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The difference in depth arising from cell vs. chamber analysis was found to be small.
To test this idea, we used a chemotaxis assay with Jurkat T cells using a modified Boyden chamber analysis.
In vitro Boyden chamber analysis of NSC tropism to tumor chemo-attractants in conditioned media collected from serum-starved adherent tumor cells at 24 h and 48 h showed no significant difference in migratory properties between FE-Pro labeled and non-labeled NSCs (P>0.05) (Fig. 3A).
At d 160, segments of ileal mucosa were rinsed with cold sterile saline then either placed into Ringer buffer for Ussing chamber analysis or into RNA later (cross-section of tissue) at 4°C for 24 h and then maintained at −20°C for later RT-PCR analysis.
Adjacent segments of ileum were rinsed with cold saline and either placed into Ringer buffer (composition in mmol.L−1: Na+ 145, Cl− 128, PO43− 0.32, Ca2+ 2, Mg2+ 1, HCO3− 25, SO42− 1, K+ 6.3) at 4°C for immediate Ussing chamber analysis, or into RNA later (Applied Biosystems, Courbaboeuf, France) (cross-section of tissue) at 4°C for 24 h and then maintained at −20°C for later RT-PCR analysis.
Results from Ussing chamber analysis are reported in Figure 1.
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In the first study 20 piglets were divided in 2 groups and received either a basal diet or the basal diet supplemented with OA + PB (5 g/kg) for 2 weeks post-weaning at the end of which ileum and jejunum samples were collected for Ussing chambers analysis of trans-epithelial electrical resistance (TER), intermittent short-circuit current (ISC), and dextran flux.
Self-supported oxide disks of approximately 0.5 cm diameter were loaded into the chamber for analysis.
The total pressure in the main vacuum chamber during analysis was typically 4 × 10−7 Torr.
After quenching, the sample was taken out from the MBE chamber for analysis using a Shimadzu SPM 9500 atomic force microscopy (AFM) (Shimadzu Scientific Instruments, Japan) in tapping mode.
A mass spectrometer linked directly to the sample chamber facilitated analysis of gaseous products evolved from the materials as a consequence of irradiation.
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