細胞內比例型pH感應的新奈米設計材料

New Design Nanomaterials for Ratiometric Intracellular pH Sensing

菲比(Feby Wijaya Pratiwi)

指導教授 ： 陳培菱

共同指導教授 ： 牟中原(CHUNG-YUAN MOU)

國立臺灣大學/理學院/化學研究所/博士(2018年)

https://doi.org/10.6342/NTU201801602

摘要

我們發展兩個全新的細胞內pH感應探針設計，利用可行的奈米材料，量子點以及中孔洞奈米矽球。我們設計兩個奈米點在pHi感應探針上. 首先，我們在二個射極端合成參雜錳二價離子的單一量子點，有顯示二個螢光團，表示常見的比例型pH感應器是可靠的. 將二個簡化至一個的比例型感應器設計可促使影像等的相關應用. 當HeLa細胞再進行細胞吞噬作用時，用具有生物相容性的DPA-MnQdots來長時間監測微區域的環境pH值. 接著，我們合成中孔洞奈米矽球，並將三個不同染劑的含肽溶酶體載入於中孔洞奈米矽球. 此方法可以使比例型pH達到長時間可靠，其精準度至0.3pH單位，且擴大pH檢測的範圍. 由於這有良好的亮度及穩定度，該材料符合單分子追蹤探針的標準. 最後，我們發展三維單分子追蹤技術，監控靶向溶酶體在FORMSN周圍區域即時的pH變化.

關鍵字

細胞內pH感應 ； 量子點 ； 中孔洞奈米矽球 ； 單分子追蹤

並列摘要

We developed new design intracellular pH (pHi) sensing probes by using available nanomaterials, quantum dots and Mesoporous Silica Nanoparticles. First, we synthesized single Mn2+doped quantum dots (DPA-MnQdots) which exhibit 2 emissions and show reliable and general features of common ratiometric sensing based on 2 fluorophores. The changing from “two” to “one” will simplify ratiometric sensing design and promote the corresponding applications like imaging. The biocompatible DPA-MnQdots were applied for long-term monitoring of local microenvironments pH values during endocytosis in HeLa cells. Second, we synthesis dye-loaded fluorescent mesoporous silica nanoparticles decorated lysosomal sorting peptide (YXXɸ), called peptide(YXXɸ)-FORMSN, potentially act as drug carrier to treat Lysosomal Storage Diseases as well as pHi sensing probe. This approach allowed long-term reliable ratiometric pH measurements with precision better than 0.3 pH units and broaden the window of pH value detection. Owing to its excellent brightness and stability, the FORMSN could meet the criteria to serve as a single particle tracking (SPT) probe. In addition, super-resolution 3D SPT with a simple marker-assisted drift correction, was also developed to visualize the motion and record the local pH value of individual peptide-FORMSN during its journey with high-precision. The overall data from fluctuation of local pH value surrounding of peptide-FORMSNs, real-time trajectory, and real-time speed variation represent the dynamic interactions of peptide-FORMSNs with their cellular hosts during their entire journey. As result, we were able to construct the whole pathway, traditional and alternative route, of peptide -FORMSNs for targeting lysosome.

並列關鍵字

Intracellular pH sensing ； quantum dots ； Mesoporous Silica Nanoparticles ； Single Particle Tracking

參考文獻

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