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Explore the imaging, analysis, and automation capabilities available with iBright imaging systems. Learn how integrated imaging technologies, automated image acquisition, quantitative analysis tools, and compliance-ready workflows help simplify western blot imaging, gel documentation, and protein analysis.
Explore the western blot workflow
Features that simplify western blot imaging and analysis
iBright imaging systems combine advanced imaging technologies, automated image acquisition, and integrated analysis tools in a single platform. Explore the imaging, analysis, automation, and compliance capabilities that enable researchers to capture, quantify, and document protein analysis experiments with confidence.
What applications can be imaged with iBright imaging systems?
iBright imaging systems support western blot imaging, protein and nucleic acid gel documentation, chemiluminescent detection, fluorescent imaging, colorimetric detection, and multiplex imaging on a single platform. Select imaging modes based on sample type, detection chemistry, and analysis requirements.
Image application |
Supported sample types and detection modes |
Protein gel |
Image colorimetric- and fluorescently stained protein gels, including Coomassie, silver, Ponceau S, Reversible Protein Stain, and SYPRO Ruby. |
Nucleic acid gel |
Image DNA and RNA gels stained with ethidium bromide, SYBR dyes, and other common nucleic acid stains. |
Chemiluminescent blot |
Image chemiluminescent western blots using HRP- or AP-based detection substrates. |
Fluorescent blot |
Image and quantify fluorescent western blots using visible and near-infrared fluorophores, including multiplex protein detection. |
Universal |
Capture and analyze multiple signal types—including chemiluminescent, fluorescent, colorimetric, and visible samples—in a single imaging mode. |
Depending on the system configuration, iBright imaging systems support protein and nucleic acid gel imaging, chemiluminescent and fluorescent western blot imaging, colorimetric detection, and multi-signal imaging workflows.
The examples below demonstrate how these imaging modes can be applied across western blotting, gel documentation, colony imaging, and specialty imaging workflows.
Core imaging applications
Capture and analyze western blots, protein gels, and nucleic acid gels using chemiluminescent, fluorescent, and colorimetric detection methods.
Fluorescent western blots
Chemiluminescent western blots
Combined fluorescent and chemiluminescent western blots
Colorimetric western blots
Fluorescent stained nucleic acid gels
Fluorescent stained protein gels
Colorimetric stained protein gels
Colorimetric membrane stains
Specialty plate-based imaging applications
Expand imaging capabilities beyond western blotting to include colony counting and In-Cell Western assays.
Fluorescent colonies (e.g. GFP expression)
Visible colonies (e.g. crystal violet stained)
In-Cell Western™ image capture and analysis
Related application notes:
Qualitative visible imaging applications*
Visualize visible-light samples such as TLC plates and plant tissues for qualitative assessment and documentation.
TLC-plates
Leaf sections
Note: 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.*
Qualitative visible imaging applications help provide a qualitative visualization of the object or confirmation of the presence of signal and are not recommended for quantitation.
iBright touchscreen interface
The iBright touchscreen interface combines image acquisition, analysis, and data management tools in a single workspace. Researchers can capture images, adjust imaging settings, analyze results, and export data directly from the instrument without switching between software applications.
The touchscreen interface streamlines the process of image capture while still providing users with the ability to control the settings for their experiment.
Interface features
- Sign in: create a new user account or sign in to an existing account
- Gallery: access previously captured images
- Mode selection: dropdown menu to select imaging modes (chemiluminescent blot, fluorescent blot, nucleic acid gel, protein gel, and universal mode)
- Camera lock: maintain camera settings across multiple samples, locks exposure time, zoom, and transillumination settings while disabling mechanical auto-rotation
- Help: on-board help and information
- Settings: access general instrument settings, configurations, and service tools
- Drawer: open or close the sample drawer
- Exposure dial: select or turn the dial to set manual exposure time or fine-tune exposure time
- Capture: acquire images using exposure time set by user or instrument
- Edit channels: open multiple channel options to review and edit each channel
- More options: open image adjustment options
- Live view: go back to live-streaming sample view
- Analyze: initiate analysis workflow using current images
- Export: edit image information and export current images to destinations of choice
- Trash: delete current images
- Split screen: select one-window or two-window image view
- Color option: select color or grayscale image view
- Image view window: display selected image in the window for user view and interaction
The touchscreen interface allows users to acquire images, adjust exposure settings, review results, perform image analysis, and export data directly from the instrument, reducing the need for multiple software platforms and manual workflow steps.
Smart Exposure technology
iBright imaging systems help simplify western blot image acquisition using Smart Exposure and Smart Range HDR technologies that support consistent image quality and reliable protein analysis. These features streamline imaging workflows while helping researchers generate high-quality images for visualization and quantitation.
The 9.1-megapixel cooled CCD camera captures crisp, clear, publication-quality images. High resolution enables more binning (pixel combining) options which help provide 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.
How does Smart Exposure technology work?
Smart Exposure technology automatically determines the optimal exposure time for each image, helping reduce time spent on manual optimization while minimizing the potential for over or underexposed images and the need to repeat exposures.
Comparison of Smart Exposure technology to manually set exposure times. Minimal pixel saturation is observed in the image captured with exposure time determined by Smart Exposure technology, while the range of data captured is maximized. The set of images on the right is the same as the image on the left, but with the saturated pixels feature of iBright imaging systems turned on (saturated pixels are displayed in red). The same blot was imaged using the exposure time determined by Smart Exposure technology, or four manually set exposure times.
What is Smart Range HDR imaging?
Smart Range HDR combines a long and short exposure into a single image, helping researchers visualize both weak and strong protein signals while extending the effective linear dynamic range for western blot quantitation. Smart Range HDR can improve visualization of proteins with widely varying expression levels and reduce the need for repeated image acquisitions at different exposure settings.
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.
SmartRange HDR improved the detection limit of p23 4-fold compared to the Smart Exposure feature. HeLa lysate was serially diluted 1:2 in sample buffer (20 µg, 10 µg, …10 ng), prepared for SDS-PAGE and electrophoresed on a Novex WedgeWell 4–20% Tris-Glycine gel. The protein was transferred to nitrocellulose membrane and probed for p23. The resulting western blot was imaged on the iBright imager using Smart Exposure and SmartRange HDR.
Related application note:
Difference between Smart Exposure and Smart Range HDR
Technology |
Purpose |
Smart Exposure |
Automatically identifies the optimal exposure time for image acquisition. |
Smart Range HDR |
Combines two exposures to capture both weak and strong signals in a single image. |
Simplify sample positioning and image acquisition
iBright imaging systems automate sample positioning, alignment, focus, and zoom adjustments to help reduce manual setup and improve image consistency. Automated rotation and zoom functions help optimize image quality while preserving sample integrity and reducing workflow variability.
Why use mechanical rotation instead of digital rotation?
Mechanical rotation physically repositions the sample rather than digitally manipulating the image. This helps preserve image quality, maintain smooth band appearance, and reduce artifacts that can occur when images are digitally rotated.
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 reduces the need for repeated opening of the sample drawer to reposition your sample to achieve proper alignment. In addition, mechanical rotation reduces 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.
How does automated zoom improve imaging?
Automated zoom helps maximize image detail while maintaining sensitivity. Researchers can focus on individual blots, gels, or regions of interest without manually repositioning samples or adjusting camera settings.
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 enable 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 imaging area enables researchers to capture up to four mini blots or gels in a single image, helping improve throughput, reduce imaging time, simplify side-by-side sample comparisons, and maintain a compact benchtop footprint.
Accelerate your work with fluorescent multiplexed western blots
Multiplex fluorescent western blotting enables simultaneous detection of multiple protein targets on a single membrane. This can help conserve samples, reduce experimental variability, and simplify comparison of target proteins and loading controls within the same experiment.
The iBright FL1500 Imaging System supports multiplex fluorescent western blot imaging with up to four fluorescent channels, enabling detection and analysis of multiple protein targets within a single experiment. Multiplex fluorescent detection can reduce sample consumption, improve experimental efficiency, and enable more accurate comparisons between targets and controls by capturing multiple signals on the same membrane.
The iBright FL1500 Imaging System includes multiple excitation and emission filter combinations to support common visible and near-infrared fluorescent dyes. Filter sets pre-installed in iBright FL1500 Imaging System 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 |
Advanced gel imaging capabilities
iBright imaging systems include accessories and imaging technologies designed to improve agarose gel imaging quality, reduce background fluorescence, and support safer nucleic acid imaging workflows.
Why use a green LED transilluminator?
iBright imaging systems use a green LED transilluminator to excite common DNA stains such as ethidium bromide and SYBR Green dyes without exposing samples to UV light. This helps support safer operation while preserving sample integrity for downstream applications.
Preserve sample integrity from 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.
Reduce mercury-containing bulbs and 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.
Improve imaging of E-Gel precast agarose gels
iBright Tray Adapters help optimize imaging of Invitrogen E-Gel precast agarose gels by improving gel positioning and reducing background fluorescence from the cassette label and frame. This helps produce clearer gel images and more consistent analysis results.
iBright Tray Adapters are designed to improve image quality when imaging E-Gel precast agarose gels by minimizing background fluorescence and helping properly position the gel within the imaging area.
Invitrogen iBright Tray Adapters for E-Gels. The Invitrogen iBright Tray Adapter for E-Gels, 11/22-well (panel A) and the Invitrogen iBright Tray Adapter for E-Gels, 48/96-well (panel B).
The iBright Tray Adapter for 48/96-well E-Gel precast agarose gels on the iBright turntable. The top component of the tray adapter (1) prevents fluorescence emitted by the E-Gel cassette (2) from reaching the camera detector. The bottom component of the tray adapter (3) centers the E-Gel cassette over the transilluminator (4) which is below the turntable (5) Light emitted by the green LED transilluminator shines through the open window in the bottom component.
Imaging with the iBright Tray Adapter for 48/96-well E-Gel precast agarose gels. Gel imaged without tray adapter (left) and with the tray adapter (right).
Related application note:
Support accurate quantitative western blot analysis
iBright imaging systems and iBright Analysis Software support total protein normalization workflows, enabling researchers to capture, quantify, and compare total protein signals and target protein expression using a single imaging and analysis platform.
Total protein normalization using the No-Stain Protein Labeling Reagent: Bolt 4–12% Bis-Tris Plus gels were loaded with HeLa lysate ranging from 10 to 50 µg. Proteins from the gels were transferred onto PVDF membranes using the Invitrogen iBlot 2 Gel Transfer Device with iBlot 2 Transfer Stacks, PVDF, mini. The PVDF membranes were labeled with No-Stain labeling solution, followed by immunoblotting for β-actin, GAPDH, and α-tubulin followed by goat anti-mouse Alexa Fluor Plus 680. (A) The blot was imaged using the iBright imager. (B) The iBright software was used to quantitate the total protein signal in the lanes. The linear regression value of the plotted data for the entire load range using the No-Stain Protein Labeling Reagent was determined (R2 = 0.9990), whereas the R2 values for β-actin, GAPDH, and α-tubulin were 0.8851, 0.9438, and 0.8332, respectively.
Related application note:
How-to videos
Watch short videos demonstrating key iBright imaging system capabilities, including automated image acquisition, image analysis, multiplex fluorescent imaging, and quantitative western blotting workflows.
iBright overview
Introduction to Invitrogen iBright imaging systems
Get an overview of iBright imaging systems and learn how they support western blot imaging, gel documentation, automated image analysis, and quantitation protein analysis workflows.
iBright how-to demonstrations
*Note that we make regular updates to instrument experience and function through firmware updates. The videos above accurately reflect the majority of instrument functions as featured, but some workflow and feature enhancements may not be reflected in these videos.
Frequently asked questions
Smart Exposure technology automatically evaluates the image signal and determines the optimal exposure settings for image acquisition. By selecting appropriate exposure conditions, it helps reduce manual optimization, minimize repeated exposures, and improve imaging consistency across experiments.
Smart Range HDR imaging combines multiple exposures to help capture both strong and weak protein signals within the same image, improving visualization and quantitative analysis across a wide dynamic range.
Yes. iBright imaging systems include integrated analysis tools that support densitometry, molecular weight estimation, total protein normalization, and protein quantitation workflows.
iBright imaging systems automate sample positioning, focus, zoom, and image alignment while using Smart Exposure technology to simplify image acquisition. This helps reduce manual setup, minimize repeated image captures, improve image consistency, and streamline western blot and gel imaging workflows.
iBright imaging systems support chemiluminescent, fluorescent, and colorimetric western blot imaging; protein and nucleic acid gel documentation; colony counting; In-Cell Western assays; and selected visible-light imaging applications such as TLC plates and plant tissues.
The iBright FL1500 Imaging System has 5 fluorescent imaging channels and can capture up to four channels simultaneously.
Yes. Available software tools support workflows that include electronic records management, audit trails, and user access controls.
iBright imaging systems support integrated image analysis using on-instrument tools and iBright Analysis Software. Available capabilities include densitometry analysis, molecular weight estimation, total protein normalization, colony counting, image annotation, and quantitative analysis for western blot, protein gel, and nucleic acid gel imaging. (Learn more about iBright Analysis Software).
For Research Use Only. Not for use in diagnostic procedures.