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In addition, a further sample was also collected of cells passed through the laser under pressure, but not sorted, to act as a control for the effect of the pressure on the cells.
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In control samples collecting of cells was started immediately after addition of SWCNTs.
Cells number was directly measured on the whole titanium disk to avoid any loss of cells during trypsinizing and collecting of the cells [ 26].
Five images each were collected of non-treated ES cells, control RNAi-treated ES cells and CTCF RNAi-treated ES cells.
To minimize the number of activated T cells that express high levels of CD25, we only collected 1% of cells with the highest CD25 expression.
Inside of the ciliary tuft a bunch of long cilia (Figure 5C) were found to be connected to collecting ducts of cells of the protonephridial system as seen in the magnification showed in Figure 6A.
After 3 days the medium was collected, cleared of cell debris by centrifugation, concentrated approximately 10-fold through a Centriplus YM-10 column (Millipore, Bedford, MA).
The number of apoptotic cells was estimated after incubation of an aliquot of collected living cells with PE-labelled Annexin-V (BD Pharmingen) and with 7-AAD.
Purity was assessed by flow cytometry with >90% of collected cells routinely bearing an NK cell phenotype of NKG2A+CD8+CD3-.
However, due to the low number of endothelial cells collected (mean of ∼97 cells in the current study) and the subsequent low RNA yield these types of material have never been practically accessible for global gene expression profiling.
Open image in new window Open image in new window Figure 2 Single cell transcriptome analyses reveal neuronal property of collected target cells.
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