| 摘要: | 本論文以實驗室先前研究多光子染料的經驗，將研究方向延伸應用在開發雙光子螢光探針。從分子設計開始，再透過有機合成，經光譜鑑定結構過後，確認目標的螢光探針是否成功被合成出。接續量測線性光學，得到吸收光譜、螢光光譜和螢光量子產率等基本資訊。再測量非線性光學的Photo excitation和Power dependence實驗，以取得雙光子吸收的數據。最後進行細胞實驗，來觀察所開發出的螢光探針實際在細胞內的表現為何。
接下來以結構構成來將螢光探針分成兩個系列。分別是第一系列：「以Fluorene作為π-bridge」之兩個螢光探針，同時會對四種結構相仿的化合物進行光學探討；以及第二系列：「將Benzothiazole官能基化」之三個螢光探針，也會對結構相仿的四種化合物進行光學探討。共計五個螢光探針，包含了四個標定溶酶體之螢光探針，和一個標定細胞膜之螢光探針。
第一系列的兩個螢光探針有成功合成並經光譜鑑定確認，且已經做了初步的細胞實驗，確定可以染上細胞且被影像紀錄，後續的實驗結果將會逐一補齊。在經過光學實驗的量測後，於Donor部分將一級胺的NH2修飾成三級胺的Morpholine之後，亮度明顯下降。而Acceptor部分，Benzothiazole 的亮度表現比起[1,2,4]Triazolo[1,5-a]pyridine來的更為出色。
第二系列的三個螢光探針一樣有成功合成並經光譜鑑定確認，並且做了初步的細胞實驗，確定可以染上細胞且被影像紀錄，後續的實驗結果也將一併補上。在經過光學實驗的量測後，相比未官能基化的模型分子，官能基化後的分子整體亮度有所下降。其中下降的幅度又以連接Benzothiazole的原子為「氮」的探針比連接的原子為「氧」的探針更多，但亮度還是足以使用的。
;This thesis extends the research direction to the development of two photon fluorescent probes based on the laboratory′s previous experience in researching multi-photon dyes. First, we started with molecular design, and synthesized from organic synthesis. It was confirmed whether the fluorescent probe had been correctly synthesized by spectrum. Next step was measured the linear optics to obtain optical properties such as absorption spectrum, emission spectrum and quantum yield. After that, we measured the Photo excitation and Power dependence experiments of non-linear optics from two-photon absorption. Finally, cell experiments were performed to observe how would the fluorescent probes worked in live cells.
The fluorescent probes are divided into two series by structural composition. The first series is two of the fluorescent probes composed of the "Fluorene as π-bridge" unit. Furthermore, we would do the optical experiments of four similar compounds. And the second series is three of
fluorescent probes, which are "functionalization of Benzothiazole." Then, we would also do the optical experiments of four similar compounds. There are five fluorescent probes in total, including four lysosome targeted fluorescent probes and one for cell membrane-targeted.
The first series of two fluorescent probes have been successfully synthesized and confirmed by spectral identification. Cell experiment indicate that the cells can be stained with these probes, it could be recorded by photoing at the same time. The other unfinished cell experimental will be completed in the future. In addition, we can check the fact from the optical experiment. When the donor part converted from the NH2 of primary amine to morpholine of the tertiary amine, the brightness decreases obviously. On the other side of the acceptor part, the brightness performance of Benzothiazole is much better than
[1,2,4]Triazolo[1,5-a]pyridine.
The second series of three fluorescent probes have been also successfully synthesized and confirmed by spectral identification. Cell experiment indicate that the cells can be stained with these probes, it could be recorded by photoing as well. The other unfinished cell experimental will be completed in the future. Compared with the non functionalized model molecule, the brightness of the functionalized molecule decreased through measurement by optical experiment. The probe with the atom attaches to the benzothiazole as "nitrogen" is darker than the probe with the atom as the "oxygen" attached. However, the brightness is still enough to be used. |
