Time Lapse Attempt #1

Today, Dr. Keese, Dr. Renken and I set up the equipment required for our time lapse imaging of experiment. In this experiment we will be examining how BSC-1 and MDCK cells interact with one another when co-cultred. As we have seen in the past, MDCK cells have the tendency to form like-cell islands amongst BSC-1 cells. We would like to see how these islands are formed over time. The basic mechanisms required for our time-lapse experiment are:

• Phase contrast microscope
• special ECIS loading dock with windows, water channel system, and gas inlet
• BSC-1 and MDCK cells
• CO2 pump and bubbler
• Nixon camera with microscope lens attachment
• Water pump 

Here is an image of the time lapse setup:

Most of the equipment used in this setup is to keep the cells healthy & happy. Cells like to stay in a warm, moist, environment- hence the water pump that sends 37 degree Celsius water through the channels of the dock they are encased in. The CO2 pump keeps the cells at ~ 5% CO2. The rest of the setup is pretty self explanatory: the camera is set up to take 1,000 frames over a 72 hour period, and the microscope is for magnifying the cells using phase contrast. Phase contrast is essentially a system of lenses that are arranged to allow a specific amount of light in, thus highlighting cells which are otherwise transparent bodies. Here is a picture that demonstrates how phase contrast highlights a cell:

Cells magnified with traditional bright field microscope (left) and phase contrast (right).

In Friday's experiment, we are just looking a BSC-1 cells using time lapse. This test run will help us work out any kinks such as adjusting the magnification, CO2 levels, etc. in order to be prepared for this Friday when we place a co-cultured sample of BSC-1 cells and MDCK cells into this contraption! I'm not sure if we are going to add ECIS measurements to this Friday's run, although that is possible for us to do. I'm hoping our experiment went well and I'll be able to post a time-lapse movie on my next blog post!