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For quantification of the migration of implanted OECs, longitudinal sections (10 µm) were cut in the horizontal plane.
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Placing cells labeled with different fluorescent markers in specific locations allowed quantification of the migration of cells between the layers.
In addition, a quantification plasmid allowed for quantification of the bacteria copy number.
Quantification of the migration rate in scratch assay showed significantly increased migration with TGF- β2 treatment, in parallel to spheroid assay results.
For quantification of migration, speed scram-shRNA TOM+ PNs (n=98 cells), 5-HT6R-shRNA1 TOM+ PNs (n=94 cells) and Cdk5/p35+5-HT6R-shRNA1 TOM+ PNs (n=95 cells) located in deep cortical layers were tracked using Metamorph.
Insert membranes were isolated following migration and stained with DAPI to allow for quantification of migrated cells (total magnification, 100×).
For quantification of invasion/migration, the total area of stained cells was measured using the Threshold command of MetaMorph 6.2.6 (MDS Analytical Technologies, Sunnyvale, CA, USA) and divided by the total surface of the membrane (% of filled area).
Quantification of the cell migration assay is shown in Fig. 3D.
(B and E) Quantification of the distance of cell migration in the direction toward the center of the wound.
Assays for quantification of cellular responses vary depending on the specific application and may include measurements of cell migration, overall cell integrity or secretion of specific ligands.
A standard curve for quantification of mycobacterial load was generated.
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