Search found 18 matches

by douglas.linn
Mon Jan 06, 2020 5:10 pm
Forum: Reagents
Topic: IMC on FFPE kidney
Replies: 3
Views: 229

Re: IMC on FFPE kidney

Hi Marieke, First of all, I really enjoyed your paper, excellent work! We have extensively validated a lot of human antibodies using normal tissue TMAs and did not observe anything unusual specific to kidney compared to any other normal tissues. With commercially available TMAs, I've found the spot-...
by douglas.linn
Fri Nov 22, 2019 6:21 pm
Forum: Reagents
Topic: Anyone with experience with Ionpath conjugation kits or antibodies?
Replies: 0
Views: 535

Anyone with experience with Ionpath conjugation kits or antibodies?

I was just curious if anyone else has used Ionpath's conjugation kits or their antibodies? I've only tested a few of their antibodies (HLA-ClassI, CD56, and H3) and they weren't promising, but these aren't clones I have experience with. I also tested their 89Y conjugation and it seemed to work quite...
by douglas.linn
Wed Nov 20, 2019 2:25 pm
Forum: Reagents
Topic: Mouse FFPE on IMC
Replies: 1
Views: 309

Re: Mouse FFPE on IMC

Hi Matt, We've done quite a bit of mouse IMC work with FFPE tissues. We largely abandoned frozen due to inability to get nuclear markers (Iridiums or H3) to work sufficiently while retaining good membrane staining. We have used a regular microwave or the Biogenex microwave with EDTA pH9 AR buffer (a...
by douglas.linn
Wed Nov 06, 2019 9:04 pm
Forum: Careers
Topic: Senior computational biologist positions open at Merck (Boston & Cambridge MA)
Replies: 1
Views: 1115

Senior computational biologist positions open at Merck (Boston & Cambridge MA)

Hi All, Merck has just opened up 2 Senior Computational Biologist positions. The first position would be focused on analyses of highly multiplexed data sets (ex IMC, CyTOF, multiplex IHC, etc) to support Oncology, Neuroscience, Metabolic Disease, and Infectious Disease teams. More details are here: ...
by douglas.linn
Fri Aug 02, 2019 6:50 pm
Forum: General
Topic: Planning/Calculating Runtime & Experimental Details
Replies: 2
Views: 1410

Re: Planning/Calculating Runtime & Experimental Details

In my experience, acquiring at 200Hz takes a little under 2 hrs per 1mm^2
by douglas.linn
Fri Aug 02, 2019 1:54 pm
Forum: General
Topic: length of time after sectioning
Replies: 2
Views: 1076

Re: length of time after sectioning

We try to stick with as fresh as possible but have gotten fine results with slides 6+ months old. My impression is that it depends on how the cut sections are stored, how abundant/stable your epitope is, and how good your antibody is. At the 1st IMC UGM, there was a discussion (David Rimm I believe)...
by douglas.linn
Mon Feb 04, 2019 7:22 pm
Forum: Reagents
Topic: Standardized retrieval conditions?
Replies: 1
Views: 1164

Re: Standardized retrieval conditions?

Hi Santhosh, I haven't tried anything beyond the recommended EDTA buffer for 10 minutes at 95C. I'm using the Biogenex microwave. I have stuck with these conditions mostly because those conditions are similar to what most antibody manufacturers recommend for IHC, at least for the antibodies I'm usin...
by douglas.linn
Tue Jan 29, 2019 9:04 pm
Forum: Reagents
Topic: Detecting mouse monocytes on FFPE?
Replies: 0
Views: 1814

Detecting mouse monocytes on FFPE?

Hi All,
Has anyone has success detecting mouse monocytes using mouse FFPE tissues? Using IHC, I can readily use Ly6C (ER-MP20) or CD115 (sigma polyclonal), but neither of these antibodies adapt well to IMC. Does anyone have any other clones that work in their hands? Thanks!
-Doug
by douglas.linn
Fri Oct 19, 2018 3:17 am
Forum: Data analysis
Topic: Cell population frequencies in HistoCat
Replies: 12
Views: 5258

Re: Cell population frequencies in HistoCat

Does this issue address your question: https://github.com/BodenmillerGroup/histoCAT/issues/11 It works, but I have to manually count the number of rows from each respective ImageID/ROI. This would become quite tedious when there is a lot of clusters across a large set of ROIs, so I was hoping those...