular we show that the measurements are sensitive to the concentrations of phytohemagglutinin, Concanavalin A, Ca++, K+, fluorescein diacetate, the incubation time and the specific donor. Nevertheless, fluorescein fluorescence measurements provide a rapid and sensitive method for studying the physiology of lymphocytes and for identifying responding lymphocyte subpopulations.

Calcium↗
PubMed1979

Polarization of fluorescein fluorescence in single cells.

Measurement of fluorescence polarization (P) gives information about the immediate environment of the fluorescent molecule. We used a flow polarimeter to investigate the factors influencing P of fluorescein in mammalian cells to determine whether such measurements are useful for characterizing heterogeneous cell populations. Fluorescein was introduced into cells by incubation with FDA. Measurements of the intensity of fluorescence (TI) and polarization (P) revealed an unexpected dependence: P decreased with increasing intensity of fluorescence. This may be accounted for by the classical model of the binding of small molecules to protein in which P is dependent on the ratio bound to unbound molecules. We have been able to estimate the quenching due to binding and construct a Scatchard plot. We estimated a wavelength shift from in vitro data consistent with the dependence of P on wavelength seen in our cell work. Generally, the distributions of P are symmetrical. Photon statistics broadens the P distribution of dim cells. However, structure does develop in the P distribution when the cells are deprived of calcium or incubated in the cold. This appears as a shoulder on the P distribution or resolves into two peaks. Calcium deprivation may differentially affect a subpopulation of cells whose significance remains to be explored in various cell types.

Animals↗
PubMed1977

Flow system fluorescence polarization measurements on fluorescein diacetate-stained EL4 cells.

We have adapted a multiparameter cell sorter to measure the distribution of fluorescence polarization in cell populations. Measurements carried out on EL4 cells show that the percent polarization of fluorescein fluorescence decreases with increasing fluorescence intensity. This inverse relationship between polarization and intensity is shown both within the cell population and by the average values of the two quantities during both the increase and decrease of fluorescence intensity. The quantitative relation between intensity and polarization is different in hypertonic than in isotonic media. These results suggest that polarization measurements carried out at a fixed time after incubation of cells with fluorescein diacetate, which is converted to fluorescein within the cells, may depend in part on the rate of fluorescein accumulation, and that agents that have been reported to change the polarization of fluorescein in living cells may do so by changing the kinetics of fluorescein accumulation.

Cell Line↗
PubMedDate not supplied

MR imaging-guided interstitial Nd:YAG laser phototherapy: dosimetry study of acute tissue damage in an in vivo model.

A dosimetry study of acute tissue damage induced by interstitial application of the neodymium-yttrium-aluminum-garnet (Nd:YAG) laser was performed with magnetic resonance (MR) imaging. The MR appearance of the lesion was correlated with gross and histopathologic findings. Seventy-six lesions were induced in rabbit muscle with laser power outputs of 5-20 W and exposure times of 20-600 seconds. MR imaging was performed immediately after laser exposure. T2-weighted spin-echo images clearly showed the acute thermal injuries caused by laser energy deposition and correlated best with histopathologic findings. These images showed three distinct layers, corresponding to central ablation, coagulative necrosis, and interstitial edema, respectively, in the pathologic findings. Lesion diameters measured on MR images showed a linear correlation with those in gross sections. Lesion volume increased not only with increasing total energy delivered but with increasing power output for a fixed total energy delivered. MR imaging is an accurate modality for dosimetry studies of laser-induced acute lesions.

Animals↗