發(fā)布時(shí)間：2018-03-23 23:39

  本文選題：熒光探針　切入點(diǎn)：雙光子熒光　出處：《蘭州大學(xué)》2017年碩士論文

【摘要】：雙光子吸收技術(shù)自問(wèn)世以來(lái)一直受到了廣泛的關(guān)注。與單光子吸收材料相比,雙光子吸收材料在分辨率、穿透深度具有顯著的優(yōu)勢(shì),可以用于顯微成像、微紡織技術(shù)、三維數(shù)據(jù)存儲(chǔ)、光限幅、上轉(zhuǎn)換發(fā)光、光動(dòng)力學(xué)治療以及藥物靶向釋放等諸多領(lǐng)域。特別是雙光子顯微技術(shù),以近紅外的激光為光源對(duì)生物樣品進(jìn)行成像,具有穿透性強(qiáng),空間分辨率高,背景熒光干擾小,以及對(duì)生物樣品的光損傷較小等優(yōu)點(diǎn),在生物醫(yī)學(xué)領(lǐng)域具有廣闊的應(yīng)用前景。然而,傳統(tǒng)的雙光子材料常常具有大共軛結(jié)構(gòu),水溶性差、細(xì)胞穿透能力差、生物毒性也較大,并不適用于生物成像。因此,設(shè)計(jì)合成具有較高雙光子吸收截面的有機(jī)小分子用于生物體內(nèi)細(xì)胞、血管、組織成像,具有重要的研究?jī)r(jià)值。本文設(shè)計(jì)合成了兩種具有雙光子吸收特性的熒光小分子,對(duì)其發(fā)光性能進(jìn)行了系統(tǒng)的研究,探索它們?cè)谏锍上裰械膽?yīng)用。具體的研究?jī)?nèi)容包括:1、設(shè)計(jì)合成了一類以寡聚苯乙烯為骨架的雙光子次氯酸熒光探針OPV-HOCl,并將其應(yīng)用于活細(xì)胞及組織內(nèi)的雙光子成像。在寡聚苯乙烯骨架上引入次氯酸識(shí)別基團(tuán)——氧硫雜環(huán)戊烷,通過(guò)1H-NMR、13C-NMR、HRMS對(duì)其結(jié)構(gòu)進(jìn)行了表征,并通過(guò)紫外光譜、熒光光譜等進(jìn)一步研究了該探針對(duì)次氯酸的響應(yīng)性能,測(cè)定了其雙光子吸收截面。加入次氯酸以后,探針?lè)肿幽┒说难趿螂s環(huán)戊烷基團(tuán)被氧化,并生成醛基。由于分子內(nèi)強(qiáng)烈的電荷轉(zhuǎn)移導(dǎo)致產(chǎn)物的雙光子吸收截面提高了近15倍(從78.9 GM提高到1131.5 GM),因此OPV-HOCl可以作為一個(gè)雙光子“turn-on”型次氯酸熒光探針。此外,該探針還具有反應(yīng)速度快、選擇性好、pH適用范圍寬等優(yōu)點(diǎn)。MTT實(shí)驗(yàn)表明該探針具有較小的細(xì)胞毒性。由于該探針優(yōu)異的次氯酸響應(yīng)性能和較小的生物毒性,我們成功地將其用于小鼠膠質(zhì)瘤細(xì)胞BV-2中次氯酸的檢測(cè),研究表明該探針可以透過(guò)細(xì)胞膜,并對(duì)細(xì)胞中外源性次氯酸和脂多糖誘導(dǎo)產(chǎn)生的內(nèi)源性次氯酸具有高選擇性的快速響應(yīng)。2、合成了具有雙光子吸收性質(zhì)的吲哚四方酸染料SQA-Br,并將其負(fù)載于微米級(jí)的聚苯乙烯微球中,用于腦中風(fēng)區(qū)域的檢測(cè)研究。研究表明,聚苯乙烯微球可以為染料提供相對(duì)惰性的環(huán)境,避免了聚集誘導(dǎo)熒光猝滅,使得染料在水溶液中仍具有良好的發(fā)光性能,并且染料在微球中的熒光壽命和量子產(chǎn)率與溶液中相比均有一定程度的提高,因此在生物成像中具有潛在的應(yīng)用價(jià)值。我們將負(fù)載四方酸的聚苯乙烯微球注射進(jìn)入中風(fēng)小鼠體內(nèi),發(fā)現(xiàn)微球從中風(fēng)損傷區(qū)域擴(kuò)散出去并在周?chē)鷧^(qū)域富集,可以用于中風(fēng)損傷區(qū)域的初步識(shí)別。
[Abstract]:Two-photon absorption technology has been paid more and more attention since it came into being. Compared with single-photon absorption material, two-photon absorption material has obvious advantages in resolution and penetration depth, so it can be used in micro-imaging and micro-textile technology. Three dimensional data storage, optical limiting, up-conversion luminescence, photodynamic therapy and drug targeted release, etc., especially two-photon microscopy, imaging biological samples with near-infrared laser as light source, has strong penetrability. Because of its advantages of high spatial resolution, low background fluorescence interference and low light damage to biological samples, it has broad application prospects in biomedical fields. However, traditional two-photon materials often have large conjugate structures and poor water solubility. Therefore, organic small molecules with high two-photon absorption cross sections are designed and synthesized for biological imaging of cells, blood vessels and tissues. In this paper, two kinds of fluorescent small molecules with two-photon absorption characteristics are designed and synthesized, and their luminescence properties are systematically studied. The specific research contents include: 1, a two-photon hypochlorite fluorescence probe OPV-HOCl with oligopolystyrene as skeleton was designed and synthesized, and applied to two-photon formation in living cells and tissues. Like. Introduction of hypochlorite recognition group-oxo-thiocyclopentane on the framework of oligopolystyrene, The structure of the probe was characterized by 1H-NMR-13C-NMR-HRMS. The response of the probe to hypochloric acid was further studied by UV and fluorescence spectra. The two-photon absorption cross section of the probe was determined after the addition of hypochloric acid. The oxygen-thiocyclopentane group at the end of the probe was oxidized. Due to the strong intramolecular charge transfer, the two-photon absorption cross section of the product was increased by nearly 15 times (from 78.9 GM to 1131.5 GMN), so OPV-HOCl can be used as a two-photon "turn-on" type hypochloric acid fluorescence probe. MTT assay showed that the probe had less cytotoxicity because of its excellent hypochloric acid response and low biotoxicity. We have successfully applied it to the detection of hypochloric acid in mouse glioma cell line BV-2. The results show that the probe can penetrate the cell membrane. And the endogenous hypochloric acid induced by exogenous hypochloric acid and lipopolysaccharide was highly selective and rapid response. SQA-Br2, a two-photon absorption dye, was synthesized and loaded into micrometer polystyrene microspheres. The results show that polystyrene microspheres can provide a relatively inert environment for dyes, avoid agglomeration induced fluorescence quenching, and make the dyes still have good luminescence properties in aqueous solution. Moreover, the fluorescence lifetime and quantum yield of dyes in the microspheres were improved to a certain extent compared with those in the solution. We injected polystyrene microspheres loaded with tetragonal acid into apoplectic mice and found that the microspheres spread out from the wind-damaged area and enriched in the surrounding area. It can be used to identify the area of stroke injury.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O657.3

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