Selected article for: "confocal microscope scanning laser and Imaris software"
Author: Warner, Nikole L.; Jokinen, Jenny D.; Beier, Juliane I.; Sokoloski, Kevin J.; Lukashevich, Igor S.
Title: Mammarenaviral Infection Is Dependent on Directional Exposure to and Release from Polarized Intestinal Epithelia
  • Document date: 2018_2_10
    • ID: 1t8jmunt_9
    Snippet: Caco-2 cells were grown apically on 12-well, 0.45 µM Transwell inserts (Corning) for 21 Days until a polarized monolayer was formed. Cells were then fixed with ice-cold methanol for 10 min at −20 • C, washed and stained on both the apical and basolateral sides of the inserts with antibodies against ZO-3 using monoclonal antibody against zonal occludin-3 (Cell Signaling, cat. # 3704, Danvers, MA, USA) at a 1:1600 dilution. Alpha-dystroglycan .....
    Document: Caco-2 cells were grown apically on 12-well, 0.45 µM Transwell inserts (Corning) for 21 Days until a polarized monolayer was formed. Cells were then fixed with ice-cold methanol for 10 min at −20 • C, washed and stained on both the apical and basolateral sides of the inserts with antibodies against ZO-3 using monoclonal antibody against zonal occludin-3 (Cell Signaling, cat. # 3704, Danvers, MA, USA) at a 1:1600 dilution. Alpha-dystroglycan (α-DG) antibody clone 11H6C4, recognizing fully glycosylated α-DG, was used at a 1:100 dilution (Milipore, cat. # 05-593, Billerica, MA, USA). Hoescht 33342 (Thermo Fisher Scientific) was used for nucleus staining at 1:10,000 dilution was added for 10 min. Transwell filters were cut out and placed on microscope slides, followed by 10 uL of ProLong Gold (Life Technologies), and a cover slip placed on top. Slides were analyzed using an Olympus FV1000 laser scanning confocal microscope and analysis was done using IMARIS software (Bitplane, Version 7.7.1, Zurich, Switzerland).

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