Has anyone used enzymatic digestion to isolate myeloid cells from human breast tumors? If yes, which enzymes and could you please share the details?

I need to isolated myeloid lineage cells from human breast cancer tumors. Downstream applications would be tissue culture, flow cytometry and functional assays. In the lab we use enzymes Liberase...

09 October 2016 6,122 3 View

I need to label plated macrophages with CFSE? Has anyone does this before?

10 November 2015 1,535 4 View

How do I differentiate monocytes into a phenotype similar to Tumor-associated macrophages (for breast cancer)?

Would a good approach be to use supernatants from various breast cancer cell lines and differentiate these CD14+ monocytes from healthy donor blood into macrophages in their presence for 10-14...

01 January 1970 1,975 2 View

How much pressure we can reduce at the outlet to avoid Reverse Flow?

I am simulating Heat Exchanger. I have tried and tested all the methods to resolve this issue from the internet like refining mesh, improving skewness and orthogonal quality ,extending outlet to...

01 March 2021 6,985 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

What test should I use to compare two or three means, if data is not normally distributed but sample size is > 30 in each group?

I personally feel this as a most confusing question. As per my understanding, if sample size is > 30, and sample is skewed, we can still use paramteric approach, as it will follow Z...

01 March 2021 1,587 6 View

How to increase mesh quality of a cfd model in ansys fluent?

How can one improve mesh skewness, orthogonality, aspect ratio?

28 February 2021 7,730 3 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

How to find a correlation between coordinates in a curvilinear coordinate system?

Hello. I have a skewed elliptical coordinate system (I use alternative elliptical coordinates). And I need to determine connections between coordinates in order to fetch a covariance matrix of...

28 February 2021 364 3 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View