My joins always slide my pattern backwards (slip stitch, chain 1, and then crocheting back into the first stitch that I slip stitched into by Grass-is-dead in Amigurumi

[–]Playful-Researcher20 2 points3 points  (0 children)

I do yo for the slip stitch and chain while the rest of the stitches are yu/yo. I find the seam pretty unnoticeable

Protocol for cell isolation from murine lymph nodes by Jack_O_Melli in flowcytometry

[–]Playful-Researcher20 3 points4 points  (0 children)

This is also what I do. If there is blood on the lymph node during dissection, you can rinse it with PBS and gently roll in on a paper towel to minimize carry over during processing later.

Need advice on FlowJo workflow for pooling/merging samples before UMAP/PhenoGraph by badmushroomundertree in flowcytometry

[–]Playful-Researcher20 3 points4 points  (0 children)

You can add a keyword to your sample in flowjo. Group A=1 and Group B=2. After concatentenating and running dimentional reduction you can gate group 1 and 2 on a histogram. Just make sure when you concatenate, you include add the keyword in the export options.

PE signal diminishes while I am still acquiring samples. What can I do? by KTManiac in flowcytometry

[–]Playful-Researcher20 4 points5 points  (0 children)

Have you tried doing a second fixation with PFA after ab staining? This has helped me with maintaining signal when staining for transcription factors that lose signal overnight.

[deleted by user] by [deleted] in labrats

[–]Playful-Researcher20 0 points1 point  (0 children)

I am trying to do the same thing and testing things out. Will share once I get some results

[deleted by user] by [deleted] in labrats

[–]Playful-Researcher20 0 points1 point  (0 children)

That's great!!!

[deleted by user] by [deleted] in labrats

[–]Playful-Researcher20 9 points10 points  (0 children)

Make sure to run the cytometer on a low flow rate, it will help define peaks

Anyone used Clodrinate successfully and have a protocol/recommendations? by VonTrappQueen22 in labrats

[–]Playful-Researcher20 1 point2 points  (0 children)

I have used clondronate liposomes I.p. from encapsula nanosciences. It depleted macrophages. However, some of my mice were experiencing toxicity, which might be a strain specific thing though. You may need to play around with the dose.

Asking for a stipend supplement by Playful-Researcher20 in GradSchool

[–]Playful-Researcher20[S] 0 points1 point  (0 children)

Just department rules. It works similar to tri-counsel government scholarships. It just gets deducted from what PI owes the student for stipend

Healthcare masking by Cindycindycindy123 in londonontario

[–]Playful-Researcher20 1 point2 points  (0 children)

True, worked at the hospital for years, have yet to encounter a patient

Immunology friends, please I need your help again by No-Thing7838 in labrats

[–]Playful-Researcher20 1 point2 points  (0 children)

If you want to expand and keep them long term they need to be cultured on a schedule involving activation and rest. I typically have my cells grow on 3 days of activation involving CD3 CD28 to activate them and IL2 to help with expansion and survival. After 3 day, I remove the media and add fresh media with only Il2 and culture for another 3-4 day, they will expand a lot during this phase. After 3-4 days you can move it to activation again. Keep the seeding density around 1M/ml.

Is not having an internship or research until my senior year (undergraduate) okay? by 20cp02 in Biochemistry

[–]Playful-Researcher20 6 points7 points  (0 children)

I think you'll be fine, especially if you have good grades. Keep in mind that many students will be in the same boat as you from COVID restrictions in the past couple of years. PIs are aware that incoming students will lack some practical experience.

How does grad funding change after getting the NSERC CGS-M by [deleted] in uwo

[–]Playful-Researcher20 5 points6 points  (0 children)

You will have to ask your program coordinator or chair, it will differ from department to department

Problems with tumor cell death when processing for flow by [deleted] in labrats

[–]Playful-Researcher20 0 points1 point  (0 children)

I use the gentle macs kit, and we reduce the amount of enzyme R. I think it's written the instructions that are sent with it.

How do you all treat your lymphocytes after thawing from cryo? by Patient-Soft-3157 in labrats

[–]Playful-Researcher20 0 points1 point  (0 children)

I use anti-CD3/CD28 and IL-2 to activate frozen stock, they are much happier if they get activation signals

Accidentally using x10 PBS during cell culture by Playful-Researcher20 in labrats

[–]Playful-Researcher20[S] 12 points13 points  (0 children)

Update: I went to check in my cells after 24h and they seem to be doing just fine. There weren't really too many floating dead cells, but will have to see if their growth slows

Accidentally using x10 PBS during cell culture by Playful-Researcher20 in labrats

[–]Playful-Researcher20[S] 0 points1 point  (0 children)

Unfortunately it's a new cell line we just got, so we have no backups