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. 2023 Jul 28;24(15):12139. doi: 10.3390/ijms241512139

Figure 5.

Figure 5

Figure 5

(a–i) Representative maximum intensity projection confocal microscopy images of immunocytochemistry staining of ADSCs in 3D with respective positive and negative staining controls for each antibody marker CNPase, NF200 and GFAP. Cells were imaged with a Nikon A1R inverted microscope using a S Plan Fluor LWD 20× 0.7NA objective. Fluorescence was captured with a laser at 405 nm excitation and PMT detector (425–475 nm) for DAPI (blue), and 488 nm excitation and GaAsP detector (500–550 nm) for AlexaFluor488-conjugated secondary antibodies (green). Scale bar = 100 μm. Please note that control cells are smaller than ADSCs; (j–l) immunocytochemistry marker expression for 3D cells. Marker expression was measured from sum intensity projections of wide-field fluorescence images of the whole 3D plug and is displayed as the fraction of the percentage area of FITC (AlexaFluor488-conugated secondary antibody) over the percentage area of DAPI-labelled nuclei. No statistical significance p > 0.05; statistical significance *, p ≤ 0.05; statistical significance **, p ≤ 0.01; statistical significance ***, p ≤ 0.001; statistical significance, ns: not significant.