Cells grown on stroma on Lab-Tek® Chamber Slide (Electron Microscopy Sciences) were fixed with 3% paraformaldehyde for 20 min at room temperature and then permeabilized with phosphate-buffered saline (PBS) plus 0.1% Tween-20 for 45 min. After blocking with PBS containing 2% bovine serum albumin (BSA) for 10 min, the cells were stained with anti-phospho FAK antibody for 45 min, followed by TRITC-conjugated secondary antibody for 30 min. The slides were treated with Vectashield containing DAPI (Vector Laboratories) and mounted. Images were collected using Nikon A1SiR Confocal Microscope and processed using NIS element confocal imaging software.
Lab tek chamber slide
Manufactured by Electron Microscopy Sciences
Sourced in United States
The Lab-Tek® Chamber Slide is a specialized laboratory product designed for cell culture applications. It consists of a glass microscope slide with a removable plastic chamber or well attached to the surface. The chamber provides a contained environment for cells to grow and be observed under a microscope.
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Lab products found in correlation
3 protocols using lab tek chamber slide
1
Immunofluorescent Staining of Phospho-FAK
2
Immunofluorescent Staining of Phospho-FAK
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Cells grown on stroma on Lab-Tek® Chamber Slide (Electron Microscopy Sciences) were fixed with 3% paraformaldehyde for 20 min at room temperature and then permeabilized with phosphate-buffered saline (PBS) plus 0.1% Tween-20 for 45 min. After blocking with PBS containing 2% bovine serum albumin (BSA) for 10 min, the cells were stained with anti-phospho FAK antibody for 45 min, followed by TRITC-conjugated secondary antibody for 30 min. The slides were treated with Vectashield containing DAPI (Vector Laboratories) and mounted. Images were collected using Nikon A1SiR Confocal Microscope and processed using NIS element confocal imaging software.
3
Cell Invasion Assay with Geltrex Coating
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The cell invasion assay was modified from Bayless et al. [60 (link)]. Briefly, a Secure-Seal® Hybridization Chamber Gasket (Thermo Fisher Scientific) was adhered to the well of a glass Lab-Tek® Chamber Slide™ (Electron Microscopy Sciences; Hatfield, PA, USA). Subsequently 20 μL of Geltrex® was added through one port on the Secure-Seal® coverslip. The slides were incubated for 30 min at 37 °C for gel solidification. 5 × 104 cells were then added through the other port on the Secure-Seal® chamber and the samples were incubated for 5 h in the presence or absence of rmMCP-7. After incubation the samples were fixed with 2% paraformaldehyde for 20 min and washed in PBS. The slides were analyzed using a Leica DMI600B9 microscope (Leica Systems, Wetzlar, Germany) and the images were acquired using a Leica DFC365FX (Leica) digital camera. The distance traveled by the cells was analyzed using the ImageJ program [61 (link)].
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