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Measuring pH of endocytic compartmen...

The University of New Mexico.

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Measuring pH of endocytic compartments during transit of internalized IgE receptors with an innovative SERS targeted nanosensor.

紀錄類型:	書目-語言資料,印刷品 : Monograph/item
正題名/作者:	Measuring pH of endocytic compartments during transit of internalized IgE receptors with an innovative SERS targeted nanosensor./
作者:	Nowak-Lovato, Kristy L.
面頁冊數:	117 p.
附註:	Adviser: Bridget Wilson.
Contained By:	Dissertation Abstracts International70-02B.
標題:	Biophysics, Medical. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3346613
ISBN:	9781109013726

Measuring pH of endocytic compartments during transit of internalized IgE receptors with an innovative SERS targeted nanosensor.
Nowak-Lovato, Kristy L.

Measuring pH of endocytic compartments during transit of internalized IgE receptors with an innovative SERS targeted nanosensor. - 117 p.

Adviser: Bridget Wilson.

Thesis (Ph.D.)--The University of New Mexico, 2008.

The development of a novel surface-enhanced Raman Scattering (SERS) biosensor that targets and activates the Fc&egr;RI receptor mediated endocytic pathway, while measuring endosomal compartment pH over time, is demonstrated. The nanosensor consists of a 60nm Au core surrounded by a Ag coating, varying the diameter of the colloid from 70-200nm. The nanosensor is capable of targeting through the Fc&egr;RI receptor mediated endocytic pathway due to the adsorption of 2,4-&egr;-dinitrophenol-L-lysine (DNP) ligand on the nanoparticle surface. The sensor reports the pH of endosomal compartments as the receptor traffics from early endosomes to late endosomes to the secretory lysosomes (range of pH 4.5-7.5) by 4-mercaptopyridine (4-MPy) adsorbed to the nanoparticle surface. The pH-sensing ligand 4-MPy demonstrates ratiometric diagnostic frequency modes near 1600 cm-1 depending on [H+] bound or unbound to pyridine. The targeting and sensing moieties do not significantly interfere with each other's function. The sensing component of the nanosensor is calibrated with in vivo measurements of rat basophil leukemia (RBL-2H3) cells, in standard buffer solutions using the ionophore, nigericin. Nigericin serves to equilibrate the external [H+] concentration with that of the internal cellular compartments. The targeting of the nanosensor is verified with a beta-hexosaminidase assay. It is also demonstrated that the targeted nanosensor is capable of making accurate cellular pH measurements in RBL-2H3 cells, out to ninety minutes after targeted nanosensor addition. The sensor also demonstrates accuracy of measuring endocytosis in abnormal cellular states; at 25°C, or with H+ flux inhibiting drugs, amiloride and bafilomycin. Whole cell, time lapse, hyperspectral image cubes demonstrating endocytic vesicular pH during Fc&egr;RI receptor mediated endocytosis over time is demonstrated.

ISBN: 9781109013726Subjects--Topical Terms:

1017681
Biophysics, Medical.

Measuring pH of endocytic compartments during transit of internalized IgE receptors with an innovative SERS targeted nanosensor.
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