綠色螢光蛋白發光團，從早期的發現、放光機制並類似物合成的探討，時至今日有越來越多在生物標記、螢光化學感測器上的應用。因diol基團在文獻中有與金屬離子結合的記載，所以我們利用glycidol將其引入鄰位胺基的綠色螢光蛋白類似物 (o-ABDI)，得到化合物o-DHPBDI去探討其偵測金屬離子之性質。
從實驗結果我們發現o-DHPBDI能感測到特定金屬離子Cu2+、Cr3+、Fe3+，於吸收光譜中有isosbestic point出現；並且在不同溶劑中會因化合物的free-form與complex的比例不同而放出不同顏色的螢光。從modified Stern-Volmer equation中我們得到o-DHPBDI對Cr3+、Fe3+之結合常數約落在104；Job’s plot的實驗結果則是呈現o-DHPBDI與Cr3+、Fe3+皆為1:1的鍵結比例。對於o-DHPBDI與金屬離子可逆性之探討，我們發現錯合物在滴定EDTA後，EDTA能將Cr3+、Fe3+從錯合物中抓回，即o-DHPBDI為一可逆的化學感測器。此外，在錯合物養晶的過程中，我們也發現o-DHPBDI會與Cu2+進行離子催化反應形成合環的產物。
在Astrocyte的細胞實驗中，我們發現o-DHPBDI雖可以通過細胞進行染色，於共軛焦顯微鏡下呈現綠色顆粒螢光，但其放出的螢光強度不是太好，所以我們對其結構進行修飾得到化合物o-PDABDI希望藉此提高化合物進入細胞的數量，達到增強螢光的目的，從結果也可以看到綠色的螢光確實有增多的趨勢。對此初步的實驗結果，我們猜測這應該是化合物與離子或是離子-蛋白質巨分子之複合體結合所放出的螢光。

We synthesis an ortho analogue of GFP chromophore (o-DHPBDI) which contains diol moiety, this compound can detect particular cations, such as Cr3+ and Fe3+, resulting in fluorescent emission at approximately 550nm decrease, while 430nm increase, these two fluorescence intensity and wavelength will effected by different solvents. We indicate the dual emmsion is caused by the ration of free-form and complex. Furthermore, according to X-ray single crystal, we found that o-DHPBDI will catalyzed by copper(II) forming annulation product.
In this biological experiment, we use o-DHPBDI to dye the Astrocyte. although o-DHPBDI can go through the cell membrane and display the green fluorescence, its fluorescence intensity is relatively low. To improve this problem, we use acetic anhydride to get acetylated product (o-PDABDI) which is modifying by o-DHPBDI. To our delight, fluorescence intensity is relatively stronger. We postulate that fluorescence displayed inside the Astrocyte is casued by the binding of compound with ions or ion-proteins complex.

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