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Migrated cells on the bottom surface were stained with Coomassie blue.
Migrating cells on the lower membrane surface were stained with hematoxylin and eosin, and counted under an optical microscope at 40x.
Migrated cells remaining on the bottom surface were stained for AQP1 for wild type cells and counted 6hr later (see immunofluorescence for AQP1).
Confocal microscopic images shown in Figure 4b indicated that viruses expressing PS11-scFvFc-CD28-gp41 (706–713) on their surface were stained with both anti-HIV-1 p24 antibody (image a) and anti-human Fc antibody (image b).
Cells on the lower surface were stained with Giemsa after being fixed with 5% formalin.
Migrated cells remaining at the bottom surface were stained with Crystal violet.
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Visual perceptibility of changes in image is not only related to variation but also to roughness of the image, just like when a smooth surface is stained, it is easy to identify but when the surface is rough, it is difficult to identify the stain.
The cell surface was stained and then permeabilized with Cytofix/Cytoperm solution (BD PharMingen).
The cell surface was stained with α-GH and a fluorescently labeled secondary antibody, or with WGA-488 from Life Technologies.
Each well surface was stained by adding 100 μL of 0.3% (w/v) crystal violet (Merck) in water and kept for 5 min.
The sections were mounted on plastic slides and ground to approximately 50 μm in thickness, and the surface was stained using a Giemsa stain modified for plastic sections [ 31].
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