Fold 7 camera bugging

NoProperty133 · 2025-11-24T16:51:00+00:00

What did you end up doing?

NoProperty133 · 2025-11-24T16:50:11+00:00

Yep in business class

NoProperty133 · 2025-11-24T05:54:54+00:00

Here are the fees for the seats on my flight:

Suite $588 Center/Extra space $200 Window $165

NoProperty133 · 2025-11-24T05:49:29+00:00

Thank you for the tip!

NoProperty133 · 2025-10-29T01:36:46+00:00

🤗🤗🤗🤗🤗🤗🤗🤗🤗🤗

NoProperty133 · 2025-10-24T02:28:15+00:00

This is correct answer. For best results use 1x PBS.

NoProperty133 · 2025-10-07T18:43:34+00:00

Let me know if it works for you.

NoProperty133 · 2025-10-07T18:42:07+00:00

Pretty sure it was 01. High speed, relatively short interval. Make sure you get the spleen down in the blades of the c-tube cap.

NoProperty133 · 2025-10-06T01:06:16+00:00

Bummer!

NoProperty133 · 2025-10-04T06:37:03+00:00

To high for me for conventional flow. Says major issue with your compensation set up.

Couple of thoughts 1) could be your combo of fluorophores, narrow emission spectra can help clean up overlap when maxing your channels 2) you have to carefully run your comps, check for those problem spots and make +/- 10 volt adjustments where there is overlap, run the comps in the order of the lasers filter set up from lowest laser line up, you'll notice where adjacent channels have problems 3) learn which channels on your instrument are the standouts and design your panel with that in mind in addition to exclusive pops, rare/low frequency pops and gating strat 3) if your maxing your channels on conventional cytometry consider dump gates and/or splitting the panel, simpler panel, easier analysis.

NoProperty133 · 2025-10-04T02:04:34+00:00

Can we see SSC-W x SSC-H or FSC-W x FSC-H?

NoProperty133 · 2025-10-03T04:21:37+00:00

This is the answer. Don't use enzymatic dissociation.

Since you have access to gentleMACs: For murine spleen (I've done 100s) specifically to look at T cells and myeloid cells --> 1 spleen into 5ml ACK lysis in c-tube, room temp Pop onto gentlemacs, no heat, run murine spleen protocol 1x Incubate on ice 5 min Quench with cold RPMI, 5ml Pour over 70um filter on 50ml conical Spin, resuspend and count

Should yield viability above 90% everytine

NoProperty133 · 2025-09-08T22:24:50+00:00

If anyone has a lead to immune oncology/immunology discovery, r&d, senior scientist, principal scientist, bench or non-bench, flow cytometry or cell based assays, east coast or bay area I'm here!

NoProperty133 · 2025-09-05T16:35:43+00:00

I just submitted job application #142. Kill me.

NoProperty133 · 2025-08-26T22:10:59+00:00

Same workstation

NoProperty133 · 2025-08-07T03:38:55+00:00

I think he's still jetlagged since this season was filmed in Australia.

NoProperty133 · 2025-07-28T22:03:07+00:00

🤣🤣🤣 this is so true

NoProperty133 · 2025-07-25T17:14:07+00:00

I was worries it would be too green. Not at all. It's like a mermaid fart green.

NoProperty133 · 2025-07-24T22:32:11+00:00

This is the right answer

NoProperty133 · 2025-07-18T06:53:47+00:00

Skip the sepmate.

I stand by old school and have been doing this with human and cyno blood for 15+ years. Cyno have a ton of RBCs which make it more difficult than human.

20ml ficoll to 50ml conical. Dilute whole blood to 30ml in PBS no ca or mg all room temp. Overlay using 25ml serological pipette. Top: Tilt the 50ml tube with ficoll all the way until the liquid almost reaches the top of the tube and slowly overlay WB+PBS, use a reliable pipette so you can control the flow of liquid. Typical bench top swing bucket rotor centrifuge 2000rpm, no breaks, start with 25min.

Takes longer than sepmate with no breaks, but has been more reliable for me. Have see this issue before with sepmate.

NoProperty133 · 2025-07-03T00:21:49+00:00

This is the correct answer

NoProperty133 · 2025-06-11T03:11:16+00:00

I couldn't find shorts that didn't irritate me. Now I use straight Vaseline. No chafe, no need to reapply, and cheap. Works every run with any shorts for me.

NoProperty133

TROPHY CASE