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Explore iBright Analysis Software capabilities and analysis tools for western blot and gel imaging.
Explore the western blot workflow
Analyze, quantify, and manage western blot and gel images with confidence
All iBright imaging systems include integrated on-board analysis software for routine image analysis directly on the instrument. The iBright Analysis Software extends these capabilities with desktop, cloud-based, and secure software options for advanced image analysis, collaboration, and data management. We regularly update our software to add features and to continuously improve the user experience. Together, these tools help researchers quantify protein expression, perform total protein normalization, generate publication-ready figures, and manage imaging data throughout the analysis process.
iBright software key capabilities include:
- Relative and absolute quantitation
- Total protein normalization
- Densitometry analysis
- Molecular weight estimation
- Colony counting
- Image annotation and reporting
- Image organization and data management
Available platforms for iBright Analysis Software
Our standalone iBright Analysis Software is a free software for organizing and analyzing blot and gel images captured on iBright imaging systems. Our iBright Analysis Software was designed to build upon the on-board analysis workflows, with more functionality and with the added convenience of multiple software formats depending on preference and need.
The standard iBright Analysis Software is available in desktop, cloud-based, and secure formats to support a variety of image analysis workflows.
- On-board—Analyze directly on the instrument
- Desktop—Perform flexible analysis offline
- Cloud—Access, review, and collaborate through Thermo Fisher Connect
- Secure—Supports workflows requiring 21 CFR Part 11 compliance
The table below summarizes the capabilities, supported platforms, and typical use cases for each iBright Analysis Software option.
Software option |
Typical use |
Platform |
On-board analysis |
Routine image analysis directly on the instrument |
iBright imaging system |
Desktop software |
Flexible image analysis independent of imaging instrument or internet connection |
Windows® and macOS® |
Cloud software |
Image access, analysis, and collaboration through Thermo Fisher Connect |
Web browser |
Secure software |
Image analysis and data management for workflows requiring 21 CFR Part 11 compliance |
Windows® and macOS® |
On-board analysis software
Perform routine image analysis directly on the iBright imaging system. On-board analysis software automatically identifies lanes and bands and enables densitometry, molecular weight estimation, quantitation, and normalization without transferring images to a separate computer. Furthermore, up to four blots or gels can be analyzed simultaneously, greatly increasing throughput.
Watch how on-board analysis is performed.
Analyze, organize, and document your data
iBright Analysis Software organizes image analysis into four main workspaces: gallery, adjust, analyze, and annotate. These dedicated workspaces help researchers manage images, optimize presentation quality, perform quantitative analysis, and prepare results for reporting.
Software workspace |
Function |
Features |
Gallery |
Explore and organize your images |
View, store, sort, search, select, import, export, and create reports |
Adjust |
Fine-tune and refine your images for meetings and presentations |
Rotate, straighten, flip, zoom, crop, invert, adjust contrast and gamma levels, apply false colors |
Analyze |
Gain a deeper understanding of the data |
Auto-analysis (automatically detect lanes and bands), manual analysis (user-defined), quantitation (relative or absolute), and normalization using total lane protein and housekeeping proteins |
Annotate |
Label your image |
Label analysis objects (Frames, lanes, bands & regions) through data table and/or add text, boxes, and arrows using annotation tools |
Manage your images with ease
The Gallery workspace can be sorted based on mode (application), date, file name, capture type, image information, and notes, so your data is readily retrievable and organized. Additionally, image preview is adjustable so you can scan through closely related data without opening each image individually.
Our iBright Analysis Software provides the capability to import proprietary image files from iBright imaging systems, iGlow Gel Documentation Systems (.iba), and Invitrogen E-Gel Power Snap Plus Electrophoresis systems (.g2i), as well as Invitrogen Power Snap Electrophoresis systems (.vit). Additionally, unmodified and uncompressed TIFF files from other imaging instruments can be imported into the iBright Analysis Software. This feature is beneficial for labs with multiple western blot and gel imaging instruments, including an iBright system, and wish to use a single program for all data analysis to help ensure consistency in the analysis process. The iBright Analysis Software offers users the flexibility to work with their existing data and leverage the comprehensive analysis tools and functionalities provided by our software.
Find, sort, and manage your images in the gallery workspace.
Analyze your data quickly
Image analysis can be simplified to only a few clicks in the Image Analysis workspace of the iBright Analysis Software. Automatically analyze up to four mini blots or mini gels at a time from a single image for increased throughput. Powerful algorithms are designed to consistently and accurately select and identify lanes and band boundaries for molecular weight estimation, densitometry, or normalization. Adjustments can be made to band display sensitivity to fine-tune the number of bands displayed for analysis. Images can also be analyzed manually if preferred.
Blots analyzed using automatic analysis feature. In this example, separate blots are identified with four independent analysis frames that can be edited and analyzed separately.
The line plot option in the manual analysis workflow allows users to obtain an intensity profile along a designated line or a drawn box. This tool can help researchers quickly assess the linearity of signal or confirm the desired outcome of an experiment. By generating a line plot, users can visually examine the distribution of intensities along the designated line, providing valuable insights into the experimental outcomes.
Lane plot analysis can help one quickly evaluate an experimental outcome via examination of the distribution of intensities along the designated line or drawn box.
Adjust Image. Image adjustment options are highlighted within the Image Analysis workspace, which can be used to straighten, crop, rotate, flip, or invert images according to need and preference.
Lane profile is the intensity profile plot of the selected lane. It is plotted with average pixel intensity of all pixels at that location or band if identified on the x-axis and pixel position on the y-axis. The chart also shows the non-specific background detected by the software in gray. By default, rolling-ball background subtraction is performed using an optimal rolling ball radius produced by the software for that image. This helps to more clearly view the band boundaries and more easily adjust the bands according to profile. Hovering over the profile graph provides values for band intensity and relative front.
Use lane profile to adjust bands and leverage rolling-ball background correction.
Label for a more complete picture
The Annotate workspace allows the user to label the image. All annotations will appear in magenta in the viewport. Annotations and lane labels added using the annotation table will be included in the exported image and all annotations are included in the image report.
Annotate workspace—lane and band labels. Lane labels were added through a data table, and remaining labels were added using annotation tools.
Bring it all together with a complete report
After analysis, view your images alongside your data report. Data can be filtered to view only desired parameters. Reports can be exported to PDF format, or the raw data can be exported to Excel software for further analysis.
Exportable report helps streamline post-imaging documentation.
iBright Analysis Software—Desktop application
The iBright Analysis Software desktop application is available for anyone to download and is compatible with both PC and Mac operating systems.
Make sure your desktop software is up to date! Download the latest version in addition to instrument firmware updates and patches.
iBright Analysis Software—Cloud-based
Images captured on the iBright imaging systems can be exported directly to Connect, our cloud-based platform. Conveniently access and analyze your images wherever an internet connection is available with our cloud-based software. Furthermore, with Instrument Connect in Thermo Fisher Connect, you can determine instrument status, firmware edition, and usage history, offering an extra degree of control over the monitoring of your investment.
Each Connect account offers 1 TB of cloud storage at no charge. Link user accounts on an iBright imaging system to a Connect account to directly upload data to a personal gallery on Connect.
iBright Analysis Software—Secure
iBright Analysis Software—Secure is a separate version of our iBright Analysis Software platform to support compliance with 21 CFR Part 11 FDA guidelines for Security, Auditing, and Electronic Signatures when connected to the SAE Administrator Console via the SAE server. The iBright Analysis Software—Secure desktop software can be utilized on computers running Windows or Mac operating systems, and can be configured to:
- Control access to data using account credentials
- Track user actions through audit logs
- Detect data tampering
- Sign electronic documents using electronic signatures
What is 21 CFR Part 11 Compliance?
The FDA released the Electronic Records and Signatures Rule, known as 21 CFR Part 11 in August 1997. This rule defines the requirements for use of electronic documents in place of paper documents. The law specifies the system elements, controls, and procedures that are necessary to help ensure the reliability of electronically stored records.
21 CFR part 11 compliance is composed of both procedural and technical requirements. Procedural requirements are the standard operating procedures instituted by the end user, and technical requirements are the functional characteristics of the compliance management software used. Satisfying the technical requirements does not guarantee 21 CFR part 11 compliance alone. Compliance is the consequence of the end user’s work process and systems used.
For technical details on how the Invitrogen iBright SAE Software Solution for 21 CFR Part 11 Support helps ensure workflows compliant to 21 CFR part 11, review the technical specifications document.
What is the iBright SAE Software Solution for 21 CFR Part 11 support?
To implement the iBright SAE Software Solution for 21 CFR Part 11 support on iBright imaging systems, you need the following components installed, activated, and communicating with each other:
- SAE Administrator Console Software: used with the iBright imager and Analysis Application Profile to configure the security, audit and e-signature settings for iBright imaging systems.
- iBright SAE License: used to activate the SAE settings for the iBright instrument and the iBright Analysis Software—Secure desktop software.
- iBright Instrument—SAE Mode: firmware mode that connects the instrument to the SAE Administrator Console.
- iBright Analysis Software—Secure: desktop analysis software that connects with the SAE Administrator Console.
iBright 21 CFR11 Software Solution* elements.
(1) Security, Audit and e-signature (SAE) Software (SAE Administrator Console, SAE Server, and SAE Application Profile (i.e. iBright imager and Analysis Software), (2) iBright instrument—SAE Mode (onboard software), (3) iBright Analysis Software—Secure (desktop analysis software).
*iBright SAE Software License is required for activation of the 21 CFR Part 11 support package.
SAE Administrator Console
The SAE Administrator Console is the tool used to configure security, audit, and e-signature settings to meet user-specific requirements for security, audit, and e-signature of a particular instrument and/or analysis software. The instrument and/or analysis software application profile (e.g. iBright imager and Analysis Software application profile) must be installed in the SAE Administrator Console before the SAE settings can be used on the instrument and/or analysis software. Once the application profile is installed in the SAE Administrator Console, an SAE server is used to connect to the instrument and/or analysis software to the SAE Administrator Console. By default, the SAE server is installed on the same computer as the SAE Administrator Console, which should be a network-connected computer configured with a static IP address.
Interface view from the SAE Administrator Console.
iBright SAE License
iBright SAE Software License for 21 CFR Part 11 is required to activate SAE Mode on the iBright imaging system and to activate the iBright Analysis Software—Secure desktop analysis software.
iBright imaging system SAE Mode
When SAE Mode is activated, the iBright instrument connects to the SAE Administrator Console via the SAE server. Security, Audit and e-signature settings, as defined in the SAE Administrator Console, are implemented on the iBright instrument. Features include restriction of unauthorized users to the system, password policies and expiration dates, defined user permissions and roles, tracking of all data changes through audit log, available audit reports, and on-board e-signature workflows. All iBright imaging systems can run in SAE Mode through purchase of the iBright SAE Software License.
Visuals from the iBright Imaging System interface showing how to access SAE mode.
How does the iBright SAE Software Solution for 21 CFR Part 11 Support work?
Functional characteristics of the Invitrogen iBright SAE Software Solution for 21 CFR Part 11 Support
Function |
Description |
System security |
Controls user access to an application by allowing creation of user accounts and defining user privileges through roles and password policies. |
Auditing |
Tracks actions performed by users, changes to the SAE module settings, and creation of audit reports. |
Electronic signature (e-signature) |
Determines if users are required to fulfill signature requirements before performing specific functions. |
Additional design features for each function are detailed below:
1. Access is restricted to authorized personnel via user ID and password.
Interface of the SAE Administrator Console showing new user account creation.
2. Passwords and user ID must expire after a user-defined number of days (typically 90 days).
3. Automatic logout after user-defined minutes of inactivity.
Interface of the SAE Administrator Console showing username and password policy settings.
4. ID and password must be re-entered to sign or modify record.
5. Must have a method to record and automatically report ‘forgery’ attempts (event notification).
6. Access and manipulation of various software functions are controlled through user role definitions.
7. Control access to specific areas and options in software with role settings.
Interface of the SAE Administrator Console showing user role settings.
Tracks actions performed by users and changes to the SAE settings. The SAE Administrator Console automatically tracks auditable actions silently (including, but not limited to, sign in/sign out, image acquisition, image exported, image deleted, file signed and report generation).
Interface of SAE Administrator Console showing the audit history tab.
(1) audit history available (action records, system configuration records, application object records), (2) application (SAE, iBright Instrument and Analysis Software, or other), (3) action, (4) date and user name (5) export in readable/printable format (PDF or .txt), (6) object ID.
With the auditing functionality, you can:
- Select to audit specific user actions and specify the audit mode (silent, optional or required)
- Generate reports for audited user actions and SAE module changes
- Generate reports on software or instrument actions and events
Committing auditable actions
Auditable actions can be committed onboard the iBright imaging systems instrument or in the iBright Analysis Software—Secure software. In order to track changes, changes must be committed and saved. The committed changes made can be viewed under the audit tab in the software application. Every commit creates a new version of metadata and is logged in the audit summary table in the audit tab. All committed actions are sent to SAE Administrator Console and logged under audit action records.
In the Commit workflow, select the Reason to commit from the selections displayed (or choose Other/Custom Reason). Enter comments as required and click on Commit.
iBright Analysis Software—Secure desktop software showing the reason to commit window and function. (1) Reason field (2) Option to add a custom reason (3) Comments.
iBright imager SAE Mode auditable changes screens, displaying reason to commit window and function.
(A) Reason field, touch “Blank reason” box to open audit reason screen (B) Select from options or “Other” to add a custom reason (C) Enter comments if needed and touch Save (D) Confirmation screen of changes committed.
Determines if users are required to fulfill signature requirements before performing specific functions.
Interface of the SAE Administrator Console e-signature settings.
(1) customize the meanings of e-signatures (2) customize the number of e-signatures required by meaning and role (3) type of data signed for a specific selected meaning.
With the electronic signature functionality, you can:
- Configure each e-signature event to require users with specific roles to sign
- Create separate meanings with customized signature requirements for each meaning
Signing files and generating signed reports
When all the changes are done and committed, the file can be signed as per signature rules set in the SAE Administrator Console. Signature requirements (number of signatures and role required) for the selected meaning will be listed as shown below. Once all signatures are obtained, the status of the signature rule will be updated in the e-signature table for the corresponding version found in the audit tab from unsigned to signed. A signature log will be added in e-signature summary table.
In e-signature workflow, select the applicable e-signature meaning. Required signatures and roles will be listed. Select the applicable role and enter credentials. When all signatures are obtained, the file will be fully signed and locked.
Note: iBright Analysis Software—Secure allows partially signed files to be shared across user computers in partially signed state. When all signatures are obtained, the file will lock. iBright Instrument e-signature workflows requires all signatures to be obtained before exiting the e-signature workflow.
Signature workflow in iBright Analysis Software—Secure. Select Signature Meaning from available drop down. In iBright Analysis Software—Secure, each required role is listed, and signatures can be provided by clicking on “unsigned” status link for designated role. Partially signed files can be sent to additional users using iBright Analysis Software—Secure to complete all signature requirements.
iBright Imager SAE Mode. In iBright Imager SAE Mode, signature requirements for the designated Meaning are listed under the user icons. All signatures must be obtained at the same time onboard the iBright.
A report can be created directly from the iBright imaging systems instrument or from the iBright Analysis Software—Secure desktop software as either unsigned or signed. The file must be fully signed by all required roles/signatures prior to generating a signed report.
Export of image analysis report from iBright Analysis Software—Secure desktop software.
Software how-to videos
How do I analyze western blot and gel images using Invitrogen iBright Analysis Software?
Learn how to use iBright Analysis Software to organize, edit, analyze, annotate, and export western blot and gel images. This introductory tutorial walks through the software interface and demonstrates the core workflows used for image analysis, molecular weight determination, band quantitation, and report generation.
You'll learn how to:
- Organize and manage images in the Gallery tab
- Optimize images using editing and adjustment tools
- Perform band quantitation and molecular weight analysis
- Annotate images and generate analysis reports
- Export images and analysis results
- 1:00—Brief overview of iBright imaging systems and iBright Analysis Software
- 4:40—Gallery tab overview (importing images, gallery organization, adding and removing images from tray)
- 7:23—Adjust tab overview (editing functions: crop, straighten, rotate, flip, invert, false color, contrast, show saturation)
- 19:15—Analyze tab overview (automatic analysis, analysis frames, adding and removing bands from analysis, resizing band frames, molecular weight analysis, data table, auto-enhance, lane profile)
- 36:32—Annotate tab overview (labeling images)
- 44:25—Generating and exporting an analysis report
- 47:42—Image export
- 51:30—Renaming image files
- 52:15—Question & answer session
How do I perform quantitative western blot analysis using Invitrogen iBright Analysis Software?
Learn how to perform quantitative western blot analysis using iBright Analysis Software. This tutorial demonstrates best practices for image quantitation, background correction, and normalization using housekeeping proteins and total protein normalization (TPN) to support accurate protein expression analysis.
You'll learn how to:
- Perform lane and band quantitation
- Apply background correction methods
- Normalize data using housekeeping proteins
- Perform total protein normalization (TPN) with No-Stain Protein Labeling Reagent
- Generate quantitative analysis reports
- 1:10—Overview of iBright imaging systems and considerations for achieving quantitative western blots
- 19:45—Brief overview of Gallery tab functionality and image selection for analysis
- 21:45—Brief overview of Adjust tab functionality
- 23:00—Analyze tab (auto-analysis, lane and band adjustments)
- 26:30—Normalization analysis with housekeeping proteins (HKPs)
- 31:30—Background correction (local background correction, lane profile, and rolling ball background correction)
- 37:50—Relative quantitation
- 39:30—Total lane protein (TPN) normalization with No-Stain Protein Labeling Reagent
- 43:40—Total lane protein (TPN) normalization “across the file” with membrane stain
- 50:10—Generating an image report
- 53:00—Question and answer session
Frequently asked questions
The iBright Analysis Software is a free image analysis application for western blot, protein gel, nucleic acid gel, and colony imaging workflows. It supports densitometry, molecular weight estimation, protein quantitation, normalization, annotation, and report generation to help researchers analyze and document imaging results.
Yes. The software supports automatic and manual densitometry analysis workflows for western blot and gel images.
Yes. Molecular weight estimation can be performed using molecular weight markers and integrated analysis tools within the software.
The iBright Analysis Software is available in multiple formats to support different workflows. All iBright imaging systems include integrated on-board analysis software. Desktop software is available for Windows® and macOS®, while a cloud-based version is available through Thermo Fisher Connect, providing browser-based access to images and analysis tools.
Yes. The iBright Analysis Software—Secure is designed to support workflows that require 21 CFR Part 11 compliance support.
The iBright Analysis Software supports densitometry analysis and relative or absolute quantitation workflows. Researchers can automatically identify lanes and bands, adjust analysis boundaries, and export quantitative data for reporting and downstream analysis.
Yes. The iBright Analysis Software supports total protein normalization and housekeeping protein normalization workflows to help improve the accuracy and reproducibility of quantitative western blot analysis.
Yes, The iBright Analysis Software can import 16-bit TIFF image files, generated by other imaging systems, helping users analyze images from multiple imaging workflows.
For Research Use Only. Not for use in diagnostic procedures.