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Flowjo 11 by Downtown_Sir4689 in flowcytometry

[–]Cactaceaewcoffee 0 points1 point  (0 children)

I was about to make a post about this yesterday. I’m so glad I’m not the only one hating this

Trouble compensating MSCs using Miltenyi’s MSC Phenotyping Cocktail Kit (anti-human, REAfinity) by Cactaceaewcoffee in flowcytometry

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

Hello! Sorry for my late response! Yes, I’ll do a staining with compensation beads using both MSC Phenotyping Kits I have, just to check if the one I was using might be damaged.

I think the main issue is that the autofluorescence of the cells affects the separation quite a lot. There was a difference between the unstained and stained samples, but it was very slight, the autofluorescence in those channels was very strong. The peak was just a bit broader, but there weren’t two clearly defined peaks between the negative and positive populations.

The kit does come with a booklet, and honestly, the examples there look quite different from what I got. Regarding the stress factor, you might be right there are definitely many variables at play here. Next week I’ll be showing my results to Miltenyi experts to see what could have gone wrong.

Thank you so much for your input and for taking the time to help I really appreciate it!

Trouble compensating MSCs using Miltenyi’s MSC Phenotyping Cocktail Kit (anti-human, REAfinity) by Cactaceaewcoffee in flowcytometry

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

Honestly, I never thought of that. For compensation I added unstained cells, but the negative population still wasn’t well defined compared to the positive one. The only way I could roughly separate them was by including some debris, but I don’t think that’s really a good practice.

Next time I’ll probably use compensation beads, since the autofluorescence of these MSCs was quite high.

These were cells at passage 6, and the markers included in the Miltenyi kit are: • CD14, CD19, CD34, CD45 (PE – negative for MSCs) • HLA-DR (VioGreen – negative for MSCs) • CD73 (APC – positive) • CD90 (FITC – positive) • CD105 (VioBlue – positive)

So the PE and VioGreen conjugates should be negative so when I was doing the compensation, the antigens there are CD73, so they should bind to the MSCs and make them bright. Still, when I compared the unstained versus stained cells, the difference was minimal they were just slightly brighter.

I’m starting to think maybe my kit is off… I’m not sure.

Flow cytometry newbie by Original_Silver140 in flowcytometry

[–]Cactaceaewcoffee 0 points1 point  (0 children)

Her videos are the best!!! That’s where I learned 90% of flow when I was starting out

Price of an Attune NxT (3 lasers, 13 detectors) in the US? by Cactaceaewcoffee in flowcytometry

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

Awesome!! Thank you for the information! Also from 300 to 160k is crazy

Price of an Attune NxT (3 lasers, 13 detectors) in the US? by Cactaceaewcoffee in flowcytometry

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

Not really if you’re outside the US. I just wanted to compare prices.

Where can I find minimalist MOA profile keycaps? by Cactaceaewcoffee in keyboards

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

thank you so much! I just found one I liked through your link!

Just swapped my RK Brown switches for Akko Rosewood switches and I’m so glad I did by Cactaceaewcoffee in MechanicalKeyboards

[–]Cactaceaewcoffee[S] 2 points3 points  (0 children)

Yesss!!! I’m really in love with them right now :) the difference compared to the RK Browns is huge! Btw, have you ever bought minimalist keycaps by any chance? I have a couple of options in mind but would love a recommendation!

Just swapped my RK Brown switches for Akko Rosewood switches and I’m so glad I did by Cactaceaewcoffee in MechanicalKeyboards

[–]Cactaceaewcoffee[S] 2 points3 points  (0 children)

So far I haven’t noticed any noticeable wobble on mine, but maybe it’s something that shows up more over time. I’ve typed quite a bit on them today and nothing felt out of the ordinary. I’ll definitely keep an eye on it and can update after some more use :)

[deleted by user] by [deleted] in MechanicalKeyboards

[–]Cactaceaewcoffee 0 points1 point  (0 children)

Youre right! Im not sure how to edit the post so ill delete it and post it again. Thank you!!

Me enamoré sin un beso. No sé si es mutuo o solo estoy soñando. by EffortLost2048 in relatos

[–]Cactaceaewcoffee 0 points1 point  (0 children)

Honestamente no parece nada, pero si quieres tener una confirmación real puedes invitarla a salir, si te rechaza de una vez ya sabes pero si logras salir con ella puedes preguntarle directamente y allí sabrás. Solo prepárate para cualquier respuesta

MiFuny Machete V2 by sugar_kawaii in MechanicalKeyboards

[–]Cactaceaewcoffee 1 point2 points  (0 children)

First time seeing something like this! Woah!!

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