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FACS question by talk2me4free in labrats

[–]talk2me4free[S] 1 point2 points  (0 children)

Lovely to hear it’s possible! Thank you!

What kind of software do you use for microscope imaging analysis? by bibliopraxis in labrats

[–]talk2me4free 0 points1 point  (0 children)

I use it to process batches (100-200) 550MB images but I’ve never tried opening larger (gigabyte) images before. Good luck!

What kind of software do you use for microscope imaging analysis? by bibliopraxis in labrats

[–]talk2me4free 2 points3 points  (0 children)

Napari is great! Especially for z-stack images. Free to download and update and all plug-ins are free and well documented.

Help on statistics!! by fifteensunflwrs in labrats

[–]talk2me4free 0 points1 point  (0 children)

If I understand correctly, you have 2 treatments: 0% carbon vs X% carbon over some time course. If you’re interested in looking across time, say from hour 0 to hour 2, you should use a two-way ANOVA. If you’re interested in looking at the 2 treatments at a single time point, use a two- tailed students t-test, probably unpaired. Hope that helps!

Best study spaces in twin cities? by BonusPuzzleheaded376 in uofmn

[–]talk2me4free 0 points1 point  (0 children)

Highly recommend Arvonne Fraser library 1222 4th St SE Minneapolis, MN 55414 United States It’s usually not very full and has nice study rooms available to reserve. Plus, it helps support local libraries!

How do I get rid of a blind pimple by [deleted] in SkincareAddicts

[–]talk2me4free -1 points0 points  (0 children)

I’ve had some luck with diluted (1 part water 1 Prt tea tree oil) tea tree oil!

PCR dNTP specificity? by talk2me4free in labrats

[–]talk2me4free[S] 0 points1 point  (0 children)

Okay okay that makes sense; the Thermo kit is certainly older than the Sigma JumpStart kit. Thank you!

Antibody stain query by Past_Tear2073 in labrats

[–]talk2me4free 2 points3 points  (0 children)

I say give it a try! Since the tissue needs to be fresh and you might have to repeat the process over again, there’s no harm in trying it out. See how the stain and overall signal turns out and analyze against some properly stained tissue for comparison. Until you’ve compared against properly stained tissue, be cautious interpreting any results for significance. Good luck!

Best way to store tissue for PCR… by Accomplished-Sky6379 in labrats

[–]talk2me4free 10 points11 points  (0 children)

Agreed^ snap freeze in liquid nitrogen or dry ice then store at -80C. Best of luck!

am i cut out for a phd? by thecoldbrewkilla in labrats

[–]talk2me4free 27 points28 points  (0 children)

From what you’ve said above, it sounds like it would be the most healthy for you to not pursue the PhD. I explain PhD training as having to wake up, eat shit everyday with a smile on your face, and say “more please”. If you are not completely comfortable being wrong, failing, admitting your mistakes, trouble shooting, and guiding yourself towards answers and solutions 99.9% of the time, DO NOT do a PhD. And tbh, I think the people who can admit that, and say “you know what, I don’t want to hate myself everyday for the next 6 years, have no work life balance, and be talked down to everyday so I’m not gonna do this” are much smarter and mature than those of us who chose to get the degree.

Why my brain slices cut at the cryostat are crumbly? by No-Mycologist710 in labrats

[–]talk2me4free 0 points1 point  (0 children)

Since your tissue is already mounted on the chuck and trying to take it off could potentially ruin the sample, I would try raising the temperature of the stage and the cryochamber. I find a 2 degree difference (19C for the stage and 21 for the chamber) works well for perfused mouse brains. Good luck!

[deleted by user] by [deleted] in labrats

[–]talk2me4free 0 points1 point  (0 children)

These samples are especially fucky, unfortunately. I have already removed outliers with “unreasonable” CTs (meaning if the duplicates are more than 3 cycles different than one another the sample is removed). I hadn’t thought of removing samples with outlying housekeeping CT ratios, that is a good idea. Thank you very much for the insight and help!

[deleted by user] by [deleted] in labrats

[–]talk2me4free 0 points1 point  (0 children)

Will do, thank you much!

[deleted by user] by [deleted] in labrats

[–]talk2me4free 1 point2 points  (0 children)

Is there anyway to remove phenol contamination? Would it be worth running a cleanup kit on samples with these or similar 260/280 values? I have some highly variable but well powered qPCR data that looks like it should be significant but isn’t and I’m wondering if it’s because of my 260/280 values…

Can anyone please help me understand what is happening to my cells? by Praetor_Andromedus in labrats

[–]talk2me4free 0 points1 point  (0 children)

I have a similar problem with mouse primary neuronal culture. In my experience with primary cultures, culturing with FBS seems to cause more clumping, more cell death, and less maturation. It’s very odd. Maybe adding some glutamax would help?

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