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HTPQA熒光探針 優(yōu)點(diǎn)：
HTPQA熒光探針 是一種酶熒光成像探針，實(shí)現(xiàn)了活細(xì)胞內(nèi)堿性磷酸酶活性的原位成像檢測(cè)。該合成的堿性磷酸酶探針具有靈敏度高、選擇性好等優(yōu)點(diǎn)。該探針能夠解決傳統(tǒng)水溶性探針信號(hào)易擴(kuò)散、衰減的問題，從而實(shí)現(xiàn)堿性磷酸酶的原位成像檢測(cè)，除此之外，還成功應(yīng)用于成像檢測(cè)骨肉瘤細(xì)胞及組織中堿性磷酸酶的活性。這一新型固態(tài)發(fā)光成像探針的開發(fā)呈現(xiàn)出固態(tài)發(fā)光熒光染料在蛋白酶原位成像檢測(cè)中的巨大優(yōu)勢(shì)。

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Enzyme-responsive, fluorogenic probes fail to provide in situ information because the released fluorophores tend to diffuse away from the reaction sites. The problem of diffusive signal dilution can be addressed by designing a probe that upon enzyme conversion releases a fluorophore that precipitates. An excited-state intramolecular proton transfer (ESIPT)-based solid-state fluorophore HTPQ was developed that is strictly insoluble in water and emits intense fluorescence in the solid state, with λex/em=410/550 nm, thus making it far better suited to use with a commercial confocal microscope. HTPQ was further utilized in the design of an enzyme-responsive, fluorogenic probe (HTPQA), targeting alkaline phosphatase (ALP) as a model enzyme. HTPQA makes possible diffusion-resistant in situ detection of endogenous ALP in live cells. It was also employed in the visualizing of different levels of ALP in osteosarcoma cells and tissue, thus demonstrating its interest for the diagnosis of this type of cancer.

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       目標(biāo)探針HTPQA（紅色信號(hào)通道）與商品化ALP探針4-MUP在HeLa細(xì)胞中共同孵育的成像圖。從成像結(jié)果可以看出：（A, B）HTPQA能夠?qū)崿F(xiàn)ALP的原位檢測(cè)，而4-MUP則擴(kuò)散至整個(gè)細(xì)胞質(zhì)；（C, D）HTPQA信號(hào)穩(wěn)定，長(zhǎng)時(shí)間監(jiān)測(cè)信號(hào)無明顯擴(kuò)散，而4-MUP的信號(hào)逐漸消失。

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