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Microfluidics, BioMEMS, and Medical Microsystems VII, Proc. of SPIE. 12/2009; 7207.
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IEEE Transaction on Biomedical Circuits and Systems. 04/2009; 3.
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Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI, SPIE. 12/2008; 6859.
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E.M. Haglund,
M.-M. Seale-Goldsmith,
D. Dhawan,
J. Stewart,
J. Ramos-Vara,
C.L. Cooper,
L.M. Reece,
T. Husk,
D. Bergstrom,
D. Knapp, J.F. Leary
Colloidal Quantum Dots for Biomedical Applications III, SPIE. 12/2008; 6866.
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Imaging Manipulation, and Analysis of Biomolecules, Cells, and Tissues V, SPIE. 12/2007; 6441.
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Advanced Biomed. And Clinical Diagnostic Systems III, SPIE. 12/2005; 5692:199-208.
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Advanced Biomed. And Clinical Diagnostic Systems III. 12/2005; 5692:216-223.
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SPIE. 12/2004; 5318:1-11.
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SPIE. 12/2002; 4625:1-8.
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SPIE. 12/2002; 4622:204-210.
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SPIE. 12/2001; 4255:16-27.
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SPIE. 12/2001; 4260:219-225.
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SPIE. 12/2000; 3913:36-44.
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SPIE Proceedings Systems and Technologies for Clinical Diagnostics and Drug Discovery II. 12/1999; 3603:93-101.
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SPIE. 12/1999; 3604:158-169.
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ABSTRACT: The effect of glutathione (GSH) depletion by L-buthionine-[S,R]-sulphoximine (BSO) on tumor necrosis factor-alpha (TNF-alpha)-induced adhesion molecule expression and mononuclear leukocyte adhesion to human umbilical vein endothelial cells (HUVECs) was investigated. Cells with marked depletion of cytoplasmic GSH, but with an intact pool of mitochondrial GSH, only slightly enhanced TNF-alpha-induced E-selectin and vascular cell adhesion molecule-1 (VCAM-1) expression, compared with the control. However, TNF-a-induced expression of both molecules was markedly enhanced when the mitochondrial GSH pool was diminished to <15% of the control. In contrast, TNF-alpha-induced intercellular adhesion molecule-1 (ICAM-1) expression was not affected by the depletion of either cytoplasmic or mitochondrial GSH. Marked enhancement of TNF-alpha-induced adhesion molecule expression by the depletion of mitochondrial GSH resulted in increased in mononuclear leukocyte adhesion to treated HUVECs, compared with the control. These effects parallel reactive oxygen species (ROS) formation by the depletion of mitochondrial but not cytoplasmic GSH. Our findings demonstrate that depletion of mitochondrial GSH renders more ROS generation in HUVECs, and mitochondrial GSH modulates TNF-alpha-induced adhesion molecule expression and mononuclear leukocyte adhesion in HUVECs.
Free Radical Biology and Medicine 08/1999; 27(1-2):100-9. · 5.42 Impact Factor
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ABSTRACT: As flow cytometric data becomes more complex, it becomes increasingly difficult to classify cells using conventional flow cytometry data techniques based on visual classification of the data by user-drawn regions. This paper shows some simple applications of multivariate statistical classification to classify flow cytometric data.
Discriminant Function Analysis (DFA) and Logistic Regression (LR) analysis techniques were evaluated with respect to their potential utility in the problem of detecting human breast cancer cells within normal bone marrow cells. Data sets having defined properties were employed to evaluate the potential utility of these statistical classification techniques whose performance was measured by ROC analysis.
Two extreme but reasonable situations are presented: (1) data where the separation of cells was obvious by visual inspection and (2) data where major overlaps in the values of the individual FCM parameters made intuitive classification improbable. Both DFA and LR analysis were able to classify the cells of each type with acceptable accuracy and yield.
The excellent empirical performance of both DFA and LR techniques, suggests that they offer promising approaches for classifying multiparameter FCM data using objective rules that may represent an improvement over commonly employed ad hoc approaches.
Cytometry 06/1999; 36(1):60-70.
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ABSTRACT: This paper describes new approaches to calculating the number of cells that need to be processed using flow cytometry (FCM) techniques and the subsequent time required in order to isolate a specific number of cells having selected characteristics. The methods proposed use probabilistic assumptions about the contents of the sample to be sorted, logarithmic/exponential transformations to avert the computer "underflow" and "overflow" limitations of brute force calculations for the parameters of the binomial distribution imposed by existing computer hardware, and an established mathematical procedure for calculating error bounds for the normal approximation to the binomial distribution. Estimates are derived for the total number of cells in the FCM sample volume that must be available for processing and, for given FCM cell sorting decision speeds, the total elapsed times necessary to conduct particular experiments. The proposed approach obviates the need to resort to calculation expediencies such as the theoretically limited Poisson approximation for what can be considered a Bernoulli process mathematically characterized by the binomial distribution. Tables and graphs illustrate the projected times required to complete FCM experiments as a function of "effective" cell sorting decision speeds. Results from this paper also demonstrate that, as the "effective" cell sorting decision speed increases, there may not be a corresponding linear decrease in the time required to sort a given number of cells with selected statistical properties. The focus of this paper is on the use of innovative mathematical techniques for the design of experiments involving rare cell sorting. However, these same computational approaches may also prove useful for the high-speed enrichment sorting of non-rare cell subpopulations.
Cytometry 04/1997; 27(3):233-8.
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ABSTRACT: A method to visualize the kappa opioid receptor is described that uses a high-affinity fluorescein-conjugated opioid ligand and indirect immunofluorescence with the phycoerythrin fluorophore to amplify the signal. The mouse thymoma cell line R1E/TL8x.1.G1.OUAr.1 (R1EGO), which expresses the kappa 1 but not mu or delta opioid receptors, was used as a positive control for fluorescence labeling. A fluorescein isothiocyanate-conjugated arylacetamide (FITC-AA) compound displaying high affinity for the kappa opioid receptor was synthesized. R1EGO cells were incubated with FITC-AA, in the absence or presence of the kappa-selective opioid antagonist nor-binaltorphimine (nor-BNI) as a competitor. By using fluorescence microscopy and flow cytometry, incubation of R1EGO cells with FITC-AA alone was not sufficient for the detection of specific staining of the kappa opioid receptor. To amplify the FITC-AA fluorescence, the fluorescein served as a hapten for subsequent antibody detection. R1EGO cells were incubated with FITC-AA, followed by biotinylated rabbit anti-fluorescein IgG and extravidin-conjugated R-phycoerythrin. By using this approach, R1EGO cells were stained with phycoerythrin-amplified FITC-AA, and the staining was displaced with nor-BNI. Flow cytometry showed that titrations of both FITC-AA and nor-BNI produced saturable concentration-dependent changes in the median phycoerythrin fluorescence intensity, with optimal staining at 30 microM FITC-AA. Up to 80% of the fluorescence above background was inhibited by nor-BNI. Freshly isolated thymocytes from C57BL/6ByJ mice also showed nor-BNI-sensitive staining with the FITC-AA amplification. This sensitive method of indirect phycoerythrin immunofluorescence can be used to amplify any fluorescein-conjugated opioid ligand for the detection of opioid receptors.
Proceedings of the National Academy of Sciences 03/1995; 92(4):1062-6. · 9.68 Impact Factor
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Annals of the New York Academy of Sciences 10/1994; 731:138-41. · 3.15 Impact Factor
