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Single cell analysis using an optica...

Tufts University., Chemistry.

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Single cell analysis using an optical fiber microwell array.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Single cell analysis using an optical fiber microwell array./
Author:	Whitaker, Ragnhild Dragoey.
Description:	182 p.
Notes:	Adviser: David R. Walt.
Contained By:	Dissertation Abstracts International70-01B.
Subject:	Chemistry, Analytical. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3344060

Single cell analysis using an optical fiber microwell array.
Whitaker, Ragnhild Dragoey.

Single cell analysis using an optical fiber microwell array. - 182 p.

Adviser: David R. Walt.

Thesis (Ph.D.)--Tufts University, 2009.

Whole cells are routinely employed as biosensors in analytical research because they provide unique data about the actual cellular response to a variety of stimuli. In most conventional whole cell techniques, the signal measured is the ensemble response from thousands of cells. In the last decade, an increasing number of single cell methods have been reported, providing information that was previously unobtainable using the conventional whole cell techniques. Single cell responses are desirable because clonal populations of cells exhibit variances in observed phenotypes due to the stochastic nature of intracellular processes. Methods that detect a population average are less relevant when cells in the population deviate from this average. Single cell methods can detect differences between the cells, and therefore provide important information about population variance, receptor responses, and toxicity. In this thesis, a fiber-optic single cell microarray platform was employed to study single cell behavior in three different biochemical systems. The first system analyzed using this array was Yeast Two Hybrid (Y2H) where the statistical variance in reporter gene expression in clonal populations was studied. It was determined that protein-protein interactions and Y2H responses could be better characterized using our fiber-optic array compared to conventional Y2H analysis methods. It was also determined that single cell responses could better distinguish between real results and artifacts in the Y2H systems. Mammalian cell lines expressing human G-protein-coupled receptors (GPCRs) were also studied on the array. These cell lines were examined in order to determine clonal responses to several agonists and to determine if specific response patterns from single cells were indicative of the type of stimuli presented to the cell. The modulation of intracellular calcium in the cells was examined using a fluorescent dye and it was seen that the fiber-optic array could determine the activation power of an agonist, outliers in a population, and response patterns in single cells. Finally, bacterial communication on a single cell level was investigated using the fiber-optic array. A communicating gram negative strain and a GFP expressing sensor strain were employed on the fiber. The dual strain single cell assay enabled us to investigate bacterial communication in environments with highly defined cellular densities and configurations and to determine how bacterial communication was affected by a variety of external factors.Subjects--Topical Terms:

586156
Chemistry, Analytical.

Single cell analysis using an optical fiber microwell array.
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035 $a (UMI)AAI3344060
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100 1 $a Whitaker, Ragnhild Dragoey. $3 1058686
245 1 0 $a Single cell analysis using an optical fiber microwell array.
300 $a 182 p.
500 $a Adviser: David R. Walt.
500 $a Source: Dissertation Abstracts International, Volume: 70-01, Section: B, page: 0261.
502 $a Thesis (Ph.D.)--Tufts University, 2009.
520 $a Whole cells are routinely employed as biosensors in analytical research because they provide unique data about the actual cellular response to a variety of stimuli. In most conventional whole cell techniques, the signal measured is the ensemble response from thousands of cells. In the last decade, an increasing number of single cell methods have been reported, providing information that was previously unobtainable using the conventional whole cell techniques. Single cell responses are desirable because clonal populations of cells exhibit variances in observed phenotypes due to the stochastic nature of intracellular processes. Methods that detect a population average are less relevant when cells in the population deviate from this average. Single cell methods can detect differences between the cells, and therefore provide important information about population variance, receptor responses, and toxicity. In this thesis, a fiber-optic single cell microarray platform was employed to study single cell behavior in three different biochemical systems. The first system analyzed using this array was Yeast Two Hybrid (Y2H) where the statistical variance in reporter gene expression in clonal populations was studied. It was determined that protein-protein interactions and Y2H responses could be better characterized using our fiber-optic array compared to conventional Y2H analysis methods. It was also determined that single cell responses could better distinguish between real results and artifacts in the Y2H systems. Mammalian cell lines expressing human G-protein-coupled receptors (GPCRs) were also studied on the array. These cell lines were examined in order to determine clonal responses to several agonists and to determine if specific response patterns from single cells were indicative of the type of stimuli presented to the cell. The modulation of intracellular calcium in the cells was examined using a fluorescent dye and it was seen that the fiber-optic array could determine the activation power of an agonist, outliers in a population, and response patterns in single cells. Finally, bacterial communication on a single cell level was investigated using the fiber-optic array. A communicating gram negative strain and a GFP expressing sensor strain were employed on the fiber. The dual strain single cell assay enabled us to investigate bacterial communication in environments with highly defined cellular densities and configurations and to determine how bacterial communication was affected by a variety of external factors.
590 $a School code: 0234.
650 4 $a Chemistry, Analytical. $3 586156
650 4 $a Chemistry, Biochemistry. $3 1017722
690 $a 0486
690 $a 0487
710 2 $a Tufts University. $b Chemistry. $3 1058685
773 0 $t Dissertation Abstracts International $g 70-01B.
790 $a 0234
790 1 0 $a Kaplan, David $e committee member
790 1 0 $a Love, Christopher $e committee member
790 1 0 $a Rybak-Akimova, Elena $e committee member
790 1 0 $a Walt, David R., $e advisor
791 $a Ph.D.
792 $a 2009
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3344060