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. 2010 Oct 18;11(15):2092-5.
doi: 10.1002/cbic.201000419.

Live-cell imaging of cellular proteins by a strain-promoted azide-alkyne cycloaddition

Kimberly E Beatty  1 , John D Fisk, Brian P Smart, Ying Ying Lu, Janek Szychowski, Matthew J Hangauer, Jeremy M Baskin, Carolyn R Bertozzi, David A Tirrell
Affiliations

Live-cell imaging of cellular proteins by a strain-promoted azide-alkyne cycloaddition

Kimberly E Beatty et al. Chembiochem. .
No abstract available

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Figures

Figure 1
Figure 1
Confocal fluorescence imaging of Rat-1 fibroblasts grown for 4 h in medium containing 1 mm Met (first column), 1 m M Aha pretreated with the protein synthesis inhibitor anisomycin (aniso; second column), or 1 m M Aha (last two columns). After the Aha pulse, cells were dye-labeled for 10 min with 50 μM 1 (row 1), 50 μM 2 (row 2), or 10 μM 3 (row 3). Cells were counterstained with MitoTracker Red before imaging. The images for each dye (1, 2, or 3) were acquired under identical conditions to capture either coumarin or MitoTracker Red fluorescence [fourth column, Mito(Aha)]. The scale bar represents 20 μm. Projection images and individual slices from each stack are available in the Supporting Information (Figures S1 and S2).
Figure 2
Figure 2
Flow cytometric analysis of coumarin fluorescence as a function of dye concentration for cells pulse-labeled for 4 h with Aha or Met. A) Mean fluorescence enhancement for cells dye-labeled for 10 min (1 gray bars, 2 hashed bars, and 3 black bars). B) Mean fluorescence values for cells dye-labeled for 10 min with either 10 or 50 μM of each dye. For each sample, 20 000 events were collected.
Figure 3
Figure 3
Histograms of coumarin fluorescence from flow cytometry. Cells were pulsed for 4 h in media supplemented with 1 m M Met (red), 1 m M Aha +anisomycin (aniso; black), or 1 m M Aha (blue). After the pulse, cells were dye-labeled for 10 min with 50 μM 1, 50 μM 2, or 10 μM 3. Values given indicate the mean fluorescence for each population of cells.
See this image and copyright information in PMC

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