Figure 2
Figure 2. Autofluorescence analysis by deconvolution microscopy. (A) Small cell with few blue autofluorescent structures at the start of culture. (B) Same cell as panel A, by phase-contrast microscopy. (C) Intermediate size cell with many, paranuclear, blue autofluorescent structures after 7-day culture with TGF-β. (D) Same cell as panel C by phase-contrast microscopy: the cell is dendritic in shape. (E) Large cells with many autofluorescent structures that appear in part green, after 14-day culture with TGF-β. (F) Same cells as panel E by phase-contrast microscopy: the cells are markedly dendritic in shape. (G-H) Cell after 18-day culture with TGF-β; green autofluorescent structures are less numerous and less intensely fluorescent than after 14-day culture (compare with panel E). Bar represents 10 μm. (I-J) Spectra of autofluorescence emission after 1-day (black), 7-day (dark gray), 14-day (continuous light gray), and 18-day culture (discontinuous light gray). (J) The spectra were normalized assuming the respective highest peak as 100%. Live cells were mounted on slides with PBS; images were taken with ViSTA 1.9.3 software through a KAF261E detector (Kodak) inserted in a Hires IV camera (DTA) applied to an Eclipse TE-2000-E microscope with a 100×/1.3 NA oil objective (Nikon), and were processed for print with Photoshop 6.0 for Macintosh (Adobe).

Autofluorescence analysis by deconvolution microscopy. (A) Small cell with few blue autofluorescent structures at the start of culture. (B) Same cell as panel A, by phase-contrast microscopy. (C) Intermediate size cell with many, paranuclear, blue autofluorescent structures after 7-day culture with TGF-β. (D) Same cell as panel C by phase-contrast microscopy: the cell is dendritic in shape. (E) Large cells with many autofluorescent structures that appear in part green, after 14-day culture with TGF-β. (F) Same cells as panel E by phase-contrast microscopy: the cells are markedly dendritic in shape. (G-H) Cell after 18-day culture with TGF-β; green autofluorescent structures are less numerous and less intensely fluorescent than after 14-day culture (compare with panel E). Bar represents 10 μm. (I-J) Spectra of autofluorescence emission after 1-day (black), 7-day (dark gray), 14-day (continuous light gray), and 18-day culture (discontinuous light gray). (J) The spectra were normalized assuming the respective highest peak as 100%. Live cells were mounted on slides with PBS; images were taken with ViSTA 1.9.3 software through a KAF261E detector (Kodak) inserted in a Hires IV camera (DTA) applied to an Eclipse TE-2000-E microscope with a 100×/1.3 NA oil objective (Nikon), and were processed for print with Photoshop 6.0 for Macintosh (Adobe).

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