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Determine the Immuno-phenotype of AdMSCs by Countstar FL

Immuno-phenotyping analysis is a typical experiment performed in cell related research fields to diagnose various diseases (autoimmune disease, immunodeficiency disease, tumor diagnosis, hemostasis, allergic diseases, and many more) and disease pathology. It is also be used to test the cell quality in various cell diseases research. Flow cytometry and fluorescence microscope are the routine analysis methods in cell diseases research institutes used for the immuno-phenotyping. But these analysis methods can either provide images or data series, only, which may not meet the strict approval requirements of the regulatory authorities.


M Dominici el, Cytotherapy (2006) Vol. 8, No. 4, 315-317



Identification of Immuno-phenotype of AdMSCs

The immunophenotype of AdMSCs were determined by Countstar FL, AdMSCs were incubated with different antibody respectively (CD29, CD34, CD45, CD56, CD73, CD105, and HLADR). A signal-color application procedure was created by setting Green channel to image PE fluorescence, plus a bright field. Bright field picture reference segmentation was applied as a mask to sample the PE fluorescence signal. The results of CD105 were shown (Figure 1).


Figure 1 Identification of Immuno-phenotype of AdMSCs. A. Bright Field and Fluorescence Image of AdMSCs; B. CD Marker Detection of AdMSCs by Countstar FL



Quality control of MSCs – validating results for each single cell



Figure 2 A: The Countstar  FL results were displayed in FCS express 5plus, gating the positive percentage of CD105, and overview table single cells. B: Adjusted gating to the right side, the images of the single cell table shows those cells with high expression of CD105. C: Adjusted gating to the left side, the images of single cells table shows those cells with low expression of CD105.



Phenotypical Changes during Transport


Figure 3. A: Quantitative analysis of the positive percentage of CD105 in different samples by FCS express 5 plus software. B: High quality images supply additional morphological information. C: Validated results by thumbnails of each single cell, the FCS software tools divided the cells into different




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