Tissue Morphodynamics Labratory: Weeks 2-3

Day 5 Chicken Embryo

My second week of the lab was a little chaotic due to the Princeton bomb scare, which set back  my laboratory safety training which in turn set back my general laboratory training in microdissections. Due to the Princeton bomb scare, I was unable to start microdissections because the chicken eggs had grown past a viable embryonic stage for tissue culture. My laboratory manager told me it would be another week before I would able to start dissections for tissue culture. The second week therefore consisted of more reading and more articles about different topics, such as RhoA pathways and e-cadherins. Through these articles I learned about how pathways regulate tissue in the epithelium as well as the importance of e-cadherins in developmental biology. At the end of the second week I was also given a host of data and images to organize, analyze, and graph.

Lungs of Day 5 Chicken Embryo

At the start of the third week I finally started to learn the multiple techniques and solutions I would need to use in order to analyze chicken lungs under a confocal microscope; however the embryos were still unable to be used for tissue culture. I started by dissecting the lungs out of a day seven chicken lung under a dissecting microscope using two 5mm forceps. In order to extract the embryo from the egg, I took a syringe and poked a hole at the bottom of the chicken egg and removed some fluid in order to lower the yolk height in the egg. Then using a scissor, I cut a semicircle at the top of the egg, removed the shell and scooped the chicken embryo out of the egg and on to a petri dish filled with saline PBS solution. After successfully dissecting four lungs out of chicken embryos I learned how to fix the lungs in preparation of staining. Fixing the lungs is essentially using a solution in order to preserve the tissue of the organ and allow for storing overnight. After fixing, I stained the lungs using LCAM antibodies, which are antibodies that bind to e-cadherins on the epithelium and creates 
contrast between the epithelium and the mesenchymal tissue.  

 

Fixing and Staining of 4 Day 7 Chicken Lungs

Towards the end of the third week I was given more data to analyze, which was mainly measuring the lengths and area of the epithelium using Adobe Photoshop and ImageJ as well as counting the number of terminal end buds of previously stained chicken lungs. I also attended a microfabrications seminar with others in my lab in order to better understand how to amplify signals using biosensors on a plate.