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Detection and Interpretation of Fluorescence Signals Generated by Excitable Cells and Tissues

機(jī)譯：興奮細(xì)胞和組織產(chǎn)生的熒光信號(hào)的檢測(cè)和解釋

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Part 1: High-Sensitivity Amplifiers for Detecting Fluorescence .;Monitoring electrical activity and Cai 2+ transients in biological tissues and individual cells increasingly utilizes optical sensors based on voltage-dependent and Cai 2+-dependent fluorescent dyes. However, achieving satisfactory signal-to-noise ratios (SNR) often requires increased illumination intensities and/or dye concentrations, which results in photo-toxicity, photo-bleaching and other adverse effects limiting the utility of optical recordings. The most challenging are the recordings from individual cardiac myocytes and neurons. Here we demonstrate that by optimizing a conventional transimpedance topology one can achieve a 10-20 fold increase of sensitivity with photodiode-based recording systems (dependent on application). We provide a detailed comparative analysis of the dynamic and noise characteristics of different transimpedance amplifier topologies as well as the example(s) of their practical implementation.;Part 2: Light-Scattering Models for Interpretation of Fluorescence Data.;Current interest in understanding light transport in cardiac tissue has been motivated in part by increased use of voltage-sensitive and Ca i2+-sensitive fluorescent probes to map electrical impulse propagation and Cai2+-transients in the heart. The fluorescent signals are recorded using such probes represent contributions from different layers of myocardial tissue and are greatly affected by light scattering. The interpretation of these signals thus requires deconvolution which would not be possible without detailed models of light transport in the respective tissue. Which involves the experimental measurements of the absorption, scattering, and anisotropy coefficients, mua, mu s, and g respectively.;The aim of the second part of our thesis was to derive a new method for deriving these parameters from high spatial resolution measurements of forward-directed flux (FDF). To this end, we carried out high spatial resolution measurements of forward-directed flux (FDF) in intact and homogenized cardiac tissue, as well as in intralipid-based tissue phantoms. We demonstrated that in the vicinity of the illuminated surface, the FDF consistently manifested a fast decaying exponent with a space constant comparable to the decay rate of ballistic photons. Using a Monte Carlo model we obtained a simple empirical formula linking the rate of the fast exponent to the scattering coefficient, the anisotropy parameter g, and the numerical aperture of the probe. The estimates of scattering coefficient based on this formula were validated in tissue phantoms. The advantages of the new method are its simplicity and low-cost.
機(jī)譯：第1部分：用于檢測(cè)熒光的高靈敏度放大器。監(jiān)測(cè)生物組織和單個(gè)細(xì)胞中的電活動(dòng)和Cai 2+瞬態(tài)的方法越來越多地使用基于電壓依賴性和Cai 2+依賴性熒光染料的光學(xué)傳感器。但是，要獲得令人滿意的信噪比（SNR），通常需要增加照明強(qiáng)度和/或染料濃度，這會(huì)導(dǎo)致光毒性，光漂白和其他不利影響，限制了光學(xué)記錄的實(shí)用性。最具挑戰(zhàn)性的是單個(gè)心臟心肌細(xì)胞和神經(jīng)元的錄音。在這里，我們證明了通過優(yōu)化傳統(tǒng)的跨阻拓?fù)?，可以使基于光電二極管的記錄系統(tǒng)的靈敏度提高10到20倍（取決于應(yīng)用）。我們提供了不同互阻放大器拓?fù)涞膭?dòng)態(tài)和噪聲特性的詳細(xì)比較分析，以及它們的實(shí)際實(shí)現(xiàn)示例。；第2部分：用于解釋熒光數(shù)據(jù)的光散射模型。當(dāng)前對(duì)理解光的興趣心臟組織中的轉(zhuǎn)運(yùn)在一定程度上是由于越來越多地使用電壓敏感和Ca i2 +敏感的熒光探針來繪制心臟中的電脈沖傳播和Cai2 +瞬變的圖。使用這種探針記錄的熒光信號(hào)代表了心肌組織不同層的貢獻(xiàn)，并且受光散射的影響很大。因此，這些信號(hào)的解釋需要去卷積，如果沒有相應(yīng)組織中光傳輸?shù)脑敿?xì)模型，這將是不可能的。其中涉及吸收，散射和各向異性系數(shù)分別為mua，mu s和g的實(shí)驗(yàn)測(cè)量。本論文第二部分的目的是推導(dǎo)一種從高空間分辨率測(cè)量推導(dǎo)這些參數(shù)的新方法。正向通量（FDF）。為此，我們?cè)谕暾途|(zhì)的心臟組織以及基于脂質(zhì)內(nèi)的組織模型中對(duì)前向通量（FDF）進(jìn)行了高空間分辨率的測(cè)量。我們證明，在受照表面附近，F(xiàn)DF始終表現(xiàn)出快速衰減指數(shù)，其空間常數(shù)可與彈道光子的衰減率相媲美。使用蒙特卡洛模型，我們獲得了一個(gè)簡(jiǎn)單的經(jīng)驗(yàn)公式，該公式將快速指數(shù)的速率與散射系數(shù)，各向異性參數(shù)g和探針的數(shù)值孔徑聯(lián)系起來。基于此公式的散射系數(shù)估計(jì)值已在組織體模中得到驗(yàn)證。新方法的優(yōu)點(diǎn)是它的簡(jiǎn)單性和低成本。

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  • 作者

    Costantino, Anthony J.;

  • 作者單位

    State University of New York at Binghamton.;

  • 授予單位 State University of New York at Binghamton.;
  • 學(xué)科 Electrical engineering.;Optics.;Biomedical engineering.
  • 學(xué)位 Ph.D.
  • 年度 2017
  • 頁碼 269 p.
  • 總頁數(shù) 269
  • 原文格式 PDF
  • 正文語種 eng
  • 中圖分類 水產(chǎn)、漁業(yè);
  • 關(guān)鍵詞

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