M3 Vision
A software for the analysis of Biospace Lab images

Combine efficiency and precision for quantification, kinetics analysis and export

M3 Vision Features

The following examples illustrate different functionalities of the software:

Kinetic analysis

Three simple steps bring you to the temporal information of the signal:
  1. Create the region(s) of interest for which you want the kinetics. You can get the kinetics of the entire image, or of different ROIs. You can choose if you want each ROI to be considered separately (one curve per ROI) or if you want the kinetics of the union of the ROI.
  2. Select the time resolution.
  3. Choose the unit for quantification.
You are now just a click away of the kinetics!


Autofluorescence unmixing

The model

The autofluorescence unmixing process provided by M3 Vision relies on a simple, yet often efficient model, with the two following hypotheses:
  • Illuminating with the fluorophore's excitation wavelength gives signal mixed with autofluorescence
  • Illuminating with a lower wavelength ("background") gives autofluorescence of relative same intensity distribution as the previous autofluorescence
To unmix the autofluorescence and isolate the signal, we only need the ratio of intensities of both autofluorescence signals. This requires to compare on the intensities of autofluorescence on the two images. After normalization by this factor, the background autofluorescence is substracted from the acquisition.


The process


The process is very easy and relies on the three following steps:
  1. Creation of one or several dots, areas or lines on the image where there is only autofluorescence
  2. Selection of those objects
  3. Click on the "Build AF Substracted" button to get your unmixed images
Here's your signal!




Excitation image: the signal is mixed with autofluorescence


Background image: the back of the animal is selected, as we know a priori that the signal lies in the head of the rat. Without this knowledge, the selection of the autofluorescent area could have been done on a control animal.


Rat with Cy5 fluorescence in the brain.

Unmixing of the autofluorescence results in a clear isolation of the signal.

Sequence creation

The real time capacity of the system opens the way to dynamic recordings of the signal. The analysis software makes it thus possible to create dynamic sequences in order to observe the changes of the signal localization, intensity or distribution with time.
The creation of such a sequence requires you to define the following parameters:
  • Duration of each frame
  • Start and end point of the part of the acquisition to convert to a sequence
  • Signal and photography display parameters (min, max, smoothing)
Creating a sequence helps you visualize and quantify the evolution of the signal in the animal. You can export the sequence into a avi video file.


Automated creation of ROI

Detection and localization of signal sources, such as metastases for example, is one of the key applications of the Photon Imager. To ease the analyze and interpretation of the image, M3 Vision includes, among its numerous features, the possibility to identify, with the creation of regions of interest, all the signal sources in the image over a simple threshold. You can thus easily detect all tumors - primary tumors as well as metastases - that have a signal over a given threshold, and compare different animals, or animals at different time points with the same parameter.

The tool, called "magic wand whole image", is very simple. It proposes, by default, a threshold corresponding to the "min" parameter you have selected for display, to select ROI on the visible signal sources. You can adapt it to correspond to the threshold you want. To visualize the regions of interest corresponding to the current threshold, click on "Apply". If you are not satisfied with the created ROIs, you may change the threshold and click again on "Apply": the ROIs automatically adapt. When you obtained satisfactory ROIs, click on "Exit".

The creation of ROIs helps you to quantify the activity of the sources (quantitation report) , analyze their kinetics (time profile) , and export the quantification and the kinetics to Excel .



Export!

Images export

M3 Vision offers different export formats for your images. Export your images to the tiff format for further quantification with other softwares. For illustration, you have the choice between jpg, bmp and an illustration tiff format.

Quantification, kinetics, spatial profiles

  Export in a single click all your data to Excel or to HTML. Curves will be plotted automatically in Excel. Export of the quantitation report will also include a snapshot of the image, with the LUT, and all the properties of the acquisition.

And how about my dynamic acquisitions?

Sequences and in actio acquisitions can of course be recorded in avi video files. You have access to many parameters (min and max of the LUT, intensity transform functions, smoothing, photographic image brightness and contrast, etc.) to obtain the optimal quality for your videos.