The Reference Cell™, the new paradigm
in cell quantification
By introducing cell normalization and repeatability, two elements that were formerly lacking in cell quantification, CYTOO adhesive micropatterns make the Reference Cell
a dream come true...
Without changing your assay.
View the complete protocol to build a Reference Cell here:
Dots represent center positions of nuclei (in green) and centrosomes (in red) of 65 cells.The CYTOO approach to cell quantification
• Create a Reference Cell
As in biological tissue, cells cultivated in a defined microenvironment on a given CYTOO micropattern adopt the same shape and architecture, down to the position and morphology of their organelles and cytoskeleton.
Mapping a Reference Cell becomes possible.
• Gain tremendously in meaning and sensitivity
The signal to noise ratio significantly increases when summing the signal over just a few dozen cells.
• Achieve unlimited co-localization assays
You can use as many different labels as you need whatever their wavelengths, and obtain a composite image of them all.
4 cells on crossbow micropatterns.
Reference Cell obtainedfrom 74 stacked cell images (cortactin labeling).
The CYTOO process: cell quantification made easy
1 Step one: prepare
Prepare a CYTOO chip for each label, remember you can use as many labels as you need. Let cells sediment and adhere to the CYTOO micropattern arrays. Fix and label cells.
2 Step two: acquire
Scan the surface of each array with the indicated pitch between micropatterns.
Over each micropattern, acquire a high resolution image of each cell at the labeling wavelength. Create a stack of images and discard from the stack images of micropatterns with 2 or more cells or images where the cell has not spread out completely on the micropattern.
3 Step three: calculate
For each stack of images, calculate the average intensity of each pixel over the stack to obtain the average spatial distribution of the label of interest.
A color-coded rainbow intensity range can be used to highlight the main sites of the distribution.
Overlay the averaged images to obtain the Reference Cell.
Immediately observe colocalization.
4 Step four: compare
Carry out all your test points in the same manner and compare them to the Reference Cell.
100 µm or 130 µm between each micropattern.
