Journal: Pharmaceutical research

Article Title: The Extracellular Microenvironment Explains Variations in Passive Drug Transport across Different Airway Epithelial Cell Types

doi: 10.1007/s11095-013-1069-5

Figure Lengend Snippet: Confocal 3D image analyses of the mixed cell cultures confirmed TEER values of the monolayer or multilayer architecture of airway epithelial cells. Images were acquired on day 8 under ALI conditions. (a) Mixed cells on the inserts were incubated for 30 min (37°C, 5% CO2) with dye mixtures containing Hoe, MTR, and LTG staining cell nuclei, mitochondria, and lysosomes. Zeiss LSM confocal microscopy was used for the examination with z-axis scanning. Two dimensional images in xy planes show cell nuclei (blue), mitochondria (red), and lysosomes (green) and cell architecture on the insert are shown in yz planes with the arrows indicating “apical medium”, “cell”, and “membrane”. (b) Development of tight junctions in cell-to-cell contacts was examined for the mixed cell cultures on day 8 in ALI condition. Actin filaments in the cells grown on the inserts were stained with Alexa Fluor® 488 phalloidin (green) and cell nuclei stained with Hoe (blue). For the mixed cells (Calu-3/NHBE = 99/1), the vertical arrows represent “apical medium”, “cell layers”, and “porous membrane”. As shown in the yz planes, the mixed cells with more NHBE cells (Calu-3/NHBE = 1/9 or 1/99) formed multilayers with varied thickness of layers. The arrows point to the top cell layer, bottom cell layer and porous membrane support.

Article Snippet: Cell volume (μm 3 ) was calculated by Metamorph in the confocal 3D images and was used as a representative cytometric parameter to evaluate the distribution profiles in the pure and mixed cell populations.

Techniques: Incubation, Staining, Confocal Microscopy, Membrane

Journal: Pharmaceutical research

Article Title: The Extracellular Microenvironment Explains Variations in Passive Drug Transport across Different Airway Epithelial Cell Types

doi: 10.1007/s11095-013-1069-5

Figure Lengend Snippet: The fraction of Calu-3 and NHBE cells in cell populations consisting of pure (a) Calu-3 and NHBE cultures and mixed cell cultures (b, c) were estimated by fitting the distribution of cell volumes in the mixed cell population using a normal mixture statistical model. Cell volume (μm3) was calculated by Metamorph in the confocal 3D images and was used as a representative cytometric parameter to evaluate the distribution profiles in the pure and mixed cell populations. All the mixed cell cultures (Calu-3/NHBE = (b) 99/1, 9/1, 1/1, (c) 1/9, and 1/99) showed bimodal distributions with two distinct cell populations as indicated with the blue arrows (group 1 and 2). For the multilayers in the mixed cell cultures with more NHBE cells (Calu-3/NHBE= 1/9 or 1/99), the bottom cell layers and top cells were separately analyzed. As shown in (c), bottom cell layers consisted of two cell populations while top cells showed unimodal distribution, reflecting one cell-type in the top layer.

Article Snippet: Cell volume (μm 3 ) was calculated by Metamorph in the confocal 3D images and was used as a representative cytometric parameter to evaluate the distribution profiles in the pure and mixed cell populations.

Techniques:

Journal: The Journal of Cell Biology

Article Title: Motile Properties of Vimentin Intermediate Filament Networks in Living Cells

doi:

Figure Lengend Snippet: Fluorescence intensity measurements along a photobleached GFP-vimentin fibril. Gray-scale pixel values were determined along a bleached vimentin fibril every 2 min with the Metamorph image analysis program. In this case, complete recovery took place in 14 min with a recovery half-time (t 1/2 ) of ∼6 min.

Article Snippet: Position, length, and intensity measurements were made on digitized confocal images using Metamorph image analysis software.

Techniques: Fluorescence