This week, the task that I was given was to conduct an case study experiment with FLT3, a protein receptor that regulates blood cell formation, to see the effect Ikaros ( transcriptional factor) has on FLT3. Since my project will be similar to this and they have already done research on FLT3 with known expected results, conducting this experiment was definitely beneficial to my training! To test it out, I had four samples. All of them were from PDX2 cell lines- patient derived xenografts that model cancer cells from the original patient. My controls: GFP (green fluorescent protein- no Ikaros at all) and GFP-D (no Ikaros, D= doxycycline that activates GFP expression). My other two samples: IK (had Ikaros, but Ikaros cells not activated) and IK-D (Ikaros, D= doxycycline that activates Ikaros expression). By previous research, the expected results/analysis is that Ikaros when present actually suppresses FLT3 causing cell proliferation.
After running a flow on PDX2-IK (my cell line – PDX2 cells with Ikaros), we realized that the cells we had been testing actually didn’t contain any Ikaros at all! The person who had froze this sample (years ago) apparently didn’t freeze it right, so we ended up having to thaw another sample delaying my experiments a bit. This week, I just ended up practicing more flow on my other sample, PDX2-GFP (green fluorescent protein-to track protein activity with no Ikaros), with data that was distorted 🙁 . I haven’t really mentioned Ikaros that much on my posts, so for a refresher Ikaros is a main transcription factor that my lab studies about! Although we did have this delay, I must admit I have gotten a lot better at flow from this experience, since I did flow almost everyday that week (!!!).
This problem probably wouldn’t have occurred if our lab a bit more cleaner (haha). It actually bugs me a lot. I’m not the cleanest myself, but this degree of monstrous atrocity is honestly unbearable (Justin, my colleague, says I’m butthurt). I always have to clean up the bench just for some space, but it is understandable because my other colleagues are busy running off to finish their experiments. There’s always so much to do in lab and there’s never enough time to finish things (In fact, Matt, my other colleague, just did an all-nighter doing lab work!). It’s like a place where space and time don’t exist.
When I’m not working on my project, I’m helping Matt with his bacteria and cleaning the lab, of course. It always gets messy seconds later, and the lab still ends up looking chaotic. The bacteria Matt and I have been trying to grow multiple times, finally grew presently as I am writing this, and finally took in the plasmid Matt wanted. The FLT3 case study was redone and successful presently (week 4). Point is, that research is a messy process in general. But I’ve learned to embrace it. Things don’t always go to plan, but that’s the fun part! Finding ways to make it work.
Everytime I come back from lab I end up feeling gross and tired, yet definitely content.
edit: I’m super tired and just came back from lab so my brain is a bit fried, please let me know if my science-y terms confuse you.
-Wishing you good luck on your projects, Mabel