美國(guó)NIGHTSEA專(zhuān)門(mén)致力于研究和開(kāi)發(fā)熒光觀測(cè)裝備。NIGHTSEA轉(zhuǎn)基因生物觀測(cè)利器包括:SFA-RB體視顯微鏡熒光適配器,DFP-1雙熒光蛋白手電筒系列,BLS2單熒光蛋白手電筒系列等。
(一)美國(guó)NIGHTSEA轉(zhuǎn)基因生物觀測(cè)利器的應(yīng)用之——轉(zhuǎn)基因生物觀測(cè):如小鼠幼崽,果蠅幼蟲(chóng),線蟲(chóng),斑馬魚(yú),等等。
1. 美國(guó)NIGHTSEA轉(zhuǎn)基因生物觀測(cè)利器—觀測(cè)轉(zhuǎn)基因果蠅
2. 美國(guó)NIGHTSEA轉(zhuǎn)基因生物觀測(cè)利器—觀測(cè)轉(zhuǎn)基因斑馬魚(yú)
3. 美國(guó)NIGHTSEA轉(zhuǎn)基因生物觀測(cè)利器—觀測(cè)轉(zhuǎn)基因爪蟾
4. 美國(guó)NIGHTSEA轉(zhuǎn)基因生物觀測(cè)利器—觀測(cè)轉(zhuǎn)基因線蟲(chóng)
5.美國(guó)NIGHTSEA轉(zhuǎn)基因生物觀測(cè)利器—觀測(cè)轉(zhuǎn)基因蠑螈
6.美國(guó)NIGHTSEA轉(zhuǎn)基因生物觀測(cè)利器—觀測(cè)轉(zhuǎn)基因擬南芥種子
(二)美國(guó)NIGHTSEA轉(zhuǎn)基因生物觀測(cè)利器的應(yīng)用之——樣本預(yù)篩選和輔助解剖
熒光是細(xì)胞生物學(xué)、神經(jīng)生物學(xué)等多個(gè)研究領(lǐng)域中廣泛使用的工具。nightsea的DFP-1, BLS2和SFA-RB可以用于樣本的預(yù)篩選、輔助解剖等,下面列舉幾個(gè)實(shí)例:
1. 熒光蛋白標(biāo)記的工程病毒載體在神經(jīng)元功能研究中應(yīng)用十分廣泛,常通過(guò)顱內(nèi)注射研究特定腦區(qū)的功能。在此中,常需要對(duì)病毒表達(dá)區(qū)域進(jìn)行快速解剖用于特定的生化分析。為了提高快速解剖定位度,研究人員選用nightsea的BLS2 Bluestar手電筒觀測(cè)系列,通過(guò)配合nightsea濾光眼鏡準(zhǔn)確定位注射表達(dá)了熒光蛋白的伏核區(qū)(文獻(xiàn)– Xuan Li and Marina E. Wolf, 2011. Visualization of virus-infected brain regions using a GFP-illuminating flashlight enables accurate and rapid dissection for biochemical analysis. J. of Neuroscience Methods, Vol. 201, Issue 1, pp. 177-179. )
EGFP標(biāo)記的慢病毒載體注射進(jìn)小鼠大腦區(qū)域(Marina Wolf, Rosalind Franklin University)
2. 研究者從小鼠大腦中提取GFP標(biāo)記的背紋體。這項(xiàng)困難的工作因有了NIGHTSEA的DFP-1或BLS2(手電筒和眼鏡組合)而變得簡(jiǎn)單:很容易看到大腦中的目標(biāo)區(qū)域,讓解剖更準(zhǔn)確、。
小鼠大腦中GFP標(biāo)記的背紋體(Charles Mazel. Sample photographed at laboratory of Stefano Vicini, Georgetown University)
3. 用DFP-1雙熒光蛋白手電筒,BLS2單熒光蛋白手電筒,或SFA-RB體視顯微鏡熒光適配器,可以快速檢測(cè)樣本是否成功染色(Alexa Fluor 488 Phalloidin標(biāo)記),如下圖:
兔腰肌的肌纖維(用Alexa Fluor 488 Phalloidin標(biāo)記)
左圖:白光觀測(cè) 右圖:Royal Blue熒光觀測(cè)(Motic SMZ-186體視顯微鏡觀測(cè))
4.用SFA-RB體視顯微鏡熒光適配器以斑馬魚(yú)為模型,觀測(cè)不同有害化學(xué)物(醫(yī)藥、農(nóng)藥、食品添加劑等)對(duì)大腦發(fā)育的影響。觀測(cè)圖片如下:
基因斑馬魚(yú)大腦共聚焦圖像
圖片由Robert Thorn(Creton Lab, 布朗大學(xué))提供
5. 熒光是珊瑚采集研究的重要工具?,F(xiàn)在,可以用經(jīng)濟(jì)又實(shí)惠的SFA-RB體視顯微鏡熒光適配器安裝到體視顯微鏡上檢測(cè)珊瑚。
下面的圖片是通過(guò)立體顯微鏡觀看珊瑚蟲(chóng),由UNCW威爾明頓納大學(xué)Alina Szmant 博士與NIGHTSEA Charles Mazel博士提供
6. 用于結(jié)核分枝桿菌重組株的篩選
在結(jié)核分枝桿菌中成功構(gòu)建了同源重組系統(tǒng),利用該系統(tǒng)構(gòu)建了rv1364c、pstP跨膜區(qū)、pstP胞外區(qū)三個(gè)突變株,得到雙交換突變株的效率為25%-62.5%,從雙交換突變株得到無(wú)痕缺失突變株的效率為100%。通過(guò)gfp作為熒光標(biāo)記基因,利用NightSea BlueStar藍(lán)光手電筒和濾光眼鏡,可以對(duì)平板上的基因缺失株菌落直接進(jìn)行快速判定。
采用nightsea BLS2 BlueStar 藍(lán)色手電筒和濾光鏡對(duì)平板上的結(jié)核分枝桿菌綠色熒光蛋白菌落進(jìn)行快速鑒定。該方法可以從幾百個(gè)菌落中瞬間鎖定陽(yáng)性克隆。平板上的菌落照片拍攝用單反相機(jī)加載購(gòu)自nightsea公司的濾光片實(shí)現(xiàn)。
圖片源自[結(jié)核分枝桿菌同源重組基因敲除系統(tǒng)的構(gòu)建和應(yīng)用]
熒光是細(xì)胞生物學(xué)、神經(jīng)生物學(xué)等領(lǐng)域中廣泛使用的工具。美國(guó)nightsea致力于研究和開(kāi)發(fā)熒光觀測(cè)裝備,其產(chǎn)品系列有DFP-1雙熒光蛋白手電筒系列,BLS2單熒光蛋白手電筒系列,SFA-RB體視顯微鏡熒光適配器等。
DFP-1 Dual Fluorescent Protein Flashlight 雙熒光蛋白手電筒系列
DFP-1雙熒光蛋白手電筒,有兩個(gè)可轉(zhuǎn)換的高強(qiáng)度LED,可激發(fā)高強(qiáng)度光源。目前,綠色熒光(GFP、EGFP、fluorescein等)和紅色熒光(DsREed,TdTomato等)是常用的類(lèi)型,nightsea常規(guī)的DFP-1雙熒光蛋白手電筒是激發(fā)藍(lán)光和綠光,再分別使用黃色濾光鏡和紅色濾光鏡,就可以分別檢測(cè)綠色熒光蛋白(GFP、EGFP、fluorescein等)和紅色熒光蛋白(DsREed,TdTomato等)了。
Good News
標(biāo)準(zhǔn)的DFP-1雙熒光蛋白觀測(cè)鏡,只有RB-Royal Blue和GR-Green兩種熒光激發(fā)裝置,也就是說(shuō)標(biāo)準(zhǔn)的DFP-1只能用于觀測(cè)綠色熒光蛋白和紅色熒光蛋白。
現(xiàn)在,您可以選擇以下任何兩種熒光裝置,組裝成自己專(zhuān)屬的雙熒光蛋白手電筒。
熒光裝置型號(hào) | Excitation 激發(fā)光 | Emission 發(fā)射光 | 可觀測(cè)熒光 |
RB-Royal Blue | 440-460nm | 500nm LP | GFP, eGFP, fluorescein.... |
CY-Cyan | 490-515nm | 550nm LP | YFP, Venus, Lucifer Yellow... |
GR-Green | 510-540nm | 600nm LP | DsRed, dTomato... |
VI-Violet | 400-415nm | 460nm LP | CFP, BFP... |
UV-Ultra Violet | 360-380nm | 415nm LP | DAPI |
注:
1.RB-Royal Blue用于觀測(cè)綠色熒光蛋白
GFP:綠色熒光;EGFP:增強(qiáng)型綠色熒光;fluorescein熒光素
2.CY-Cyan用于觀測(cè)黃色熒光蛋白
YFP:黃色熒光蛋白;Venus:金色;Lucifer Yellow:熒光黃
3.GR-Green用于觀測(cè)紅色熒光蛋白
DsRed:紅色熒光蛋白;dTomato:番茄紅
4.VI-Violet用于觀測(cè)紫色熒光蛋白
CFP:青色熒光蛋白;BFP:堿性胎兒蛋白
5.UV-Ultra Violet用于觀測(cè)紫外光
DAPI:藍(lán)色熒光
nightsea 2018年 產(chǎn)品列表:
貨號(hào) | 品名 | 規(guī)格 | 價(jià)格 | 品牌 |
SFA-TENT | Eclipse MicroTent | each | 5525 | nightsea |
SFA-EYE-S | Tru-Block Eye Shields - Standard | set | 306 | nightsea |
SFA-EYE-C | Tru-Block Eye Shields - Compact | set | 306 | nightsea |
SFA-BLH-RB | Light Head-Royal Blue | each | 4250 | nightsea |
SFA-BLH-CY | Light Head-Cyan | each | 4250 | nightsea |
SFA-BLH-GR | Light Head-Green | each | 4250 | nightsea |
SFA-BLH-VI | Light Head-Violet | each | 4250 | nightsea |
SFA-BLH-UV | Light Head-Ultraviolet | each | 6545 | nightsea |
SFA-BF-RB | NIGHTSEA Barrier Filter-Royal Blue | each | 3536 | nightsea |
SFA-BF-GO | NIGHTSEA Barrier Filter-Green Only | each | 3536 | nightsea |
SFA-BF-CY | NIGHTSEA Barrier Filter-Cyan | each | 3536 | nightsea |
SFA-BF-GR | NIGHTSEA Barrier Filter-Green | each | 3536 | nightsea |
SFA-BF-VI | NIGHTSEA Barrier Filter-Violet | each | 3536 | nightsea |
SFA-BF-UV | NIGHTSEA Barrier Filter-Ultraviolet | each | 3536 | nightsea |
SFA-SH-RB | NIGHTSEA Shield-Royal Blue | each | 1785 | nightsea |
SFA-SH-GO | NIGHTSEA Shield-Green Only | each | 1785 | nightsea |
SFA-SH-CY | NIGHTSEA Shield-Cyan | each | 1785 | nightsea |
SFA-SH-GR | NIGHTSEA Shield-Green | each | 1785 | nightsea |
SFA-SH-VI | NIGHTSEA Shield-Violet | each | 1785 | nightsea |
SFA-SH-UV | NIGHTSEA Shield-Ultraviolet | each | 1785 | nightsea |
SFA-AD | NIGHTSEA Adapter | each | 2856 | nightsea |
SFAZ-AD | Leica EZ4 Adapter only | each | 2856 | nightsea |
SFA-XL-AD | NIGHTSEA Oversize Adapter | each | 2856 | nightsea |
SFA-DIM-N | NIGHTSEA DIM Base | each | 11305 | nightsea |
SFA-PULSE-N | NIGHTSEA PULSE Base | each | 11305 | nightsea |
SFA-BASE | NIGHTSEA Standard Base | each | 7055 | nightsea |
SFA-LHH-L | Light Head Hanger, long working distance | each | 1615 | nightsea |
SFA-LHH-S | Light Head Hanger, short working distance | each | 1615 | nightsea |
SFA-LHC | Single Light Head Cable, V1 | each | 510 | nightsea |
SFA-LHC-BNC | Single Light Head Cable, BNC | each | 595 | nightsea |
SFA-DLHC | Dual Light Head Cable, V1 | each | 1105 | nightsea |
SFA-DLHC-BNC | Dual Light Head Cable, BNC | each | 1105 | nightsea |
SFA-HK-L | Single hanger kit, long working distance, V1: SFA-LHC + SFA-LHH-L | kit | 2125 | nightsea |
SFA-HK-S | Single hanger kit, short working distance, V1: SFA-LHC + SFA-LHH-S | kit | 2125 | nightsea |
SFA-HK-L-BNC | Single hanger kit, long working distance, BNC: SFA-LHC-BNC + SFA-LHH-L | kit | 2125 | nightsea |
SFA-HK-S-BNC | Single hanger kit, short working distance, BNC: SFA-LHC-BNC + SFA-LHH-S | kit | 2125 | nightsea |
SFA-DHK-L | Dual hanger kit, long working distance, V1: SFA-DLHC + 2x SFA-LHH-L | kit | 4165 | nightsea |
SFA-DHK-S | Dual hanger kit, short working distance, V1: SFA-DLHC + 2x SFA-LHH-S | kit | 4165 | nightsea |
SFA-DHK-L-BNC | Dual hanger kit, long working distance, BNC: SFA-DLHC-BNC + 2x SFA-LHH-L | kit | 4165 | nightsea |
SFA-DHK-S-BNC | Dual hanger kit, short working distance, BNC: SFA-DLHC-BNC + 2x SFA-LHH-S | kit | 4165 | nightsea |
SFA-SWK-S-BNC | Switch Box Kit, short working distance: BNC switch box, 3 BNC cables, one with female/female gooseneck adapter, 2 LH Hangers | kit | 5950 | nightsea |
SFA-SWK-L-BNC | Switch Box Kit, long working distance: BNC switch box, 3 BNC cables, one with female/female gooseneck adapter, 2 LH Hangers | kit | 5950 | nightsea |
DFP-1 | DFP Flashlight with RB and GR LEDs, matching filter glasses, carrying case | each | 12835 | nightsea |
DFP-CG | DFP Flashlight with CY and GR LEDs, matching filter glasses, carrying case | each | 13855 | nightsea |
DFP-RC | DFP Flashlight with RB and CY LEDs, matching filter glasses, carrying case | each | 13855 | nightsea |
DFP-VC | DFP Flashlight with VI and CY LEDs, matching filter glasses, carrying case | each | 13855 | nightsea |
DFP-VG | DFP Flashlight with VI and GR LEDs, matching filter glasses, carrying case | each | 13855 | nightsea |
DFP-VR | DFP Flashlight with VI and RB LEDs, matching filter glasses, carrying case | each | 13855 | nightsea |
DFP-UC | DFP Flashlight with UV and CY LEDs, matching filter glasses, carrying case | each | 16065 | nightsea |
DFP-UG | DFP Flashlight with UV and GR LEDs, matching filter glasses, carrying case | each | 16065 | nightsea |
DFP-UR | DFP Flashlight with UV and RB LEDs, matching filter glasses, carrying case | each | 16065 | nightsea |
DFP-UV | DFP Flashlight with UV and VI LEDs, matching filter glasses, carrying case | each | 16065 | nightsea |
BLS-1 | BlueStar light plus style VG1 filter glasses | each | 3485 | nightsea |
BLS-2 | BlueStar light plus style VG2 filter glasses | each | 3655 | nightsea |
BLS-3 | BlueStar light plus style VG3 filter glasses | each | 3655 | nightsea |
VG-1 | Yellow filter glasses, style 1 | each | 578 | nightsea |
VG-2 | Yellow filter glasses, style 2 | each | 935 | nightsea |
VG-3 | Yellow filter glasses, style 3 | each | 935 | nightsea |
RG-2 | Red filter glasses, style 2 | each | 935 | nightsea |
FG-UV | Filter glasses, Ultraviolet | each | 935 | nightsea |
FG-VI | Filter glasses, Violet | each | 935 | nightsea |
FG-GO | Filter glasses, Green Only | each | 935 | nightsea |
FG-CY | Filter glasses, Cyan | each | 935 | nightsea |
SFA-PULSE-R | PULSE Option - Retrofit | each | 4675 | nightsea |
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