轻触指尖,定格明亮印记

12.1寸全触屏一体机,轻松、高效捕捉和分析Western blot印迹膜/ 凝胶成像

910万像素冷CCD,拍摄图片更清晰;
一次可捕捉4块小型印迹膜或者凝胶,处理通量更高;
一张印迹膜上可同时拍摄4种荧光(包括RGB和近红外),检测样本量更多;
具有64G存储空间以及10G icloud云平台,可随时随地获取和分析数据;
拍摄高品质Western blot图像,您无需成为专家。

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使用Invitrogen iBright智能成像系统,对您的免疫印迹和凝胶轻松、高效地进行成像和数据分析。这些高性能仪器具有先进的自动化功能和易于使用的界面,适用于所有经验水平的研究人员,提升了免疫印迹体验。即刻体验iBright成像系统,在全新蛋白质免疫印迹世界中锐意前行,迎接挑战。

观看视频:Invitrogen iBright智能成像系统——轻触指尖,快速获取明亮Western blot印迹

观看视频:如何通过iBright智能成像系统拍摄图片

观看视频:如何从iBright智能成像系统中导出图片

观看视频:如何使用iBright智能成像系统分析图片

“iBright成像仪的用户体验非常友好!”

– Rebecca Sinnott DeVaux, SUNY

展开以下选项卡,了解我们的iBright智能成像系统如何帮助您将免疫印迹和凝胶成像流程变的更流畅。

操作自如—电容式LCD触摸屏的操作方式与其他常用的触摸屏设备类似,您可轻松开启体验之旅。

Figure 1. Touch screen controls.

放置样本后即刻运行—使用电动抽屉放入样品。用户界面采用简单、合理的功能与特性布局,使iBright智能成像系统易于使用,几乎不需要培训。不同成像模式的工作流程相似——不论哪种样本类型均可获得流畅的成像体验。 

简单的样品对齐、聚焦和缩放———iBright智能成像系统可自动确定样本位置,并通过机械式旋转载物台向左或向右旋转样本最多10°,因此无需打开样本抽屉、反复摆放样本即可正确对齐。机械式旋转的效果优于数字式旋转,因为数字式旋转可导致数据的更改。 

iBright智能成像系统可自动确定样品是否需要缩放,从而最大限度地利用视野。如果对单一印迹成像,相机将向样品移动(最多2x缩放)。此操作将使相机更靠近样品,缩短光线照射到相机传感器的距离,从而进一步提高灵敏度。iBright成像系统可自动调整每个缩放级别的对焦,获取清晰的图像。 

Figure 3. Sample rotation. Graphic depicting iBright Imaging System stage before and after rotation.
Fig. 4. Digital vs mechanical sample rotation. Left: digital rotation (10°)—pixels rotate with digital rotation, so bands appear jagged. Right: Mechanical rotation (10°)—sample rotates so bands remain smooth as pixels remain aligned
Figure 5. Zoom function. At 1x zoom, the field of view is 22.5 cm (W) x 18.0 cm (H). Four full-sized mini blots or gels can be accommodated (left panel). The same blot outlined in the left panel, when repositioned in the center of the field of view, at 2x zoom (right panel).

可在短时间内完成分析—iBright智能成像系统采用自动化机载数据分析,可实现瞬时的泳道和条带鉴定以及分子量标准的叠加,极大地简化了基础的成像后数据分析。支持直接在仪器上进行定量和光密度分析。能够同时分析多达4张印迹膜或凝胶,大大提高了通量。

Figure 6. Automatic lane and band identification of four mini blots in chemiluminescent blot acquisition mode.

采集清晰的图像—智能成像技术可快速确定最佳曝光时间,有助于将图像过度曝光或曝光不足的可能性降至最低,无需重复曝光即可获得理想的信号。iBright智能成像系统结合了智能成像技术与灵敏的910万像素(MP)冷CCD相机,具有强大的成像功能,有助于检测蛋白质表达的微小差异。

Figure 7. Detect subtle protein modifications using iBright Imaging Systems. HeLa cells were stimulated with IFN-alpha to induce protein phosphorylation, or with staurosporine to induce protein cleavage. Following stimulation, cells were lysed with IP Lysis Buffer (Cat. No. 87787). Equal amounts (20 µg) of the cell lysates were loaded onto 4–20% Tris-glycine gels (Cat. No. WT4202BOX). Proteins were transferred to nitrocellulose membranes (Cat. No. 88018) using the Thermo Scientific Pierce Power Blotter (Cat. No. 22834), and the membranes were then blocked with 5% milk in TBS-Tween 20 (TBST, Cat. No. 28360) for at least 30 min. The membranes were probed overnight at 4°C with antibodies against PARP (Cat. No. MA5-15031, 1:1,000 dilution) (left panel), or phospho-STAT3 (Cat. No. MA5-15193, 1:500 dilution) and STAT3 (Cat. No. MA1-13042, 1:5,000 dilution) (right panel). Blots were washed 3 times for at least 10 min each in TBST, and probed with HRP-conjugated goat anti-rabbit secondary antibody (Cat. No. 31460, 20 ng/mL) (left panel) or HRP-conjugated goat anti-mouse secondary antibody (Cat. No. 31430, 20 ng/mL) (right panel) for 1 hr at room temperature. Blots were again washed 3 times for at least 10 min each in TBST, and developed using SuperSignal West Dura substrate (Cat. No. 34076). Bands were visualized on the iBright FL1000 Imaging System with 2 min exposure (PARP), 3 min exposure (phospho-STAT3) and 52 sec exposure (STAT3).

Figure 8. iBright Imaging Systems feature a powerful 9.1 MP camera for greater sensitivity compared to instruments with a lower-resolution camera. Two-fold serial dilutions of HeLa cell lysate (starting at 80 µg/lane) were loaded onto 4–20% Tris-glycine gels (Cat. No. WT4202BOX). Proteins were transferred to nitrocellulose membranes (Cat. No. 88018) using the Pierce Power Blotter (Cat. No. 22834), and the membranes were then blocked with 5% milk in TBS-Tween 20 (TBST, Cat. No. 28360) for at least 30 min. The membranes were probed overnight at 4°C with antibodies against Ku80 (Cat. No. MA5-14953, 1:1,000 dilution) or DDX3 (Cat. No. PA5-17165, 1:1,000 dilution). Blots were washed 3 times for at least 10 min each in TBST, and probed with an HRP-conjugated goat anti-rabbit secondary antibody (Cat. No. 31460, 10 ng/mL (Ku80 blot) and 40 ng/mL (DDX3 blot)) for 1 hr at room temperature. Blots were again washed 3 times for at least 10 min each in TBST, and developed using SuperSignal West Pico PLUS substrate (Cat. No. 34580). Bands were visualized on the iBright FL1000 Imaging System (top blots), another imaging device with a lower-quality CCD camera (bottom blots), each with 10 sec exposures.

Figure 9. Comparison of detection sensitivity and dynamic range for film and the iBright FL1000 Imaging System. A luminometer reference plate emitting light at varying fixed intensities and specific wavelength (540nm) was used to expose film or to acquire an image on the iBright FL1000 Imaging System for 10 sec (left panel). More luminometer spots are visible on the image from the iBright instrument, indicating higher sensitivity compared to film. iBright and film 1 min exposures to the same luminometer plate (middle panel) with graphical analysis. Analysis reveals the better signal linearity and dynamic range of signals acquired using the iBright FL1000 Imaging System compared to film (right panel).

大视野,小体积—iBright智能成像系统具有独特的光路设计,可在相对较小的仪器内获得较大的视野面积。

Figure 10. The large field of view (22.5 x 18.0 cm) enables capture of up to 4 mini blots or gels. Left: four mini blots. Right: signals captured in fluorescent blot mode.

使用寿命更长的反射LED光源—iBright FL1000智能成像系统采用了两个高品质长寿命反射LED光源的简单组合,用于荧光成像应用。宽光谱的白光LED可用作RGB荧光和远红外荧光光源。另一个LED则专为近红外荧光进行了优化。这些光源发出的光线通过配套的激发和发射滤光片,配合多种试剂方案,可以用于蛋白凝胶、核酸凝胶和印迹成像应用(表1)。

表 1.可从凝胶或印迹膜捕获的数据类型概述。

成像能力 可捕获哪种信号?
蛋白质凝胶 凝胶(如考马斯染料、银染试剂)和膜(如Ponceau S、Thermo Scientific MemCode染色剂)的比色染色
核酸凝胶 溴化乙锭和Invitrogen SYBR染料染色
化学发光印迹 使用各种常用HRP和AP底物(如Thermo Scientific SuperSignal和Invitrogen WesternBreeze底物)的化学发光印迹
荧光印迹* 使用常用RGB(可见光范围)和近红外荧光基团(如Invitrogen Alexa Fluor、Alexa Fluor Plus、Thermo Scientific DyLight染料)的荧光染色
*仅FL1000型号。

参见iBright成像仪兼容的染料和试剂列表

加速您的研究—iBright FL1000具有五个荧光通道,可同时对4色荧光蛋白质免疫印迹进行成像,拓展了在单块印迹上同时检测多种蛋白质的能力。从而您可获取更有意义且更具代表性的比较数据,完善您的实验。Smart Exposure智能成像技术优化了每个荧光通道的信噪比,进一步改善了多重荧光免疫印迹数据的获取。

Figure 11. Two-channel (near-IR) imaging of a fluorescent western blot. Several HA-tagged proteins were expressed in HeLa cell extract using the Thermo Scientific 1-Step Human High-Yield Mini IVT Kit (Cat. No. 88890) and appropriate expression-ready clones. The resulting reaction mixtures were prepared for SDS-PAGE and electrophoresed: Erythropoietin precursor, lanes 1-2; Streptokinase, lanes 3-4; Caveolin-1, lanes 5-6; molecular weight marker, lane 7; Argonaute-2, lanes 8-9; Regulatory-associated protein of mTOR, lanes 10-11; Bcl2-associated agonist of cell death, lanes 12-13; Retinoblastoma susceptibility protein, lanes 14-15. The proteins were transferred to a PVDF membrane using the Pierce Power Blotter (Cat. No. 22834), and the membrane was blocked and probed with the following primary antibodies: mouse anti-HA (Cat. No. 26183) and rabbit anti–cyclophilin B (Cat. No. PA1-027A). Cyclophilin-B is present in the HeLa cell extract and serves as a control in this experiment. The membrane was washed and probed with the following secondary antibodies in TBS-Tween 20: goat anti-rabbit Alexa Fluor Plus 800 (Cat. No. A32735) (pseudocolored in red) and goat anti-mouse Alexa Fluor Plus 680 (Cat. No. A32729) (pseudocolored in green). The membrane was washed and imaged on the iBright FL1000 Imaging System.
Figure 12. Four-channel imaging of a fluorescent western blot. Up to four different proteins can be imaged simultaneously on the same blot. HA-tagged RB-1 was expressed in HeLa cell extract using the 1-Step Human High-Yield Mini IVT Kit (Cat. No. 88890) and appropriate expression-ready clones. The resulting reaction mixture was prepared for reducing SDS-PAGE, serially diluted, and electrophoresed on a Novex WedgeWell 4–20% Tris-glycine gel (Cat. No. XP04200PK2). The protein was transferred to a PVDF membrane using the Pierce Power Blotter (Cat. No. 22834), and the membrane was blocked and probed with the following primary antibodies: chicken anti-calreticulin (Cat. No. PA1-903), rabbit anti-HSP90 (Cat. No. PA3-013), and mouse anti-p23 (Cat. No. MA3-414). The membrane was washed and probed with the following secondary antibodies in TBS-Tween 20: goat anti-chicken Alexa Fluor 546 (Cat. No. A11040) (pseudocolored in yellow), goat anti-rabbit Alexa Fluor Plus 800 (Cat. No. A32735) (pseudocolored in green), and goat anti-mouse Alexa Fluor Plus 680 (Cat. No. A32729) (pseudocolored in red). The membrane was again washed and probed for 1 hr with mouse anti-HA primary antibody directly conjugated to Alexa Fluor 488 (Cat. No. 26183-D488) (pseudocolored in blue), in TBS-Tween 20. The membrane was washed and imaged on the iBright FL1000 Imaging System.

iBright智能成像系统利用绿色LED透照仪,可有效激发常用DNA染料,如溴化乙锭和SYBR Green染料,同时具有许多其他优点。

不使用有害的紫外线:尽管紫外光可有效激发许多荧光染料和染色剂,但紫外线会危害健康。此外,长时间暴露于紫外光可能会损伤DNA样品,并影响用于下游应用(如亚克隆)的样品完整性。

不产生含汞废物:紫外透射灯含有有害物质汞,因此需要特殊的操作和处置。

使用寿命更长:LED灯泡的寿命比紫外灯长很多,因此,在仪器的使用寿命内可以节省大量成本。

简单的软件包蕴含强大的分析功能— iBright 分析软件在iBright智能成像系统机载分析特性的基础上,扩展了深度图像调整和数据分析性能。iBright分析软件旨在完善整体直观的成像体验。

Figure 13. iBright Analysis Software features extensive analysis and image adjustment functions.

云连接有助于提高效率—iBright分析软件基于Thermo Fisher Cloud云平台,是Thermo Fisher Connect套装的一部分。数据可以直接从iBright智能成像系统导出并安全地存储在Thermo Fisher Cloud中。由于iBright分析软件是基于网络的,您可以随时随地访问、查看、分析和分享您的数据。此外,使用Thermo Fisher Connect,您还可以确定仪器状态、固件版本和使用历史,加大对投资设备的监管力度。

保障数据安全—Thermo Fisher Connect由Thermo Fisher Cloud提供技术支持。Thermo Fisher Cloud采用强大的安全标准,包括高级别的加密和网络防火墙,从而为您的数据提供安全保障。 Thermo Fisher Cloud还提供数据备份和恢复功能,即使在紧急情况下,您的数据仍然安全。 

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