Journal: Nature Communications

Article Title: Whole blood stabilization for the microfluidic isolation and molecular characterization of circulating tumor cells

doi: 10.1038/s41467-017-01705-y

Figure Lengend Snippet: Hypothermic storage of ACD-anticoagulated blood preserves the viability and integrity of diverse cell types. a Representative images of neutrophils in fresh (0 h) and stored (72 h) blood. Storage in RT leads to cell death and release of nuclear materials, whereas storage in 4 °C (cold) preserves cellular integrity and the distinct segmented nuclear morphology. b Imaging flow cytometry for the quantification of viable (calcein+/caspase−) and apoptotic (caspase+) leukocytes as well as surface marker expression. c Percentages of viable and apoptotic granulocytes (CD45+/CD66+), and their expression of CD11b in fresh and stored blood (** p < 0.01; **** p < 0.0001; one-way ANOVA followed by Tukey’s post test). d Surface expression of CD45 and CD66b on agranulocytes (CD45+/CD66b−) and granulocytes. e Representative images of RBCs in fresh and stored blood. Echinocytes are identified by the distinct spiculations as shown in RT-stored blood. f Percentages of echinocytes as a function of storage (**** p < 0.0001; two-way ANOVA followed by Tukey’s post test). c , d , f The platelet stabilization cocktail treatment (tiro-EDTA) does not affect cell viability, activation, marker expression, or echinocyte formation. All scale bars represent 10 μm. Error bars represent SD

Article Snippet: Briefly, whole blood was incubated with biotinylated antibodies to CD45 (clone HI30; Thermo Fisher Scientific), CD16 (clone 3G8; BD Biosciences), and CD66b (clone 80H3; Novus Biologicals), followed by a second incubation with streptavidin-coupled Dynabeads (Invitrogen), and loaded into a pressurized syringe for processing with the CTC-iChip.

Techniques: Imaging, Flow Cytometry, Marker, Expressing, Activation Assay