Images of pig kidney epithelial (LLC-Pk1) cells that were treated with both primary (anti-tubulin) and secondary (goat anti-rat FITC) antibodies for indirect immunofluorescence were taken at 10X magnification (Figure 1B & 1D) and 100X magnification (Figure 2B & 2D) under a fluorescent filter. Additional images were taken in phase contrast at 10X and 100X magnification (Figure 1A & 1C, Figure 2A & 2C, respectively). Additional LLC-Pk1 cells were treated in the absence of a primary antibody, (with buffer solution and secondary antibody) to create a control group for indirect immunofluorescence. These cells were imaged at 10X magnification (Figure 1C & 1D) and 100X magnification (Figure 2C & 2D) under a fluorescent filter. It can be visually discerned that there is a large difference in localization of fluorescent dye and ability to visualize tubulin structures between the two conditions. Figure 1B demonstrates a clear localization of fluorophores to the cells, specifically of tubulin structures around the nucleus. Figure 1D shows contrasting visual data, where fluorophores are seen indiscriminately binding to cellular structures as well as resting on parts of the slide that contain no cells. These differences are again highlighted, showing the direct localization of dye to microtubules, and random dying of miscellaneous cellular structures at a higher magnification in Figures 2B and 2D, respectively. Additionally, quantification of fluorescence was measured for both control and indirect immunofluorescence groups for all magnifications. It is apparent that the control group possesses a lower fluorescence intensity in localized tubulin areas than the group using the primary antibody for indirect immunofluorescence at 10X magnification (22.123, 46.028, respectively) and 100X magnification (31.249, 63.962, respectively) (Table 1).
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