Immuno-Fluorescence SEM
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Figure 9: Fluorescence materials used as standards. (A, D) Fluoresbrite carboxylate microspheres (Polysciences, Inc., Warrington, PA), ex. 529 nm. (B, E) Y2O3:Eu red phosphors (Oxnica Inc., Mountain View, CA). (C, F) GFP isolated from Aequorea forskålea [28]. (A, B, C) Electron images. (D, E, F) CL images.
the morphological/protein health of a cell and relevance of this to the failure of certain intercellular processes. We might also find it being applied to the tracking of cell growth and cell placement in tissue reconstruction with the use of cloned fluorophores lighting up new daughter cells. We look forward to the time when all the known surface proteins of the monocyte, as well as other cells, can be mapped at various stages of cellular differentiation, action, and life.
Conclusions We have shown the value of cathodoluminescence in
tracking the transfer of material from one cell to another, the placement of exogenous material on a cell, and the rearrangement of molecular concentrations on a cell surface. Exosomes, isolated from a U373 glioblastoma tumor culture and incubated with human monocytes, were shown to bind to the veils of the monocytes, in some cases retaining individual exosomal structure, and, in other cases, fusing into extended areas of the cell. In addition, differences in the overall tumor cell coverage by the surface protein EGFr and the concentration of this protein on developing exosomal structures were captured by cathodoluminescence. We were also able to show that the monocyte protein CD14 is found predominantly on the edges of the cell membrane veils. Future studies with other proteins relevant to the function of these cells will give us a strong footing for exploring the role of monocytes in tumor control.
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