Fluoro-Gold can be injected using several different methods, including pressure, iontophoretic and otherapplications developed by a variety of researchers. See Schmued and Fallon, Fluoro-Gold: "A

fluorescent retrograde axonal tracer with numerous unique properties", Brain Research, 377 (1986) 147- 154 as well as Pieribone and Aston-Jones, "The Iontophoretic Application of Fluoro-Gold for the study ofaf erents to deep brain nuclei", Brain Research, 475 (1988) 259-271. Many researchers have developedtheir own modified procedures. Use of the antibody should not be dependent upon the methodology usedto employ Fluoro-Gold.

After the Fluoro-Gold has been injected, floating sections (we used thirty um sections from a rat perfusedwith 4% formaldehyde) are incubated with the Fluoro-Gold Antibody solution overnight at 4o C. Sectionsare washed, then incubated in Biotinylated GAR (Vector Labs) at 1/1000 for 1 hour at room temperatureand washed again. Sections are then incubated in Avidin/Biotin (Vector Labs) at 1/1000 for 1 hour, washed and transferred to Diaminobenzidine (.04%) and Nickel Chloride (2.5%) in 0.1 M NaAcetate with0.06% H2O2 for six minutes. Sections are then washed, mounted, dried, dehydrated and cover slipped.

It has been our experience that if stored and prepared in the manner set out above, each vial of the antibody should treat 300 to 1000 thirty um sections of the albino rat brain (or similar sized animals) ifyou use the Vector elite kit. However, you should experiment with the concentration and procedures todetermine which best fit your circumstance.

艾美捷Fluorochorome熒光金(Fluoro-Gold)抗體相關(guān)介紹：

氟金可以用幾種不同的方法注射，包括壓力、離子電滲和各種研究人員開發(fā)的治療方法。參見Schmoud和Fallon，F(xiàn)luoro Gold：“A熒光逆行軸突示蹤劑具有許多獨(dú)-特的財(cái)產(chǎn)"，《大腦研究》，377（1986）147-154，以及Pieribone和Aston-Jones，“熒光金在研究腦深部細(xì)胞核的遺傳物質(zhì)中的離子導(dǎo)入應(yīng)用"，《腦研究》，475（1988）259-271。許多研究人員開發(fā)了他們自己的改進(jìn)程序?？贵w的使用不應(yīng)依賴于使用熒光金的方法。

注射Fluoro Gold后，將漂浮切片（我們使用了灌注了4%甲醛的大鼠的30μm切片）與Fluoro金抗體溶液在4℃下孵育過夜。切片被洗滌，然后在室溫下在生-物-素化GAR（Vector Labs）中以1/1000孵育1小時(shí)，然后再次洗滌。然后將切片在Avidin/Biotin（Vector Labs）中以1/1000孵育1小時(shí)，洗滌并轉(zhuǎn)移至二氨基聯(lián)苯胺（.04%）和氯化鎳（2.5%）中的0.1M乙酸鈉和0.06%H2O2中6分鐘。然后清洗、安裝、干燥、脫水并蓋上蓋子。

根據(jù)我們的經(jīng)驗(yàn)，如果按照上述方式儲存和制備，如果您使用Vector精英試劑盒，每小瓶抗體應(yīng)處理300至1000個(gè)30μm的白化大鼠腦（或類似大小的動物）切片。然而，你應(yīng)該嘗試集中注意力和程序，以確定哪一個(gè)最-適合你的環(huán)境。

Fluorochorome熒光金(Fluoro-Gold)抗體的檢查和攝影：

使用寬帶紫外（UV）激發(fā)濾光器，熒光顯微鏡觀察熒光金。使用與在寬帶紫外（如真藍(lán)、快藍(lán)、核黃）下激發(fā)的其他熒光逆行示蹤劑相同的濾鏡組，并應(yīng)特別為熒光顯微鏡甘油或水制作物鏡。由于塑料確實(shí)吸收紫外線，因此不建議通過塑料培養(yǎng)皿等進(jìn)行觀察。推薦使用T-Max（柯達(dá)，黑白）和Ektachrome 200（柯達(dá)，彩色幻燈片）。曝光時(shí)間通常從20秒到1.5分鐘不等。

熒光金(Fluoro-Gold)抗體根據(jù)不同規(guī)格分為單管和二管：

熒光金抗體-單管/100ul（FC20001）

熒光金抗體-2管/2x100ul（FC20002）