live-cell microscope nikon eclipse ti-e  (Nikon)

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Amino-Terminal Part of the Needle-Tip Translocator LcrV of Yersinia pseudotuberculosis Is Required for Early Targeting of YopH and In vivo Virulence

doi: 10.3389/fcimb.2016.00175

Figure Lengend Snippet: Pore forming activity does not correlate to translocation ability. (A) Intracellular localization of YopE in HeLa cells after 1.5 h infection was analyzed with immunostaining. The samples were viewed using laser scanning confocal microscopy and green staining corresponds to YopE, while the HeLa cell membranes are stained in red. (B) To analyze translocation of YopH, Hela cells were labeled with the FRET substrate CCF4-AM and infected with the indicated strains expressing YopH-Beta-lactamase fusion protein (YopH-Bla). Images were acquired in a live cell microscope using a longpass filter. Translocation of the Bla-fusion results in a shift of fluorescence from green to blue. Both experiments were repeated at least three times.

Article Snippet: Images were taken with a live cell microscope (Nikon Eclipse Ti-E), equipped with a true color camera, using a long pass filter to detect the two wave length of the FRET substrate.

Techniques: Activity Assay, Translocation Assay, Infection, Immunostaining, Confocal Microscopy, Staining, Labeling, Expressing, Microscopy, Fluorescence

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Amino-Terminal Part of the Needle-Tip Translocator LcrV of Yersinia pseudotuberculosis Is Required for Early Targeting of YopH and In vivo Virulence

doi: 10.3389/fcimb.2016.00175

Figure Lengend Snippet: The LcrV N-terminus is required for functional phagocytosis inhibition . J774.a1 macrophages were infected with Y. pseudotuberculosis for 30 min after which the cells were fixed and extracellular as well as total bacteria were detected using a double staining technique. For each sample, the amount of total and extracellular bacteria of 50 infected cells was counted manually using a fluorescence microscope. The results presented are the mean value ± SEM of five independent experiments. The result of each experiment was analyzed using the Wilcoxon signed-rank test and the significance was set at p ≤ 0.01 ** and p ≤ 0.001 *** .

Article Snippet: Images were taken with a live cell microscope (Nikon Eclipse Ti-E), equipped with a true color camera, using a long pass filter to detect the two wave length of the FRET substrate.

Techniques: Functional Assay, Inhibition, Infection, Bacteria, Double Staining, Fluorescence, Microscopy