ABSTRACTS – ORAL PRESENTATIONS - AMCA, spol. s r.o.

ounds on BD instruments only. Background staining levels (negative lymphocytes) were
also comparable. While we did see systematic platform related differences when using
hard dyed Rainbow beads, probably attributed to different filter sets used in collection
optics, they were improved by using single stained capture beads.
Conclusions: We show that polychromatic flow cytometry protocols can be standardized
even across different flow cytometry platforms developed by different makers. This opens
an opportunity to data exchange, inter-laboratory collaborations and computational data
analysis of multi centric studies regardless of flow cytometry equipment used. Variability
of the data introduced by instrument is lower than sample preparation variability.
Acknowledgements
Supported by UNCE 204012, GAČR P/302/12/G101, GAČR P/301/10/1877, NT/12425-4,
NT/14534
References
Kalina T. et al: EuroFlow standardization of flow cytometer instrument settings and
immunophenotyping protocols. Leukemia. 2012 Sep;26(9):1986-2010.
van Dongen J.M.M. et al: EuroFlow antibody panels for standardized n-dimensional flow
cytometric immunophenotyping of normal, reactive and malignant leukocytes.
Leukemia. 2012 Sep;26(9):1908-75.
P45. USE OF MICROFLUIDICS TO ACHIEVE NATIVE PHENOTYPE OF ENDOTHELIAL CELLS
Barbora Ambrůzová 1,2 , Hana Kolářová 1,3 , Martin Kubeš 1,2 , Lucia Binó 1,2 , Jan Víteček 1,3 ,
Lukáš Kubala 1,3
1
Institute of Biophysics ASCR v.v.i., Brno, Czech Republic; 258462@mail.muni.cz
2
Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech
Republic;
3
International Clinical Research Center – Centre of Biomolecular and Cellular
Engineering, St. Anne‘s University Hospital Brno, Czech Republic
Vasculature is a complex system composed of many cell types organized in a specific
manner to fulfill its physiological function. Vascular endothelial cells play role in many
physiological processes, e.g. maintenance of vascular tone and immune response (Choi,
2009; Giles, 2012).
The native phenotype of these cells is characterized by elongated shape, organization in
the direction of blood flow, tight contacts among them and remarkable glycocalyx layer
at the luminal surface. Such phenotype is however suppressed under standard static in
vitro cultivation (Henderson-Toth, 2012).
Our main focus is the inner layer of endothelial cells. In order to get native phenotype
of endothelial cells, HUVECs were cultivated under shear stress in a flow through system
operated by IBIDI pump. Flow conditions resulted in elongation of cells and longitudinal
orientation with flow. Further, elevated expression of glycocalyx-related genes was
detected using RT PCR after 3 and 7 days.
It can be suggested that microfluidic system based on IBIDI pump and µ-slides can
Analytical Cytometry VII 141