Question

Short Answer 7

You are interested in comparing the epithelium of lung tissue in infected mice vs non-infected mice. Describe two experiments you would design to look at the various cell attachments between epithelial cells and their extracellular matrix.

Answer 1

In order to compare the cell attachemnt (between epithelial cells and their extracellular matrix) in lung tissue of infected mice Vs non-infected mice, there are following two experiments can be designed

1. Cell attachment assays - The lung epithelium of infected mice and non-infected mice can be grown on the coverslips coated with ECM and can be fixed upon atatining full confluency.Following to that the immunocytochemistry can be performed using the fluorophore conjugated phalloidin which labels the F-actin and anti vinculin antibody to visualize the focal adehsion ,then confocal microscopy can be performed to visualize the cell attachment between epithelial cells and their extracelluallar matrix.

2. Wash assays -The lung epithelium of infected mice and non-infected mice can be grown on the ECM-coated cantilever inside the microwell plate chamber.Then, the static cell culture can be washed and the fraction of cells remains adhered to the ECM can be determined.The cell remained attached to the ECM can be further labeled with the integrin antibody (attaches the cell cytoskeleton to ECM), and the cell attachment can be analysed.