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Fulfills a wide range of application requirements
The iBright Imaging Systems offer up to five imaging modes to support your multiple application requirements. Efficiently and easily capture data from protein gels, nucleic acid gels, chemiluminescent western blots, fluorescent western blots, and more.
| Image mode | What kind of signal can be captured? |
| Protein gel |
Colorimetric staining of gels (e.g., Coomassie, silver) and membranes (e.g., Ponceau S, Thermo Scientific Pierce Reversible Protein Stain), fluorescent staining of gels (e.g., Invitrogen SYPRO Ruby stain) |
| Nucleic acid gel |
Ethidium bromide and Invitrogen SYBR stains |
| Chemiluminescent blot |
Chemiluminescence using all popular HRP and AP substrates (e.g., Thermo Scientific SuperSignal and Invitrogen WesternBreeze substrates) |
| Fluorescent blot |
Fluorescence with popular RGB (visible range) and near-IR fluorophores (e.g., Invitrogen Alexa Fluor and Alexa Fluor Plus conjugates) |
| Universal |
Custom mode to image samples containing multiple signals, such as chemiluminescence, fluorescence, colorimetric stains, and/or visible images; image display is similar to fluorescent blot mode and allows one to assign false color to any sample |
Core imaging applications
Specialty plate-based imaging applications
Qualitative visible imaging applications*
Images pictured for fluorescent western blots, stained nucleic acid gels, colorimetric stained protein gels, GFP expressing colonies, visible stained colonies, and leaf sections are shown in pseudocolor (false color applied). Data is captured in grayscale.* Note that qualitative visible imaging applications provide a qualitative visualization of the object or confirmation of the presence of signal and are not recommended for quantitation.
Optimized E-Gel precast agarose gel imaging and analysis
Leveraging the iBright Tray Adapters for E-Gel Agarose Gels helps streamline the image capture and analysis of our 11, 22, 48, and 96-well E-Gel configurations through appropriately centering the E-gels upon the iBright transilluminator and by blocking the E-gel label or barcode from contributing non-specific fluorescence.
Furthermore, our deconvolution workflow greatly simplifies the interpretation of high-throughput 96-well E-gel results because deconvolution makes it easier to review the data from lane-to-lane to better understand the results.
Touchscreen interface
The 12.1-inch LCD touchscreen interface has a simple, logical layout of functions and features, making our systems easy to use with minimal training. Workflows are similar between imaging modes, delivering a smooth imaging experience regardless of sample type.
Powerful camera technology
The 9.1-megapixel cooled CCD camera captures crisp, clear, publication-quality images. High resolution enables more binning (pixel combining) options which provides flexibility for adjusting resolution and sensitivity based on need.
Smart Exposure technology rapidly determines optimal exposure time, minimizing the potential for over or underexposed images and the need to repeat exposures to get the desired signal.
For chemiluminescent western blot samples with widely varying expression levels, Smart Range HDR (high dynamic range) technology can help maximize the linear dynamic range. This feature leverages two different exposures of the same sample, a short exposure for capturing medium-to-high abundant proteins and a long exposure for capturing low abundant proteins. After capture, the two different images will be combined into a single 16-bit HDR image that contains both the medium-to-high and low abundant signal intensities to effectively extend the linear dynamic range beyond what is achievable with a single short or single long exposure time.
Automation streamlines operation
The iBright 1500 Series Imaging Systems automatically determine the sample position and can rotate samples left or right up to 10° on a mechanically rotating sample stage. This automation eliminates the need for repeated opening of the sample drawer to reposition your sample to achieve proper alignment. In addition, mechanical rotation eliminates the need to digitally rotate the sample, which preserves the integrity of the data.
Digital rotation vs. mechanical rotation. (A) Pixels rotate with digital rotation so bands appear jagged. With mechanical rotation, the sample itself rotates, so bands remain smooth in appearance as the pixels remain aligned. (B) Graphic depicting iBright Imaging System sample stage before and after rotation.
In addition, iBright Imaging Systems automatically adjust the focus for each level of zoom, to maximally utilize the 22.5 cm x 18.0 cm field of view. If imaging a single blot, the camera will automatically zoom up to 2X zoom (1-2X zoom is mechanical zoom with iBright 1500 Series Imaging Systems, 1-2X zoom is digital zoom with the iBright CL750 Imaging system). Mechanical zoom maximizes sensitivity by moving the camera closer to the sample stage and thus reduces focal length. The iBright 1500 Series Imaging Systems also provide additional 1-4X digital zoom for a combined zoom level of 1-8X.
Zoom function. (A) Unzoomed image of a fluorescent western blot. (B) Blot at 2X zoom. (C) Blot at 4X zoom. (D) Blot at 8X zoom. (blot not repositioned during successive zooms)
Large field of view in a small footprint
The large field of view (22.5 x 18.0 cm) enables capture of up to 4 mini blots or gels.
Accelerate your work with fluorescent multiplexed western blots
Expand the possibilities—The iBright FL1500 model features five fluorescence channels, permitting up to 4-color fluorescent western blot multiplexing and expanding your possibilities for studying multiple proteins in a single blot. Obtain meaningful and representative comparisons to enhance your experiments. Smart Exposure technology further improves acquisition of multiplex fluorescent western blot data by ensuring that signal-to-noise ratios are optimized for each fluorescent channel separately.
Filter sets pre-installed in iBright FL1500 Imaging Systems for visible light range (RGB) and near infrared range (NIR) fluorescent western blotting applications are outlined in the table
Excitation channel |
Filter range (nm) |
Emission channel |
Filter range (nm) |
Example compatible fluorophores |
EX1 |
455-485 |
EM1 |
508-557 |
Alexa Fluor Plus 488, Alexa Fluor 488 |
EX2 |
515-545 |
EM2 |
568-617 |
Alexa Fluor Plus 555, Alexa Fluor 546 |
EX3 |
608-632 |
EM3 |
675-720 |
Alexa Fluor Plus 647, Alexa Fluor 594 |
EX4 |
610-660 |
EM4 |
710-730 |
Alexa Fluor Plus 680, Alexa Fluor 680 |
EX5 |
745-765 |
EM5 |
800-850 |
Alexa Fluor Plus 800, Alexa Fluor 790 |
Green-LED transilluminator
The iBright Imaging Systems utilize a transilluminator based on green LEDs, which effectively excite popular DNA dyes such ethidium bromide and SYBR Green dye and offers several additional benefits.
No harmful UV rays: While UV light effectively excites many fluorescent dyes and stains, UV light is a health hazard. Further, prolonged exposure to UV light can damage DNA samples, and compromise the integrity of samples to be used for downstream applications, such as subcloning.
No mercury waste: UV transilluminator bulbs may contain mercury, a hazardous substance, and therefore require special care for handling and disposal.
Longer lifetime: LED bulbs have a substantially longer real-time life than fluorescent UV bulbs, which can add up to considerable cost savings over the lifetime of the instrument.
Protein normalization workflow
To account for variability in samples due to unequal protein sample concentration, inconsistent sample loading onto the gel, and uneven transfer of protein from the gel to membrane, an extra step called normalization is often performed by comparing proteins of interest to internal housekeeping protein controls such as GAPDH, β-tubulin, β-actin, and more. Although housekeeping proteins have been the historically popular choice for normalization, newer strategies involving the comparison of the protein of interest to total lane protein have emerged as a potentially more accurate alternative.
To support the methods best suited for your experiment, iBright Imaging Systems and iBright Analysis Software provide multiple quantitation and normalization options to monitor or mathematically compensate for experimental or sample variability.
Total protein normalization using the No-Stain Protein Labeling Reagent. Invitrogen™ Bolt™ 4-12% Bis-Tris Plus gels were loaded with HeLa lysate ranging from 10 to 50 µg and electrophoresed using MES running buffer. Proteins from the gels were transferred onto mini PVDF membranes using the Invitrogen™ iBlot™ 2 Gel Transfer Device with iBlot 2 Transfer Stacks (P0 protocol for 7 minutes). The PVDF membranes were washed twice for 2 min with 20 mL of ultrapure water on a rotating platform, whereupon they were labeled with 10 mL of a working solution of No-Stain Protein Labeling Reagent on a rotating platform for 10 minutes. The membranes were then washed 3 times for 2 minutes with 20 mL of ultrapure water on a rotating platform, followed by addition of Invitrogen™ primary antibodies against β-actin (Cat. No. AM4302), GAPDH (Cat. No. 398600), and α-tubulin (Cat. No. 138000), and Invitrogen™ Goat Anti–Mouse IgG Alexa Fluor™ Plus 680 secondary antibody (Cat. No. A21058). The blot was imaged using the Invitrogen™ iBright™ FL1500 Imaging System. The iBright software was used to quantitate the total protein signal in the lanes. The R² value of the plotted data for the entire range of total protein signal was determined to be 0.9990, whereas the R² values for β-actin, GAPDH, and α-tubulin were 0.8851, 0.9438, and 0.8332, respectively.