熒光適配器用于樣品的預(yù)篩選
美國(guó)NIGHTSEA SFA-RB體視顯微鏡熒光適配器可以將您的常規(guī)實(shí)驗(yàn)室體視顯微鏡變成一種有價(jià)值的工具,用于在進(jìn)入更高分辨率的熒光顯微鏡或共聚焦顯微鏡之前預(yù)先篩選樣品制備的熒光。上海峰志儀器有限公司代理銷售美國(guó)nightsea體視顯微鏡熒光適配器和熒光手電筒。有關(guān)體視顯微鏡熒光適配器產(chǎn)品介紹請(qǐng)瀏覽《美國(guó)nightsea熒光適配器 》,上海峰志儀器有限公司銷售的熒光觀察設(shè)備能夠觀察植物愈傷、葉片、種子、根系等上面的轉(zhuǎn)基因的表達(dá),還能觀察轉(zhuǎn)基因在動(dòng)物如老鼠、斑馬魚、果蠅等實(shí)驗(yàn)動(dòng)物上的表達(dá)。有便攜式單波長(zhǎng)熒光手電筒,也有手持式大面積照射高強(qiáng)度的雙波長(zhǎng)激發(fā)光源,上海峰志提供免費(fèi)試機(jī),有興趣的老師和同學(xué)可按照網(wǎng)頁底部聯(lián)系聯(lián)系。
挑戰(zhàn)
生物樣品的高分辨率成像主要基于熒光技術(shù)。共聚焦、雙光子和高分辨率復(fù)合熒光顯微鏡幾乎總是有限的資源。它們通常僅位于成像核心設(shè)施中,并且可按計(jì)劃按使用付費(fèi)。將熒光團(tuán)引入樣品的過程并不總是成功的。染色,將含GFP的質(zhì)粒引入細(xì)胞,免疫組織化學(xué) - 這些都是錯(cuò)誤的?;〞r(shí)間在高端系統(tǒng)上尋找熒光并不罕見,而那里甚至沒有發(fā)現(xiàn)任何熒光。
實(shí)用的解決方案
美國(guó)NIGHTSEA SFA-RB可以在標(biāo)準(zhǔn)體視顯微鏡上對(duì)標(biāo)本進(jìn)行熒光預(yù)篩選。您看到的細(xì)節(jié)并不重要 - 熒光的簡(jiǎn)單存在或不存在以及一般位置可以讓您知道是否值得將標(biāo)本帶到成像核心。在使用費(fèi)的直接費(fèi)用和浪費(fèi)在查看非熒光樣品的時(shí)間之間,NIGHTSEA系統(tǒng)不需要很多節(jié)省的旅行來支付自己。
一位研究人員的工作要求用Alexa Fluor 488 Phalloidin染色兔子腰大肌纖維。對(duì)于沒有沾染污漬的樣品,有些令人沮喪。在獲得SFA-RB熒光適配器后,她寫道:
"利用NIGHTSEA SFA-RB體視顯微鏡熒光適配器預(yù)篩選是一種很好的方法,可以在我們嘗試在共聚焦上以更高的放大倍率對(duì)肌肉纖維進(jìn)行成像之前快速檢查染色是否成功。
兔腰大肌纖維在白光和熒光下用Alexa Fluor 488 Phalloidin染色。使用NIGHTSEA熒光適配器的白光LED(左)和皇家藍(lán)激發(fā)(SFA-RB)/發(fā)射光+濾光片組制作的圖像。樣本由布朗大學(xué)的Beth Brainerd博士和Natividad Chen博士提供。
另一位研究人員使用斑馬魚作為一個(gè)系統(tǒng)來研究不同的有毒物質(zhì)(藥物,農(nóng)藥,食品添加劑等)如何改變大腦發(fā)育。他寫道:
"在使用NIGHTSEA SFA-RB熒光適配器篩選樣品之前,我必須選擇要安裝的樣品,去共聚焦,然后希望我的一些樣品實(shí)際上是熒光的?,F(xiàn)在我使用NIGHTSEA體視顯微鏡熒光適配器來預(yù)篩選我的樣品,通過確保我成像的樣品是熒光的,我節(jié)省了時(shí)間和金錢。
轉(zhuǎn)基因斑馬魚大腦共聚焦圖像
轉(zhuǎn)基因斑馬魚(Dania rerio)大腦的共聚焦圖像。Kaede蛋白 – 綠色是未轉(zhuǎn)換的,紅色是光轉(zhuǎn)換的。圖片由布朗大學(xué)Creton Lab的Robert Thorn提供。
英文原文:
Pre-Screening Samples for Fluorescence
The NIGHTSEA Model SFA Stereo Microscope Fluorescence Adapter can turn your routine laboratory stereo microscope into a valuable tool for pre-screening your sample preparations for fluorescence before moving on to higher resolution systems.
The Challenge
High resolution imaging of biological samples is heavily based on fluorescence techniques. Confocal, 2-photon, and high resolution compound fluorescence microscopes are almost always a limited resource. They are often located only in imaging core facilities and accessible on a scheduled, pay-per-use basis.
The processes for introducing fluorophores to specimens are not always successful. Staining, introduction of GFP-bearing plasmids to cells, immunohistochemistry – all are fallible. It is not unusual to spend time searching for fluorescence on a high end system when there is not even any there to be found.
The Practical Solution
The NIGHTSEA SFA enables fluorescence pre-screening of specimens on a standard stereo microscope. The detail that you see is not important – the simple presence or absence and general location of fluorescence lets you know whether it is worth taking your specimen to the imaging core. Between the direct expense of the use fee and the time wasted to look at a non-fluorescent specimen it will not take many saved trips for the NIGHTSEA system to more than pay for itself.
One researcher’s work requires staining rabbit psoas muscle fibers with Alexa Fluor 488 Phalloidin. There was some frustration with samples that did not take up the stain. After acquiring the SFA she wrote:
“The NIGHTSEA fluorescence setup is a great way to quickly check whether the stain was successful before we try to image the muscle fiber at a higher magnification on the confocal.”
Rabbit psoas muscle fibers stained with Alexa Fluor 488 Phalloidin
Rabbit psoas muscle fibers stained with Alexa Fluor 488 Phalloidin, in white light and fluorescence. Images made using NIGHTSEA’s white LED (left) and the Royal Blue excitation/emission light+filter set. Samples courtesy of Dr. Beth Brainerd and Natividad Chen, Brown University.
Another researcher uses zebrafish as a system to look at the way different toxicants (pharmaceuticals, pesticides, food additives, etc.) alter brain development. He writes:
“Before using NIGHTSEA to screen my samples, I would have to select samples to mount, go to the confocal and then hope that some of my samples were actually fluorescent. Now that I use NIGHTSEA to prescreen my samples I save both time and money by making sure the only samples I image are fluorescent.”
Confocal image of brain of transgenic zebrafish
Confocal image of brain of transgenic zebrafish (Dania rerio). Kaede protein – green is unconverted, red is photoconverted. Image courtesy of Robert Thorn, Creton Lab, Brown University.