Normalize longitudinal IVIS images
Apply one shared radiance minimum, maximum, log or linear scale, colormap, and color bar across quantitative panels.
Free browser-based BLI workflow
IVIS bioluminescence analysis and longitudinal figure building
SimpleBLI turns supported Living Image acquisition folders into consistently scaled IVIS panels, ROI measurements, time-course plots, CSV data, and publication-ready figures—without installing desktop software.
What SimpleBLI is built to do
SimpleBLI is a focused companion for the part of a BLI study that often becomes repetitive: arranging animals across time points, keeping the display scale consistent, calculating comparable ROI summaries, and exporting one clean figure.
Measure BLI regions of interest
Keep measurement ROIs separate from display annotations and export total-flux estimates, mean radiance, maximum radiance, and CSV data.
Plot signal over time
Compare animals, controls, and treatment groups using dated time points, group-colored traces, and mean or median summaries.
Build publication-ready BLI figures
Arrange individual animals or whole scans, add labels and a shared color bar, and export the displayed figure as a 300 dpi PNG.
How to analyze longitudinal IVIS bioluminescence data
This workflow keeps quantitative measurements and visual figure assembly connected while making the distinction between calibrated and rendered-only data explicit.
- Import acquisition foldersDrag in a study folder or select acquisitions containing Living Image metadata and luminescent TIFF data.
- Confirm quantitative statusSimpleBLI labels acquisitions as quantitative only when it can recover a usable calibration coefficient and exposure metadata.
- Define animals and groupsCrop animals, name subjects, assign control or treatment groups, and organize scans into dated time points.
- Normalize all panelsUse one robust or manually specified radiance range so color differences mean the same thing throughout the figure.
- Measure and plot ROIsChoose total-flux estimate, mean radiance, or maximum radiance and inspect individual or grouped time-course traces.
- Export figures and dataDownload the figure or plot as a 300 dpi PNG and export ROI measurements for downstream statistics.
Living Image and IVIS file compatibility
Use the original acquisition folder whenever quantitative comparison matters. A screenshot or pseudocolor export may look correct but does not necessarily contain the calibration needed to recover radiance.
| Input | How SimpleBLI uses it | Quantitative? |
|---|---|---|
ClickInfo.txt + luminescent.TIF | Reads acquisition metadata, raw CCD counts, exposure, binning, read bias, field of view, f-number, and the matching calibration coefficient. | Yes, when the required metadata and coefficient are available. |
photograph.TIF | Uses the white-light photograph as the anatomical background under the luminescent signal. | Supports the overlay; radiance comes from the luminescent data. |
| Standalone TIFF, PNG, or JPEG | Imports the file as a rendered panel for layout and labeling. | No. Rendered-only images cannot be rescaled quantitatively. |
.amix or .aurax | Native Living Image analysis and Aura project files are ignored rather than interpreted incorrectly. | No. Export supported source data or images first. |
Compatibility note: IVIS and Living Image exports vary across instrument generations and software versions. Confirm a representative acquisition in SimpleBLI before moving an entire study, and retain your original data and vendor-software results.
SimpleBLI vs Living Image vs Aura
These tools overlap, but they are not interchangeable. The best choice depends on whether you need instrument acquisition, advanced vendor analysis, legacy import, or a quick browser workflow for consistent longitudinal figures.
| Capability | SimpleBLI | Living Image | Aura |
|---|---|---|---|
| Primary role | Browser-based 2D longitudinal analysis and figure assembly. | IVIS acquisition and vendor analysis, including broader 2D, 3D, and applicable CT workflows. | Spectral Instruments acquisition and analysis for BLI, FLI, and X-ray. |
| Installation | No install; opens in a modern browser. | Vendor desktop software. | Free downloadable analysis software for Mac and PC. |
| Living Image data | Reads supported raw 2D acquisition folders with calibration metadata. | Native format and workflow. | Vendor states that Aura can import legacy Living Image files. |
| Shared-scale longitudinal figures | Core workflow: normalize panels, arrange animals/time points, plot groups, export figures. | Provides longitudinal analysis and publication-figure capabilities. | Provides visualization and analysis; workflow depends on imported data and Aura features. |
| 3D, CT, or spectral unmixing | Not supported. | Available in applicable IVIS workflows. | Use Aura’s documented vendor capabilities for supported instruments and data. |
| Best fit | Researchers who already have 2D scans and want a simple, repeatable path to figures and plots. | Labs needing native IVIS acquisition and full vendor analysis. | Labs using Spectral Instruments systems or needing Aura’s free desktop analysis and legacy import. |
Bottom line: SimpleBLI is an alternative workflow for a specific job, not a replacement for every Living Image or Aura feature. Keep vendor software for acquisition and specialized analyses; use SimpleBLI when the goal is fast, consistent 2D longitudinal presentation and measurement.
Total flux vs average radiance in IVIS analysis
These measurements answer different questions. Record the metric, units, ROI method, and any background or normalization procedure in your analysis notes and figure legend.
Radiance
photons/s/cm²/srCalibrated signal per unit area and solid angle. Radiance is used to map BLI intensity and set a common display scale across quantitative panels.
Average radiance
photons/s/cm²/srThe mean radiance of pixels inside an ROI. It describes typical surface brightness but can change when the ROI boundary changes.
Total flux
photons/sSignal integrated over the ROI. Living Image documents total flux using radiance, ROI area, and a solid-angle term, making it useful for whole-signal comparisons.
SimpleBLI total-flux caveat: SimpleBLI reports a reproducible estimate calculated from radiance and pixel area with the steradian term treated as 1 sr. Relative comparisons made with identical acquisition settings are the intended use. Absolute values can differ from Living Image because of its instrument-specific solid-angle convention; validate the metric before using it as a direct substitute.
IVIS bioluminescence analysis FAQ
Concise answers to common questions about free BLI software, Living Image files, Aura compatibility, ROI measurements, normalization, privacy, and publication figures.
Is SimpleBLI free?
Yes. The analysis and export tools are free, and there is no installer or license key required. You only need a paid plan if you want to save larger amounts of experiment data in private cloud storage.
Can I analyze Living Image data without installing Living Image?
For supported 2D acquisition folders, yes. SimpleBLI reads ClickInfo.txt and luminescent.TIF directly. Keep Living Image for native project editing, acquisition, 3D, CT, spectral unmixing, and other specialized workflows.
What files are required for quantitative IVIS analysis?
A supported acquisition needs ClickInfo.txt plus luminescent.TIF. photograph.TIF is optional but provides the white-light background. Quantitative status also depends on usable calibration metadata.
Does SimpleBLI open Aura or AuraX files?
No. Native .aurax files are not opened. Aura itself is free downloadable analysis software and can import legacy Living Image files according to Spectral Instruments. SimpleBLI can use supported raw Living Image folders or rendered image exports.
Is SimpleBLI a Living Image alternative?
It is an alternative for a focused subset of work: supported 2D data import, longitudinal normalization, ROI summaries, plots, and figure assembly. It is not a complete replacement for Living Image acquisition or advanced analysis.
Can SimpleBLI normalize every IVIS image to the same scale?
Yes. Quantitative panels can share one minimum, maximum, logarithmic or linear mapping, colormap, and color bar. That prevents each image from using a visually misleading independent range.
Can I perform IVIS ROI analysis?
Yes. Measurement ROIs report pixel count, area, mean, minimum, and maximum radiance, a documented total-flux estimate, and signal above an optional threshold. Measurements export as CSV.
Can I use ImageJ for IVIS images?
ImageJ is useful for general image processing, but a rendered PNG or screenshot does not automatically preserve IVIS radiance calibration. Use original calibrated source data for quantitative work and validate any cross-software workflow.
Does SimpleBLI upload my scans?
Signed-out experiments stay in browser storage on the current device. Signing in adds private account-backed cloud backup. Review your institution’s data-handling requirements before using any web application.
Can I make publication-ready IVIS figures?
Yes. SimpleBLI exports the displayed figure and plot as 300 dpi PNG files, with shared scales, color bars, labels, treatment colors, and individual-animal or whole-scan layouts.
Does it support fluorescence, 3D DLIT/FLIT, or CT?
No. SimpleBLI is currently focused on 2D bioluminescence workflows. Use the relevant vendor software for acquisition, fluorescence calibration, tomography, spectral unmixing, multimodal registration, or CT analysis.
Which metric should I plot over time?
Use a metric that matches the biological question and keep the ROI method consistent. Total flux estimates whole-ROI signal; average radiance describes mean surface brightness; maximum radiance emphasizes the brightest pixel region.
Sources and terminology
This guide distinguishes SimpleBLI’s tested behavior from vendor capabilities and uses established IVIS measurement terminology.
- Revvity Living Image software overview — vendor description of IVIS analysis and longitudinal workflows.
- Revvity technical note: Drawing ROIs for 2D Bioluminescence Imaging — measurement ROI guidance and units.
- Spectral Instruments Imaging: Aura software — vendor information on free analysis software and Living Image legacy import.
IVIS and Living Image are trademarks of their respective owners. Aura is a product of Spectral Instruments Imaging. SimpleBLI is an independent Simple Lab Tools application and is not affiliated with or endorsed by those vendors.
Build your first longitudinal BLI figure
Open SimpleBLI, create a free experiment, and drag in a supported Living Image acquisition folder. Your original files are not modified.
Open SimpleBLI free