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You searched for subject:(Fluorescence ratiometric analysis). Showing records 1 – 3 of 3 total matches.

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University of Arizona

1. Lin, Yuxiang. DEVELOPMENT AND ANALYSIS OF OPTICAL PH IMAGING TECHNIQUES .

Degree: 2010, University of Arizona

The pH of tumors and surrounding tissues is a key biophysical property of the tumor microenvironment that affects how a tumor survives and how it invades the surrounding space of normal tissue. Research into tumorigenesis and tumor treatment is greatly dependent on accurate, precise, and reproducible measurements. Optical imaging is generally regarded as the best choice for non-invasive and high spatial resolution measurements. Ratiometric fluorescence imaging and fluorescence lifetime imaging microscopy (FLIM) are two primary ways for measuring tumor pH.pH measurements in a window chamber animal model using a ratiometric fluorescence imaging technique is demonstrated in this dissertation. The experimental setup, imaging protocols, and results are presented. A significantly varying bias was consistently observed in the measured pH. A comprehensive analysis on the possible error sources accounting for this bias is carried out. The result of analysis reveals that accuracy of ratiometric method is most likely limited by biological and physiological factors.FLIM is a promising alternative because the fluorescence lifetime is insensitive to the biological and physiological factors. Photon noise is the predominant error source of FLIM. The Fisher information matrix and the Cramér-Rao lower bound are used to calculate the lowest possible variance of estimated lifetime for time-domain (TD) FLIM. A statistical analysis of frequency-domain (FD) FLIM using homodyne lock-in detection is also performed and the probability density function of the estimated lifetime is derived. The results allow the derivation of the optimum experimental parameters, which yields the lowest variance of the estimated lifetime in a given period of imaging time. The analyses of both TD and FD-FLIM agree with results of corresponding Monte Carlo simulations. Advisors/Committee Members: Gmitro, Arthur F (advisor), Utzinger, Urs (committeemember).

Subjects/Keywords: accuracy and precision; error analysis; fluorescence lifetime; optical pH imaging; ratiometric imaging; statistical analysis

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Lin, Y. (2010). DEVELOPMENT AND ANALYSIS OF OPTICAL PH IMAGING TECHNIQUES . (Doctoral Dissertation). University of Arizona. Retrieved from http://hdl.handle.net/10150/193846

Chicago Manual of Style (16th Edition):

Lin, Yuxiang. “DEVELOPMENT AND ANALYSIS OF OPTICAL PH IMAGING TECHNIQUES .” 2010. Doctoral Dissertation, University of Arizona. Accessed September 22, 2017. http://hdl.handle.net/10150/193846.

MLA Handbook (7th Edition):

Lin, Yuxiang. “DEVELOPMENT AND ANALYSIS OF OPTICAL PH IMAGING TECHNIQUES .” 2010. Web. 22 Sep 2017.

Vancouver:

Lin Y. DEVELOPMENT AND ANALYSIS OF OPTICAL PH IMAGING TECHNIQUES . [Internet] [Doctoral dissertation]. University of Arizona; 2010. [cited 2017 Sep 22]. Available from: http://hdl.handle.net/10150/193846.

Council of Science Editors:

Lin Y. DEVELOPMENT AND ANALYSIS OF OPTICAL PH IMAGING TECHNIQUES . [Doctoral Dissertation]. University of Arizona; 2010. Available from: http://hdl.handle.net/10150/193846


NSYSU

2. LIN, DING-JYUN. Heptamethine Cyanine Based Semiconducting Polymer Dots for : ( i ) Detection of Cellular pH ; and ( ii ) Quantification of Glutathione.

Degree: Master, Chemistry, 2017, NSYSU

In this study, we used the advantages of good light stability, broad absorption band, high energy transfer and low biotoxicity, easy surface modification, based fluorescent semiconductor polymer nanoparticles (Pdots) to design the detection probe. Using the FRET (Förster resonance energy transfer) energy transfer mechanism, combined with Heptamethine cyanine as the radiation near infrared light sensor, developed to the ratiometric method to quantify pH values and quantification of glutathione. (i) Heptamethine cyanine Based Semiconducting Polymer Dots for Detection of Cellular pH. The intracellular pH is very important for cell metabolism, and almost all proteins are based on pH values to maintain their structure and functionality. In addition, pH also plays a very important role in cell proliferation, apoptosis, drug resistance, phagocytosis, endocytosis and signaling. In addition, it has recently been shown that the cytosolic acidity of cells creatine repair is also important. So the intracellular pH is very strict, as long as there is a little abnormal on behalf of some physical problems, such as cancer and Alzheimer's disease, the greater the deviation may even cause death. Therefore, if we can accurately understand and monitoring the entire cell pH in vivo is a very important breakthrough. However, the traditional pH quantitative methods such as electrochemical method because of poor spatial resolution, NMR is limited because of operational difficulties, So we have to develop a faster and more convenient way to detect. Which by fluorescence detection technology, with high resolution, non-destructive, able to design to achieve high selectivity, is currently used in cell research. The ultimate goal of this fluorescent probe is to be able to achieve biological applications, so we use Heptamethine cyanine as a choice of dye, the use of its own structure with a long conjugate chain, modified with a electron-donating functional group based piperazine . The use of it in the acidic environment will be protonated, reducing the ability to donate electrons and extend the conjugate chain to absorb redshift phenomenon. Therefore, in this study, we use the advantage of Pdots easy - to - make the surface modification characteristics, in the polymer side we design of COOH functional groups, and in the near-infrared dye part is modified with NH2 functional groups. The FRET energy transfer mechanism was used to calibrate the pH value of the detective substance. Keywords : Semiconducting polymer dotsãNear-infrared dyesãpH sensingãFRETãFluorescence ratiometric analysis ( ii )Heptamethine cyanine Based Semiconducting Polymer Dots for cellular Quantification of Glutathione GSH is the most abundant bio-mercaptan in cells, the concentration range of 1~15 mmol / L. GSH is involved in many pathology and biology processes and plays an important role in maintaining the balance of intracellular redox states. ROS(reactive oxygen species), and different concentrations of GSH are also associated with a variety of cellular functions, including… Advisors/Committee Members: Ho-Hsiu Chou (chair), Yang-Hsiang Chan (committee member), Ming-Li Tsai (chair).

Subjects/Keywords: Azobenzene; Semiconducting polymer dots; Near-infrared dyes; pH sensing; FRET; Fluorescence ratiometric analysis; GSH sensing

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

LIN, D. (2017). Heptamethine Cyanine Based Semiconducting Polymer Dots for : ( i ) Detection of Cellular pH ; and ( ii ) Quantification of Glutathione. (Thesis). NSYSU. Retrieved from http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0806117-224007

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

LIN, DING-JYUN. “Heptamethine Cyanine Based Semiconducting Polymer Dots for : ( i ) Detection of Cellular pH ; and ( ii ) Quantification of Glutathione.” 2017. Thesis, NSYSU. Accessed September 22, 2017. http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0806117-224007.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

LIN, DING-JYUN. “Heptamethine Cyanine Based Semiconducting Polymer Dots for : ( i ) Detection of Cellular pH ; and ( ii ) Quantification of Glutathione.” 2017. Web. 22 Sep 2017.

Vancouver:

LIN D. Heptamethine Cyanine Based Semiconducting Polymer Dots for : ( i ) Detection of Cellular pH ; and ( ii ) Quantification of Glutathione. [Internet] [Thesis]. NSYSU; 2017. [cited 2017 Sep 22]. Available from: http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0806117-224007.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

LIN D. Heptamethine Cyanine Based Semiconducting Polymer Dots for : ( i ) Detection of Cellular pH ; and ( ii ) Quantification of Glutathione. [Thesis]. NSYSU; 2017. Available from: http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0806117-224007

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation


University of Illinois – Chicago

3. Kurilova, Svetlana P. Regulatory Roles of Cellular Diacylglycerol.

Degree: 2015, University of Illinois – Chicago

Kinetics of diacylglycerol and C1 domain interaction Lipids in biological membranes play significant roles in cellular signaling. Lipids vary not only in head groups but also in acyl chains. It has been reported that the acyl chain of lipids plays an important role in membrane curvature and signal transduction. In this work we investigated the importance of the degree of unsaturation in the acyl chain of diacylglycerol (DAG) in its interaction with the C1 domain. Fluorescence quenching assay showed that DAG with a highly unsaturated acyl chain binds more tightly to the C1 domain than DAG with saturated acyl chains. Moreover, our single molecule tracking studies using supported lipid bilayers as a model system and NIH 3T3 cells explain the kinetic aspect of that binding. Although more unsaturated DAG binds C1 domain tighter it dissociates from the membrane faster than in case of saturated DAG. This finding can explain why unsaturated lipids participate in cell signaling when saturated lipids mainly play a role of bulk lipids. This work provides the new inside about the dynamics of protein-lipid interaction. Extracellular function of diacylglycerol The heterogeneous distribution of lipid molecules in biological membranes depends on a specific region and situation. Selective distribution between bilayer leaflets can be achieved through diffusion (flip-flop) across membranes. DAG is one these lipids which is able to flip-flop rapidly because of a small, uncharged head group. Since the precise cellular localization of DAG determines its function and activity, it is important to understand its distribution and dynamics within a membrane leaflet and between inner and outer parts of cellular bilayers. We developed DAG sensor using mutagenesis and chemical labeling, which allowed accurate quantification of the lipid. Using this sensor we were able to detect very low concentration of DAG at the plasma membrane. It was found that ATP activation in NIH 3T3 cells elevates DAG in plasma membrane 5-7 times. Preliminary data suggest a significant basal level of DAG. It is known that there are two major sources of DAG: from phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and phosphatidylcholine (PC) and both can be initiated by ATP. Physiological roles of exofacial DAG remain unknown. It requires further investigation to understand the flip-flop process and its initiation. We discuss the novel four-color imaging with application of ratiometric analysis that can resolve these questions. Advisors/Committee Members: Cho, Wonhwa (advisor).

Subjects/Keywords: ratiometric analysis; diacylglycerol; DAG; lipid sensor; FCS; FRET; DAG sensor; lipid quantification; four color fluorescence imaging; in situ imaging; DAG kinetics; lipid-protein binding

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Kurilova, S. P. (2015). Regulatory Roles of Cellular Diacylglycerol. (Thesis). University of Illinois – Chicago. Retrieved from http://hdl.handle.net/10027/19806

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Kurilova, Svetlana P. “Regulatory Roles of Cellular Diacylglycerol.” 2015. Thesis, University of Illinois – Chicago. Accessed September 22, 2017. http://hdl.handle.net/10027/19806.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Kurilova, Svetlana P. “Regulatory Roles of Cellular Diacylglycerol.” 2015. Web. 22 Sep 2017.

Vancouver:

Kurilova SP. Regulatory Roles of Cellular Diacylglycerol. [Internet] [Thesis]. University of Illinois – Chicago; 2015. [cited 2017 Sep 22]. Available from: http://hdl.handle.net/10027/19806.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Kurilova SP. Regulatory Roles of Cellular Diacylglycerol. [Thesis]. University of Illinois – Chicago; 2015. Available from: http://hdl.handle.net/10027/19806

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

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