Journal: Transfusion medicine (Oxford, England)

Article Title: Extracellular vesicles in ageing cold-stored whole blood may not compensate for the decreasing haemostatic function in vitro.

doi: 10.1111/tme.13122

Figure Lengend Snippet: FIGURE 1 Used blood products, sampling, sample preparation and downstream analysis. Day 1: platelets and all CSWB and RBC units were donated by the blood donors recruited for this study. Day 14: O RhD positive platelets (marked with an asterisk) were obtained from Finnish Red Cross Blood Service surplus stock at unit expiry of 5 days or within 12 h afterwards. To create 1:1:1 RWB, 1 RBC unit, 1 Octaplas and ¼ of pooled platelet unit from four donors were mixed. OctaplasLG and lyophilized plasma (LyoPlas N-w) were commercially available blood group AB plasma products. CSWB, cold-stored whole blood; EV, extracellular vesicle; LP, lyophilized plasma; NTA, nanoparticle tracking analysis; PLT, buffy coat platelets; RBC, packed red blood cells; RWB, reconstituted whole blood; SEC, size-exclusion chromatography; WB, whole blood. Created with BioRender.com (licenced to University of Helsinki).

Article Snippet: Control refers to samples drawn immediately prior to donation to determine baseline for variables without established reference values. (A) Particle concentration of EV samples determined with nanoparticle tracking analysis using NanoSight LM14C (Malvern). (B) CD235a positive particles, (C) CD41 positive particles and (D) CD9 positive particles determined using fluorochrome-labelled antibodies with imaging flow cytometer Amnis ImageStream X Mark II (Luminex, Cytek Biosciences).

Techniques: Sampling, Sample Prep, Clinical Proteomics, Size-exclusion Chromatography

Journal: Transfusion medicine (Oxford, England)

Article Title: Extracellular vesicles in ageing cold-stored whole blood may not compensate for the decreasing haemostatic function in vitro.

doi: 10.1111/tme.13122

Figure Lengend Snippet: FIGURE 2 Characterisation of extracellular vesicles. Individual measurements with median are shown, n = 6 for control samples, n = 4 for other groups. Control refers to samples drawn immediately prior to donation to determine baseline for variables without established reference values. (A) Particle concentration of EV samples determined with nanoparticle tracking analysis using NanoSight LM14C (Malvern). (B) CD235a positive particles, (C) CD41 positive particles and (D) CD9 positive particles determined using fluorochrome-labelled antibodies with imaging flow cytometer Amnis ImageStream X Mark II (Luminex, Cytek Biosciences). Changes within each group (i.e. d1 vs. d14) were not significant. Differences between the groups were tested with median test for independent samples with Bonferroni correction for multiple comparisons. CSWB, cold-stored whole blood; RBC+LP packed red blood cells and lyophilized plasma; RWB, reconstituted whole blood.

Article Snippet: Control refers to samples drawn immediately prior to donation to determine baseline for variables without established reference values. (A) Particle concentration of EV samples determined with nanoparticle tracking analysis using NanoSight LM14C (Malvern). (B) CD235a positive particles, (C) CD41 positive particles and (D) CD9 positive particles determined using fluorochrome-labelled antibodies with imaging flow cytometer Amnis ImageStream X Mark II (Luminex, Cytek Biosciences).

Techniques: Control, Concentration Assay, Imaging, Flow Cytometry, Luminex, Clinical Proteomics

Journal: Transfusion medicine (Oxford, England)

Article Title: Extracellular vesicles in ageing cold-stored whole blood may not compensate for the decreasing haemostatic function in vitro.

doi: 10.1111/tme.13122

Figure Lengend Snippet: FIGURE 3 Additional characterisation of RBC+LP and LP samples. Individual measurements with median are shown, n = 4 for RBC+LP and n = 2 for LP. (A) Particle concentration determined with nanoparticle tracking analysis ZetaView PMX-120 (ParticleMetrix), normalised to LP volume. (B) CD235a positive particles determined using fluorochrome-labelled antibodies with imaging flow cytometer Amnis ImageStream X Mark II (Luminex, Cytek Biosciences), normalised to LP volume. (C) SDS-PAGE gel shows the similarity of protein contents in the samples. (D) Western blot showing the absence of CD235a in both RBC+LP and LP samples, positive control included. LP, lyophilized plasma; RBC+LP, packed red blood cells and lyophilized plasma.

Article Snippet: Control refers to samples drawn immediately prior to donation to determine baseline for variables without established reference values. (A) Particle concentration of EV samples determined with nanoparticle tracking analysis using NanoSight LM14C (Malvern). (B) CD235a positive particles, (C) CD41 positive particles and (D) CD9 positive particles determined using fluorochrome-labelled antibodies with imaging flow cytometer Amnis ImageStream X Mark II (Luminex, Cytek Biosciences).

Techniques: Concentration Assay, Imaging, Flow Cytometry, Luminex, SDS Page, Western Blot, Positive Control, Clinical Proteomics