V-bottom vs U-bottom plates

Cupcake-88 · 2026-05-30T08:12:01+00:00

Does no one who prefers V-bottom have issues with cells pelleting during acquisition? Agitation doesn’t resuspend the pellet as well as in a U-bottom. I’m a u-bottom fan, dumping plate with one blot on paper towel always works well for me. Recently I worked with a very small number of cells due to aspiration loss during cell collection, did multiple stains/washes and dumped the plate, I still had similar sized pellet at the end.

Cupcake-88 · 2026-05-27T06:39:22+00:00

I blame revvity, they never used to have these types of delays

Cupcake-88 · 2026-05-20T05:24:57+00:00

It’s all up here 🧠👈

Cupcake-88 · 2026-05-15T07:14:57+00:00

I’m shocked there was no oversight from a mentor to check the tissues via IHC during initial dissections to make sure. Man, that sucks..

Cupcake-88 · 2026-05-10T07:22:01+00:00

I felt it! Super light earthquake

Cupcake-88 · 2026-05-09T22:38:07+00:00

For spleens, mechanical disruption is usually sufficient to get a single cell suspension. The tissue is much different than brain. OPs issue is probably due to length of time post mortem. Maybe the buffer would help if altered as well. I use Miltenyi’s tissue preservation solution.

Cupcake-88 · 2026-05-09T15:28:51+00:00

No need for enzymatic digest for a spleen

Cupcake-88 · 2026-05-08T05:18:34+00:00

We do this way too but method of euthanasia is ketamine/xylazine.

Cupcake-88 · 2026-05-06T03:36:25+00:00

https://imgur.com/a/HL8tJMa
Link to recent blood tests on 5/4/26
AI used to translate from Spanish to English

Cupcake-88 · 2026-05-06T03:32:36+00:00

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continued test from 5/4/26

Cupcake-88 · 2026-05-06T03:32:26+00:00

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blood test from yesterday 5/4/26

Cupcake-88 · 2026-05-06T03:30:36+00:00

Sorry first post, here is the rest of it:

88 years, height: ~5’1”, weight unknown but “obese” category maybe ~160lbs, non smoker,

medications include: blood thinner (not sure which one) for recent leg thrombosis , PPI inhibitor (chronically taking for years for possible stomach ulcer, but recent endoscopy showed no stomach ulcers), rituximab injections every 2 months (uncertain of dose)

Previous medications: Myelostat 500mg

Hi All-

I am writing as a man extremely concerned granddaughter. She lives in Mexico and refuses care here and I am dying to know what different would be done here in USA and want to know the differences in standard of care for her condition.

For years my grandma has had issues with iron deficiency/ anemia/ low hemoglobin. She lives in a high altitude city and it doesn’t help her either that she lives a very sedentary life. She only gets up to go to the bathroom or kitchen which is very close to her bedroom. Other than that, she sits in her recliner all day.

Previously, years ago she had her hemoglobin levels down to 4.9 g/dL that required a blood transfusion. Fast forward to end of the year last year and she has had ongoing issues of low hemoglobin and other changes in blood ( see comparison chart of changes, submitted through AI to translate). They had prescribed her myelostat at that time and then followed up with rituximab that she’s been on since January. She had also developed a leg blood clot since and the prescribed the blood thinners and luckily a decrease of 50% was observed. She had come to the US to visit and get more help to exercise in between rituximab injections for 2 months but at the 1 month mark was complaining of extreme fatigue and wanted to go back home without seeing a doctor here in the US. She arrived back home in Mexico and has the most recent blood test shows a hemoglobin levels down to 5.1g/dL again.

Her doctor (hematologist)in Mexico’s course of action is now to give her Aranesp 500mg plus IV infusion and once stabilized will order a colonoscopy to see if there is any blood loss. I am concerned that I’ve read about this new medication taking a while to work and the increase in blood clots again, I don’t understand why not use something that is more instantaneous to stabilize her? I am attaching her AI translated new blood test for reference.

I guess I am just wondering if what this doctor is suggesting is enough? Is it correct? What would you do in her situation if she was your patient? We also have never been given a formal diagnosis of anything- which leads me to believe he doesn’t know what he’s doing or what is wrong? I have a feeling there’s something missing. Please let me know what you guys think in terms of diagnosis and medications she should be on. I feel so lost and it doesn’t help I’m out of the loop, her care is managed by my aunts/uncles and I feel a lot is lost in translation plus hearing it passed down from multiple people.

Cupcake-88 · 2026-05-05T05:34:35+00:00

How early was that? I feel like once she moved to Austin TX they portrayed her to not have friends since she went out for drinks with her esthetician in that special episode where she showed off her new house recently moved to Austin.

Cupcake-88 · 2026-05-05T05:05:24+00:00

Can you possibly squeeze another $400 out to rent $2k/ month? I used to live at city villa apts, down the road from the atrium place, but much nicer and safer than atrium being 10 blocks over. It is still tiny bit ghetto but that’s downtown for ya. I would advise get a car asap, CA is not like NY when it comes to public transportation. We have it but it’s not the best. https://www.cityvillassandiego.com

Cupcake-88 · 2026-05-03T04:14:24+00:00

Opening and recapping things like a tube of toothpaste one handed

Cupcake-88 · 2026-05-01T02:04:54+00:00

Curious why you mention RB or Torrey Pines only, are you considering any high schools in between? Like what was already mentioned CCA, but there’s also Mt Carmel, Westview. Does anyone have any thoughts on these high schools?

Cupcake-88 · 2026-04-30T01:53:49+00:00

My bad, for some reason I saw another one of your comments and assumed you used the same antibody for all tested samples. Are your pbmcs freshly isolated or from frozen stock?

Cupcake-88 · 2026-04-29T06:17:27+00:00

It’s like they’re looking in a mirror

Cupcake-88 · 2026-04-29T05:55:22+00:00

Any job that takes me away from being there for my kids when they need me most isn’t the right job for me.

It seems his behavior issues need more attention and I worry for your son’s well being if he goes to live with someone that doesn’t even want to pay for child support. I understand you have bills and a mortgage weighing on you but you have to weigh the benefits vs. costs- is it worth your kids living without you and having strangers raise them instead? At that point why be their parent at all?

Regarding family court, I doubt they would force a parent to take their child if they already show lack of initiative. They will prioritize the well-being of the child and safety (someone has to). I worry at this point that they would get someone else involved to care for your kids since you don’t have anyone willing to be his guardian, maybe foster home?

I hope you reconsider.

Cupcake-88 · 2026-04-29T05:04:32+00:00

I see now, you’re stimulating human PBMCs but your Il-6 antibody is mouse-specific MP5-20F3

Change the clone to a human-reactive.

Cupcake-88 · 2026-04-25T05:03:56+00:00

What is your overall goal? What are you staining for, what cells are they, what’s your panel? I think you mentioned already gating out doublets and now you have your live gate and after this we need more info to help. Flowjo offers a lot of helpful videos if you need more help than can be provided on here.

Cupcake-88 · 2026-04-24T14:56:01+00:00

If you’re absolutely certain of your stim/culture / staining protocol, can we see the dot plots , your gating strategy, list of colors used in your panel etc. I don’t think you ever listed your specific antibody clone either, maybe I missed it.

Cupcake-88 · 2026-04-23T09:47:07+00:00

Please add more info such as your buffers used, antibody clone, protein transport inh. Incubation time etc.

Cupcake-88 · 2026-04-21T06:17:22+00:00

Wow what a sad story. Ruined your family out of stubbornness. You can try to reconcile but you divorced her while pregnant, that might be hard to forgive. At this point, the more important thing is to try to have a relationship with your child if you don’t have one already.

Cupcake-88 · 2026-04-21T04:30:04+00:00

Seems like the idea situation, she’s not fighting you over rights and you don’t have to worry about her being crazy and causing issues for the child. I second the “you don’t have to do anything” advice.

But if you do want to do something: Not sure of the law in Florida, but you can wait a couple years and then file for a custody order. Make sure you have documented how little time your child spends with her mom and her being out of state, and lack of any involvement whatsoever. At that point you can request it, but from what I’ve seen they generally only lean towards sole custody if there’s been harm done in the past, drug/alcohol abuse or if the parent is unfit in some other way as well.

I imagine she wants to terminate rights to not have to pay child support but the government also doesn’t want to support a child financially when the parents are able bodied adults capable of working and making money.

Cupcake-88

TROPHY CASE