When Something Finally Worked

It happened! Our trial finally worked! After a few weeks straight of straight troubleshooting, we have finally managed to pull off a successful trial! Over the past few weeks we have dealt with problems with cell count. Before plating, we have to use a hemocytometer to count cells so that we can control how many cells we put into each well. When we first started conducting the trials on our own, our counts were so low, making it nearly impossible to start trials. We started to become more careful when culturing flasks and becoming more selective with which flasks to use. We also encountered issues with the fixation process throughout our trials rendering all of our cells dead on our slides. This was a problem that left us at a loss because we would see cells on our slides before we started fixation, but not afterwards. We figured out that when we were aspirating the washes, we were leaving our cells in too dry of an environment, so we began to increase our speed when fixing our cells. We also decreased the amount of washes so that we were not disturbing our cells too much. After our fixation and mounting process, we looked at our cells underneath a microscope and for our previous trials, there were very little neurites on the cells. This was concerning because we stimulated our cells with the proper neural growth factors that should cause more neurite extensions to grow. We ended up increasing the concentration of neural growth factor to help stimulate the growth. After all these changes made to our procedure, we have finally seen a successful trial! The cells look beautiful, healthy, and happy underneath he microscope with plenty of neurite growth. It has been very exciting to see the fruits of our labor finally come into fruition. I look forward to using these experimental techniques on other cell lines.

Comments

  1. kmreed01 says:

    Hi Ashley! I’m glad that you were able to optimize your experimental procedures, it sounds like those setbacks were quite frustrating. I’ve also been having some trouble with cells dying during fixation, so I completely understand what it’s like to work so hard on trials only to not see cells on the microscope slide. What other cell lines do you plan on working with? Perhaps actual neurons to compare to the PC12 cells, which are only neuronal models? Good luck with the rest of your summer, I hope your trials continue to be successful!

  2. I am so happy for you!! I’m glad that you are finally seeing the fruits of your labor. Patience and hard work is definitely providing you with good returns now. I hope that you will be able to see more successful trials with more neurite growths soon. I can’t wait to see more of your results!