Pre-clinical research

Quantify your data in real ti me

Cellvizio^®Lab proprietary software, ImageCell™ provides patented, on-the-fly image reconstruction of the high-definition image data supplied by the LSU.

ImageCell™ 3.8

ImageCell™ enables you to go beyond imaging and to quantify your results in many areas.

Below are some examples of the quantification features offered by ImageCell™:

Vessel Detection™

Advanced Mosaïcing™

 Kinetics Analysis

Permeability Measurement

Vessel Detection™ Tool (optional)

With the Vessel Detection dedicated module, Cellvizio users will be able to easily compute a sophisticated segmentation of the micro-vessels, and extract a number of key measurements such as the Functional Capillary Density (FCD), the distribution of diameters, or the total cumulated length of the vessels.

The data can further be exported to be used in third party applications when needed. The following sections describe the different steps of the process: how to perform the segmentation, what quantitative analysis to get from the segmentation, and fnally how to export the data.

As for the rest of the ImageCell™ software, this module was designed to be as user-friendly as possible, with a minimal number of parameters to be set by the user, and offering the highest level of information.
 

Advanced Mosaïcing™ Tool (optional)

A mosaic is an optimized arrangement of successive frames within a sequence, which provides an enlarged view of the tissue.
Two tools are available:
• the Live Mosaicing: an online stitching tool
• the Post-processing Mosaicing: an ofine complete mosaicing tool with tissue distortion correction

The Live Mosaicing allows to get a real-time display of the draft mosaic built from the previous frames acquired together with the acquired image. Up to 200 frames can be stitched together, using an advanced  real time algorithm. Those frames are stored in a buffer. The acquisition frame rate will remain the same as in Image Only display mode. The right side of the screen displays the current image.

The left side of the screen displays the live mosaic built from the previous frames. The image enclosed in the red box is the current frame, displayed also on the right side. The Live Mosaicing mode is a great tool to visualize the stability of the image acquisition.

When the algorithm is unsure about the continuity of the mosaic or the stability of the image, it starts again by fushing the bufer.  Using the Live Mosaicing mode will ensure better high resolution mosaic with the post processing mosaicing tool.

Kinetic Analysis

The Kinetic Analysis Tool helps you study the kinetics of your signal intensities simply and fast, with only three steps:

1) Draw the regions of interest (ROI) on the sequence
2) Assign one or several functions of signal intensity to each ROI (Mean, Max, Min, Median)
3) View and handle the graph displaying the chosen statistics along time. It is then possible to export both the graph and the statistics.

The plots below (right) show signal kinetics of the ROI that appear on the Cellvizio^® sequences (left). They illustrate two applications of calcium imaging, respectively:

- Neuronal activation following visual stimulation in the cat cortex

- Spontaneous activation of neurons in the chick embryos

Vasodynamics of tumoral vessels

Constriction and dilation of tumoral vessels
upon carbogen and air breathing

Opposite: Constriction and dilation of tumoral vessels. The vasodynamics of a tumoral vessel was observed upon inhalation of carbogen by the mouse. On the first two images, the vessel is dilated while the mouse is breathing carbogen: we observe an increase of its diameter. After removal of carbogen, the vessel constricts back to its original size. The diagram (bottom of the figure) shows the vessel response to the sequence carbogen - air - carbogen.

Courtesy of Anne-Carole Duconseille, Charles A. Cuenod and Olivier Clément, Descartes Image, Small Animal Imaging Facility, Université Paris V, Paris, France

Permeability Measurement

Effect of histamine on vessel permeability.
Field of view: 400 x 280 µm

Blood extravasation is very easy to record and to quantify with Cellvizio^®.

Using the procedure that consists in dying the blood plasma, the fluorescence intensity of a vascular zone will be directly related to extravasation.

Thus, a study of the histogram or simply of the average intensity of a region of intesrest (ROI) can reveal the evolution of the quantity of plasma leaking towards the region of interest. Thanks to the features of ImageCell™, this analysis is immediate.

This diagram shows an example of this type of study. The extravasation generated by histamine is measured on the muscular tissues of a mouse. On the first image, a region of interest excluding the vessels and the capillaries is defined. The corresponding histogram therefore shows a signal with a very low level.

The second and third images show a significant raise of the fluorescence intensity of the ROI, the curve has moved to the right. Since the protocols requires blood plasma to be dyed with fluophore, the event observed allows the quantification of the extravasation on the analysed zone.
BSA-FITC can be an appropriate fluophore for this protocol.

For more information, please consult the following publications suivantes: E. Laemmel, M. Genet, G. Le Goualher, A. Perchant, J.F. Le Gargasson, E. Vicaut "Fibered confocal fluoresence microscopy (Cell~vizio) facilitates extended imaging in the field of microcirculation." (2004)
Journal of Vascular Research 41 (5): 400-411 

To be found here