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熒光淬滅又稱熒光熄滅或萃滅,是指導(dǎo)致特定物質(zhì)的熒光強(qiáng)度和壽命減少的所有現(xiàn)象。引起熒光淬滅的物質(zhì)稱為熒光淬滅劑。SouthernBiotech專門開發(fā)的Fluoromount-G系列熒光封片劑是以甘油為基礎(chǔ),加入抗熒光淬滅劑,可明顯降低熒光淬滅現(xiàn)象,提高樣本中熒光信號(hào)的穩(wěn)定性和持久性。
Fluoromount-G®(貨號(hào):0100-01)
Fluoromount-G®是一種水溶性封片劑,用來對(duì)最后一步染色為水溶液體系的玻片樣本進(jìn)行封片,可以明顯降低熒光顯微鏡下玻片樣本分析中的熒光淬滅現(xiàn)象。據(jù)報(bào)道,F(xiàn)luoromount-G®的折射率約為1.4。Fluoromount-G®已有超過20,000多篇文獻(xiàn)引用,與100多種熒光染料和試劑兼容。具體詳情,請(qǐng)聯(lián)系SouthernBiotech代理商欣博盛生物!
圖1.用抗BCAS2抗體和抗PLZF抗體對(duì)小鼠P8睪丸的石蠟切片進(jìn)行染色。用Hoechst 33342對(duì)DNA進(jìn)行染色,并用Fluoromount-G®(貨號(hào)0100-01)封片。圖片來自Liu W , Wang F , Xu Q, Shi J , Zhang X , Liu X , et al . BCAS2 is involved in alternative mRNA splicing in spermatogonia and the transition to meiosis. Nat Commun. 2017; 8: 14182.
圖2. 用AF488標(biāo)記的鬼筆環(huán)肽對(duì)卷身羅斯考夫蠕蟲胚胎全標(biāo)本進(jìn)行染色,并用Fluoromount-G®(貨號(hào)0100-01)封片。圖片來自Semmler H, Bailly X, Wanninger A. Myogenesis in the basal bilaterian Symsagittifera roscoffensis (Acoela). Front Zool.2008; 5:14.
圖3. 用anti-Zpr3抗體對(duì)冰凍斑馬魚視網(wǎng)膜切片進(jìn)行染色,然后用AF543二抗和MBODEC染色并用Fluoromount-G®(貨號(hào)0100-01)封片。圖片來自Watanabe K ,Nishimura Y, Oka T, Nomoto T, Kon T, Shitou T,et al. In vivo imaging of zebrafish retinal cells using fluorescent coumarin derivatives. BMC Neurosci.2010; 11:116.
DAPI Fluoromount-G®(貨號(hào):0100-20)
DAPI Fluoromount-G®是一種水溶性的、含DAPI的化合物,用來對(duì)最后一步染色是在水溶液體系中完成的玻片樣本進(jìn)行封片。當(dāng)與DNA結(jié)合時(shí),DAPI會(huì)發(fā)出熒光,通常用作細(xì)胞核/染色體復(fù)染物。由于DAPI Fluoromount-G®是水溶性的,可以通過將玻片浸入PBS溶液中直到蓋玻片松開來移除蓋玻片。
圖4.HCC827細(xì)胞先用抗E-鈣粘蛋白抗體染色,隨后用二抗染色,最后用DAPI Fluoromount-G®(貨號(hào)0100-20)封片。圖片來自Narita M, Shimura E, Nagasawa A , Aiuchi T , Suda Y ,Hamada Y et al . Chronic treatment of non-small-cell lung cancer cells with gefitinib leads to an epigenetic loss of epithelial properties associated with reductions in microRNA-155 and -200c. PLOS One.2017; 12 (2): e0172115.
圖5.在無血清培養(yǎng)基中生長(zhǎng)的人中性粒細(xì)胞用抗中性粒細(xì)胞彈性蛋白酶抗體染色,然后用AF555偶聯(lián)的二抗染色,并用DAPI Fluoromount-G®(貨號(hào)0100-20)封片。圖片來自Kamoshida G, Kikuchi-Ueda T, Nishida S, Tansho-Nagakawa S , Kikuchi H, Ubagai T , et al . Spontaneous formation of neutrophil extracellular traps in serum-free culture conditions. FEBS Open Bio. 2017; 7: 877-86.
Fluoromount-G® Anti-Fade(貨號(hào):0100-35)
Fluoromount-G®Anti-Fade是一種非硬化的水基化合物,用于顯微鏡應(yīng)用中對(duì)熒光強(qiáng)度保持至關(guān)重要的載玻片的封片,適用于石蠟切片、冰凍切片及細(xì)胞樣本。Fluoromount-G® Anti-Fade含有專有的防淬滅成分,可在分析過程中抑制光漂白,非常適合熒光強(qiáng)度至關(guān)重要的顯微鏡應(yīng)用。
圖6.冷凍新生小鼠軟骨切片用正常兔血清封閉,用小鼠抗II型膠原蛋白抗體染色,隨后用BIOT標(biāo)記的兔抗小鼠IgG(H+L)抗體(貨號(hào):6125-08)、FITC標(biāo)記的鏈霉親和素(貨號(hào):7100-02)、DAPI染色,最后用Fluoromount-G® Anti-Fade(貨號(hào):0100-35)封片。
圖7. 用小鼠抗人CD44-UNLB(貨號(hào)9400-01)對(duì)人胰腺癌細(xì)胞系MIA-Paca-2染色,然后用山羊抗小鼠IgG,Human ads-BIOT(貨號(hào)1030-08)和鏈霉親和素-CY3(貨號(hào)7100-12)染色。
SouthernBiotech的Fluoromount-G系列熒光封片劑具有半永-久封固性,可用作長(zhǎng)期保存玻片樣本的制作。
產(chǎn)品信息
貨號(hào) | 產(chǎn)品名稱 | 規(guī)格 |
0100-01 | Fluoromount-G® | 25 mL |
0100-20 | DAPI Fluoromount-G® | 20 mL |
0100-35 | Fluoromount-G® Anti-Fade | 20 mL |
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