iBright Imaging Systems Features - US (2024)

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 modeWhat 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

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

Fluorescent colonies (e.g. GFP expression)

Visible colonies (e.g. crystal violet stained)

Application Note: Colony Counting with iBright Imaging Systems

Qualitative visible imaging applications*

TLC-plates

Leaf sections

Images pictured forfluorescent western blots, stained nucleic acid gels, colorimetric stained protein gels, GFPexpressing 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 forE-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.

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).

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 fromlane-to-lane to better understand the results.

Explore more: Application Note - Optimizing Invitrogen™ E-Gel™ precast gel imaging and analysis using the Invitrogen™ iBright™ Tray Adapters for E-Gels and deconvolution software

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.

Interface features

  1. Sign in: create a new user account of sign in to an existing account
  2. Gallery: access previously captured images
  3. Mode selection: dropdown menu to select imaging modes (chemiluminescent blot, fluorescent blot, nucleic acid gel, protein gel, and universal mode)
  4. Camera lock: maintain camera settings across multiple samples, locks exposure time, zoom, and transillumination settings while disabling mechanical auto-rotation
  5. Help: on-board help and information
  6. Settings: access general instrument settings, configurations, and service tools
  7. Drawer: open or close the sample drawer
  8. Exposure dial: select or turn the dial to set manual exposure time or fine-tune exposure time
  9. Capture: acquire images using exposure time set by user or instrument
  10. Edit channels: open multiple channel options to review and edit each channel
  11. More options: open image adjustment options
  12. Live view: go back to live-streaming sample view
  13. Analyze: initiate analysis workflow using current images
  14. Export: edit image information and export current images to destinations of choice
  15. Trash: delete current images
  16. Split screen: select one-window or two-window image view
  17. Color option: select color or grayscale image view
  18. Image view window: display selected image in the window for user view and interaction

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.

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.

Application Note: Expand dynamic range with iBright Imaging Systems’ Smart Range™ High Dynamic Range (HDR) technology

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 to 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.

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,iBrightImaging Systems andiBrightAnalysis Software provide multiple quantitation and normalization options to monitor or mathematically compensate for experimental or sample variability.

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iBright Imaging Systems Features - US (2024)

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