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Questions related from Evan Chiswick
I want to label PA with several fluorophores to assay phagocytosis and phagosomal processes. Unfortunately the bacteria aggregate during the labeling process (both live and heat killed @ 80C...
12 December 2015 2,433 4 View
Luminex, and several publications, report that carboxylated polystyrene beads should be activated with EDC:Sulfo-NHS in pH 6.2 phosphate buffer, and then coupled with antibody in pH 5.0 MES...
02 February 2015 4,324 14 View
If mice are given one single dose (0.5mg) of anti-Ly6G (1a8) IP to deplete neutrophils, how long will it take for the neutrophils to be replenished to near normal levels?
01 January 2015 8,104 7 View
I plan to sort peripheral blood neutrophils from septic mice to determine differential gene expression between groups. The sorting will be done on a BD ARIA II flow sorter. How are cells typically...
06 June 2014 1,042 4 View
I want to sort macrophages/neutrophils from septic mice based on whether they phagocytize bacteria, generate ROS, and/or acidify their phagosomes. Then, I would look for difference in...
11 November 2013 8,394 1 View
I am using an LSRII w/ FACS DIVA software, 405/20bp for Indo-violet and 525/50 505lp for indo-blue. What should I have the scaling factor set to for the Ratio of Blue/Violet fluorescence? Is...
07 July 2013 7,499 3 View
Does anyone know of any differences between the HK1.4 and the Al-21 clones, as far their ability to distinguish cell populations by Flow cytometry. I understand they are of different isotypes, but...
06 June 2013 4,337 3 View
In setting the gates for positive events via Fluorescence Minus One (FMO) controls, do you include an isotype matched antibody in the FMO cocktail, or leave that channel empty altogether? I.e.:...
03 March 2013 2,716 16 View
I am looking at peritoneal macrophages and neutrophils in mice that have extreme peritonitis. The non-survivors have decreased phagocytosis (pHrodo e.coli) and reactive oxygen burst (DHR-123, stim...
03 March 2013 6,582 14 View
I want to determine how many granulocytes infiltrate the kidneys in response to a toxin. How do I quantify them with absolute cell counts? My method briefly: Digest kidney with collagenase &...
02 February 2013 4,618 6 View
I am trying to isolate RNA from peritoneal leukocytes 6 hours after induction of acute peritonitis (cecal ligation and puncture.) I lavage the belly with hbss/edta and some of my samples are...
01 January 2013 6,323 12 View
I want to use a gene array to determine differential gene expression of leukocytes in sepsis, but the bioanalyzer results all show significant bacterial RNA contamination. My model induces sepsis...
07 July 2012 7,684 2 View
