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Immunofluorescent Staining of Cell lines with Conjugates and Quantification of Signal.

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posted on 2016-06-23, 07:05 authored by Steve Knutson, Erum Raja, Ryan Bomgarden, Marie Nlend, Aoshuang Chen, Ramaswamy Kalyanasundaram, Surbhi Desai

(A) Immunofluorescent images of cell lines stained with various conjugate samples. Two putative CEA-positive cell lines, BxPC-3 and MCF-7, and two control CEA-negative cell lines, HeLa and HepG2, were seeded at 1x104 cells/well in 96-well collagen-coated plates. Cells were immunofluorescently stained with approximately 2ug/mL of α-CEA-680 or α-CEA-680-PTX where noted (red). Nuclei were counterstained with Hoechst dye (blue). Images were obtained on an ArrayScan VTI under 20X magnification and arranged with the HCS View software, Pxlr editor, and Microsoft PowerPoint. (B) Quantification of Immunofluorescent Staining. BxPC-3, MCF-7, HeLa, and HepG2 cell lines were seeded at 1x104 cells/well in 96-well collagen-coated plates. Cell lines were probed with α-CEA-680 (red lines), α-CEA-680-PTX (blue lines), IgG-680 (green lines), or IgG-680-PTX (purple lines) at concentrations of 0 – 60ug/mL. Fluorescent signal intensity was quantified with the ArrayScan VTI and vHCS Scan software. Data points represent average fluorescent intensity (n = 3000 cells), and error bars denote standard error. Error bars are present at every data point, but are very small in some cases.

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