Results from Trial 1/ Starting Trial 2

At the beginning of this week, I looked at my Trial 1 slides for no treatment/ chloroquine that I made the week before. I examined the twelve chloroquine slides and took pictures of those slides. I need to perform statistics on the slides but I will do that throughout this week. In general, the majority of the cells on the chloroquine slides were not viable and the cells that were alive were scarce. One explanation for this could have been forceful washing during the cell fixation process. This week when I do cell fixation, I will make sure to be careful when washing the cells using the autopipette. I have also done cell culture for Trial 2 of no treatment/chloroquine. I will add my DNA conditions to my cells today and add the autophagy inhibitor drug, chloroquine, to the cells tomorrow.

Comments

  1. ekgergel says:

    Working with something as delicate as cells can be quite difficult, so it’s completely normal for the cells to act up in the beginning! With fixing, some people that I know have found using micropipette tips at the bottom of a regular 10 ml or 5 ml pipette tip can help to be more gentle. In terms of viability, this is one of the most annoying things for scientists! Some times cells die for what seems like no reason but you have to figure out what it is. Is the plasmid toxic? Was the transfection too long? Did the treatment kill the cells? Troubleshooting is a scientists best friend. I’ll be keeping my fingers crossed for you in your future experiments!

  2. tmprioleau says:

    Hello! and you are absolutely right! I figured out that it was not the plamsid that was toxic to the cells or issues with the transfection procedure. Sometimes during cell fixation, the cells can splash out of the 6 well plate. The drug, chloroquine, that I use is highly toxic to the cells when the drug is on the cells for too long.

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