Any suggested tricks or protocols to lift off primary human macrophages from well plates without scraping?

Sharon Lee @Sharon_Lee25

05 May 2018 0 4K Report

And does scraping activate cells?/ damage surface proteins on cells?

Badges
Science topic

More Sharon Lee's questions See All

Does company brand of cytokines affect differentiation and/or (M0/M1/M2a) polarization of TH1-P cell line into macrophages?

Concerned that there will be functional differences between products from Peprotech vs. Mitenyi vs. R&D.

05 June 2018 7,386 0 View

What does it mean when transfected macrophages increase in size (forward scatter) and granularity (side scatter)?

Cells treated only the transfection agent or the transfection agent with the miRNA construct (meant for transfection), had a higher forward scatter and side scatter compared to cells that were...

04 May 2018 1,161 1 View

Any comments on whether median fluorescence intensity (MFI) or geometric mean is better to evaluate CD marker expression using flow cytometry?

. .

04 May 2018 6,526 0 View

Any suggestions on how else I can label the nuclei of a cancer cell line that has formed a 3D cellular aggregate?

I used NucBlue Live from ThermoFisher (https://www.thermofisher.com/sg/en/home/references/protocols/cell-and-tissue-analysis/protocols/nucblue-live-readyprobes-protocol.html). I found that this...

04 May 2018 3,880 1 View

Any suggestions on a non-toxic nuclear dye for labelling macrophages while the cells are live?

NucBlue LiveReadyProbes by ThermoFisher (https://www.thermofisher.com/sg/en/home/references/protocols/cell-and-tissue-analysis/protocols/nucblue-live-readyprobes-protocol.html) seemed to seriously...

04 May 2018 5,835 3 View

Is it neccessary to use filtered pipette tips when adding lysis solution to lyse cells for downstream RNA work?

Or is the use of filtered tips only crucial from the point of isolating RNA from the lysate? (i.e. after obtaining the lysate)

03 April 2018 9,465 5 View

Any suggestions on how I can inhibit secretion of cytokines by a cancer cell line without killing the cells?

These 'secretion-inhibited' cancer cells would then be co-cultured together with myeloid cells so I am hoping the environment would not be toxic to both cell types.

03 April 2018 5,179 9 View

Can anyone share how they label monocytes with CellTracker Orange(CMRA)?

I use IMDM as staining buffer (is this ok?),but lose 60% cells after staining. Also, any idea/comment on why we should avoid amine-containing or thiol-containing buffers as the staining buffer?

11 December 2017 2,469 0 View

What markers can I use to characterize macrophage subsets (M1 and M2)? And what about different monocyte subsets (e.g. patrolling, etc)?

For immunostaining of cells that are fixed.

06 July 2017 7,570 3 View

Comments on the easiest method to attach antibodies to paper for dry storage, where antibodies can be rehydrated when it is required for use?

05 June 2017 5,436 0 View

Is changing the medium during the experiment overestimate the results?

I am experimenting with cancer and non-cancer cells in a 12-well plate for 4 days with a seeding density 1*10^4/well, however, I noticed that the control group growth rate slows down on D3. Should...

07 August 2024 2,283 2 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View

Can you recommend a 96-well plate for minimal cell adherence to the wells during phagocytosis experiments?

I have been performing short phagocytosis experiments using 96-round bottom, tissue culture treated, well plates (Falcon, 353077). I culture the cells with the phagocytic cargo for 2-4hrs, wash...

04 August 2024 5,228 4 View

Does long-term culture of murine Monocytes result in macrophage differentiation?

Hi, I am isolating monocytes from the bone marrow using the Mouse Monocyte EasySep kit. I want to treat these cells and monitor expression of specific markers over the course of 10 days. I will...

04 August 2024 7,282 2 View

PBMC infection with virus?

In my study, I intend to infect PBMCs with SARS-CoV-2. After that, I will analyze NK cells by flow cytometry to see if their phenotype changes or if they show degranulation. After the infection, I...

01 August 2024 4,403 4 View

Found this object in the cell culture - do you have any suggestions what it might be?

Dear researchers, Today, we found this object in one of the wells of a 96wp culture plate. The plate contained MEC-1 cells and GelTrex. I am quite curious what this is (because it's not cells). Do...

01 August 2024 2,250 2 View

Publishing a checklist?

Does anybody know of entomological Journals without page charges that would publish an extensive checklist (Hong Kong Aculeates wasps) covering 350 species (of which more than half are new to Hong...

01 August 2024 6,134 2 View

How long will macrophages remain in the M1 state after treatment with 100 ng/mL of LPS?

In my experiment, I treated RAW macrophages with 100 ng/mL of LPS. After 24 hours, I collected the supernatant and replaced it with fresh media. I then quantified the cytokine levels of TNF and...

01 August 2024 476 2 View

Human macrophages cell surface markers?

I have a question about human macrophages cell surface markers.I want to stain the unpolarized human macrophages for flow cytometric sorting without permeabilizing them. It is because I need to...

29 July 2024 236 2 View

HEC 1A & HEC1B Cell Lines?

Hi, Kindly guide me that how many cells of HEC1A & HEC1B Cell lines should I seed for Wound healing assay and which plate type is recommended 6, 12 & 24?. Articles suggested mainly 24...

20 July 2024 4,143 2 View