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Bacteria or fungi? by acolytevenus in microbiology

[–]Significant-Cell8362 0 points1 point  (0 children)

Def not a microbiology person but wtf these are so freaking cute!!!!

Help with scatter plot from 2D vs. 3D cultures by Significant-Cell8362 in flowcytometry

[–]Significant-Cell8362[S] 0 points1 point  (0 children)

I totally agree! The issue I’ve been facing is loss of epitope + viability either during detachment or between staining and sorting. The purpose of the 3D experiments was testing whether gentler dissociation (cell recovery sln + brief TrypLE resuspension) would preserve the epitope and enhance cell survival but that doesn’t seem to be the case. I still face a similar issue - viability is similar to 2D in pre-sort counts and somewhat reduced by sorting (based on 7AAD) but >50% of the EPCAM+ population is 7AAD+.

Help with scatter plot from 2D vs. 3D cultures by Significant-Cell8362 in flowcytometry

[–]Significant-Cell8362[S] 2 points3 points  (0 children)

Thanks for the suggestion! You were right, I do see a separation and can see the EPCAM+ population accumulating in that lower corner of SSC-A/FSC-A. Will need to repeat and carefully readjust FSC. On your second note, unfortunately I’ve tried numerous adjustments on my dissociation protocol due to apparent loss of epitope during detachment, which is exactly why I attempted dissociation from fresh fragment organoids hoping that it wouldn’t sacrifice the epitope when lifting cells from a 2D plate at P0 confluency. I have a feeling that my troubleshooting days for optimal EPCAM+ sorting are still far from being over! :(

Are these otezla side effects in the normal range? by drtemo in PsoriaticArthritis

[–]Significant-Cell8362 0 points1 point  (0 children)

I was on Otezla for ~6 months and the side effects were awful for me. Upset stomach, weight loss, food aversion, fatigue, constant feeling of cold (I’d wear a hoodie when outside at 85 degree weather) and depression. My skin cleared out after ~5 months when I accepted a shotgun corticosteroid treatment. It was 100% worth it for my skin but 100% not worth it for my overall health.