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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV250007	4/6	Homo sapiens	Blood plasma	(d)(U)C	Korenjak B	2024	17%
Study summary Full title Assessment of Extracellular Particles Directly in Diluted Plasma and Blood by Interferometric Light Microscopy. A Study of 613 Human and 163 Canine Samples. All authors Korenjak B, Tratenšek A, Arko M, Romolo A, Hočevar M, Kisovec M, Berry M, Bedina Zavec A, Drobne D, Vovk T, Iglič A, Nemec Svete A, Erjavec V, Kralj-Iglič V Journal Cells Abstract Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as (show more...)Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as mediators in cell-cell communication/ however, their harvesting and assessment from bodily fluids are challenging, as processing can significantly affect samples. With the aim of minimizing processing artifacts, we assessed the number density () and hydrodynamic diameter () of EPs directly in diluted plasma and blood using the following recently developed technique: interferometric light microscopy (ILM). We analyzed 613 blood and plasma samples from human patients with inflammatory bowel disease (IBD), collected in trisodium citrate and ethylenediaminetetraacetic acid (EDTA) anticoagulants, and 163 blood and plasma samples from canine patients with brachycephalic obstructive airway syndrome (BOAS). We found a highly statistically significant correlation between in the plasma and in the blood only in the human (i.e., but not canine) blood samples, between the samples with trisodium citrate and EDTA, and between the respective for both species (all < 10). In the human plasma, the average <> was 139 ± 31 nm/ in the human blood, <> was 158 ± 11 nm/ in the canine plasma, <> was 155 ± 32 nm/ and in the canine blood, <> was 171 ± 33 nm. The differences within species were statistically significant ( < 10), with sufficient statistical power (P > 0.8). For <>, we found no statistically significant differences between the human plasma and blood samples or between the samples with trisodium citrate and EDTA. Our results prove that measuring and of EPs in minimally processed fresh blood and in diluted fresh plasma by means of ILM is feasible for large populations of samples. (hide) EV-METRIC 17% (45th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Blood plasma Sample origin Inflammatory Bowel Disease Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Function/New methodological development Sample Species Homo sapiens Sample Type Blood plasma Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis EM EM-type Cryo-EM/ Scanning-EM Image type Close-up Other particle analysis name(1) Interferometric light microscopy Report type Median Report size 160-200 EV-concentration Yes Particle yield number of particles per milliliter of starting sample:

	EV250007	1/6	Canis familiaris	Blood plasma	(d)(U)C	Korenjak B	2024	0%
Study summary Full title Assessment of Extracellular Particles Directly in Diluted Plasma and Blood by Interferometric Light Microscopy. A Study of 613 Human and 163 Canine Samples. All authors Korenjak B, Tratenšek A, Arko M, Romolo A, Hočevar M, Kisovec M, Berry M, Bedina Zavec A, Drobne D, Vovk T, Iglič A, Nemec Svete A, Erjavec V, Kralj-Iglič V Journal Cells Abstract Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as (show more...)Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as mediators in cell-cell communication/ however, their harvesting and assessment from bodily fluids are challenging, as processing can significantly affect samples. With the aim of minimizing processing artifacts, we assessed the number density () and hydrodynamic diameter () of EPs directly in diluted plasma and blood using the following recently developed technique: interferometric light microscopy (ILM). We analyzed 613 blood and plasma samples from human patients with inflammatory bowel disease (IBD), collected in trisodium citrate and ethylenediaminetetraacetic acid (EDTA) anticoagulants, and 163 blood and plasma samples from canine patients with brachycephalic obstructive airway syndrome (BOAS). We found a highly statistically significant correlation between in the plasma and in the blood only in the human (i.e., but not canine) blood samples, between the samples with trisodium citrate and EDTA, and between the respective for both species (all < 10). In the human plasma, the average <> was 139 ± 31 nm/ in the human blood, <> was 158 ± 11 nm/ in the canine plasma, <> was 155 ± 32 nm/ and in the canine blood, <> was 171 ± 33 nm. The differences within species were statistically significant ( < 10), with sufficient statistical power (P > 0.8). For <>, we found no statistically significant differences between the human plasma and blood samples or between the samples with trisodium citrate and EDTA. Our results prove that measuring and of EPs in minimally processed fresh blood and in diluted fresh plasma by means of ILM is feasible for large populations of samples. (hide) EV-METRIC 0% (median: 22% of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Blood plasma Sample origin brachycephalic obstructive airway syndrome Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Function/New methodological development Sample Species Canis familiaris Sample Type Blood plasma Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis Other particle analysis name(1) interferometric light microscopy Report type Median Report size 160 EV-concentration Yes Particle yield count of particles/ mL:

	EV250007	2/6	Canis familiaris	Blood plasma	(d)(U)C	Korenjak B	2024	0%
Study summary Full title Assessment of Extracellular Particles Directly in Diluted Plasma and Blood by Interferometric Light Microscopy. A Study of 613 Human and 163 Canine Samples. All authors Korenjak B, Tratenšek A, Arko M, Romolo A, Hočevar M, Kisovec M, Berry M, Bedina Zavec A, Drobne D, Vovk T, Iglič A, Nemec Svete A, Erjavec V, Kralj-Iglič V Journal Cells Abstract Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as (show more...)Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as mediators in cell-cell communication/ however, their harvesting and assessment from bodily fluids are challenging, as processing can significantly affect samples. With the aim of minimizing processing artifacts, we assessed the number density () and hydrodynamic diameter () of EPs directly in diluted plasma and blood using the following recently developed technique: interferometric light microscopy (ILM). We analyzed 613 blood and plasma samples from human patients with inflammatory bowel disease (IBD), collected in trisodium citrate and ethylenediaminetetraacetic acid (EDTA) anticoagulants, and 163 blood and plasma samples from canine patients with brachycephalic obstructive airway syndrome (BOAS). We found a highly statistically significant correlation between in the plasma and in the blood only in the human (i.e., but not canine) blood samples, between the samples with trisodium citrate and EDTA, and between the respective for both species (all < 10). In the human plasma, the average <> was 139 ± 31 nm/ in the human blood, <> was 158 ± 11 nm/ in the canine plasma, <> was 155 ± 32 nm/ and in the canine blood, <> was 171 ± 33 nm. The differences within species were statistically significant ( < 10), with sufficient statistical power (P > 0.8). For <>, we found no statistically significant differences between the human plasma and blood samples or between the samples with trisodium citrate and EDTA. Our results prove that measuring and of EPs in minimally processed fresh blood and in diluted fresh plasma by means of ILM is feasible for large populations of samples. (hide) EV-METRIC 0% (median: 22% of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Blood plasma Sample origin brachycephalic obstructive airway syndrome Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Function/New methodological development Sample Species Canis familiaris Sample Type Blood plasma Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis Other particle analysis name(1) interferometric light microscopy Report type Median Report size 160 EV-concentration Yes Particle yield count of particles/ mL:

	EV250007	3/6	Canis familiaris	Whole blood	(d)(U)C	Korenjak B	2024	0%
Study summary Full title Assessment of Extracellular Particles Directly in Diluted Plasma and Blood by Interferometric Light Microscopy. A Study of 613 Human and 163 Canine Samples. All authors Korenjak B, Tratenšek A, Arko M, Romolo A, Hočevar M, Kisovec M, Berry M, Bedina Zavec A, Drobne D, Vovk T, Iglič A, Nemec Svete A, Erjavec V, Kralj-Iglič V Journal Cells Abstract Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as (show more...)Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as mediators in cell-cell communication/ however, their harvesting and assessment from bodily fluids are challenging, as processing can significantly affect samples. With the aim of minimizing processing artifacts, we assessed the number density () and hydrodynamic diameter () of EPs directly in diluted plasma and blood using the following recently developed technique: interferometric light microscopy (ILM). We analyzed 613 blood and plasma samples from human patients with inflammatory bowel disease (IBD), collected in trisodium citrate and ethylenediaminetetraacetic acid (EDTA) anticoagulants, and 163 blood and plasma samples from canine patients with brachycephalic obstructive airway syndrome (BOAS). We found a highly statistically significant correlation between in the plasma and in the blood only in the human (i.e., but not canine) blood samples, between the samples with trisodium citrate and EDTA, and between the respective for both species (all < 10). In the human plasma, the average <> was 139 ± 31 nm/ in the human blood, <> was 158 ± 11 nm/ in the canine plasma, <> was 155 ± 32 nm/ and in the canine blood, <> was 171 ± 33 nm. The differences within species were statistically significant ( < 10), with sufficient statistical power (P > 0.8). For <>, we found no statistically significant differences between the human plasma and blood samples or between the samples with trisodium citrate and EDTA. Our results prove that measuring and of EPs in minimally processed fresh blood and in diluted fresh plasma by means of ILM is feasible for large populations of samples. (hide) EV-METRIC 0% (median: 0% of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Whole blood Sample origin brachycephalic obstructive airway syndrome Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Function/New methodological development Sample Species Canis familiaris Sample Type Whole blood Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis Other particle analysis name(1) interferometric light microscopy Report type Median Report size 160 EV-concentration Yes Particle yield number of particles per milliliter:

	EV250007	5/6	Homo sapiens	Blood plasma	(d)(U)C	Korenjak B	2024	0%
Study summary Full title Assessment of Extracellular Particles Directly in Diluted Plasma and Blood by Interferometric Light Microscopy. A Study of 613 Human and 163 Canine Samples. All authors Korenjak B, Tratenšek A, Arko M, Romolo A, Hočevar M, Kisovec M, Berry M, Bedina Zavec A, Drobne D, Vovk T, Iglič A, Nemec Svete A, Erjavec V, Kralj-Iglič V Journal Cells Abstract Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as (show more...)Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as mediators in cell-cell communication/ however, their harvesting and assessment from bodily fluids are challenging, as processing can significantly affect samples. With the aim of minimizing processing artifacts, we assessed the number density () and hydrodynamic diameter () of EPs directly in diluted plasma and blood using the following recently developed technique: interferometric light microscopy (ILM). We analyzed 613 blood and plasma samples from human patients with inflammatory bowel disease (IBD), collected in trisodium citrate and ethylenediaminetetraacetic acid (EDTA) anticoagulants, and 163 blood and plasma samples from canine patients with brachycephalic obstructive airway syndrome (BOAS). We found a highly statistically significant correlation between in the plasma and in the blood only in the human (i.e., but not canine) blood samples, between the samples with trisodium citrate and EDTA, and between the respective for both species (all < 10). In the human plasma, the average <> was 139 ± 31 nm/ in the human blood, <> was 158 ± 11 nm/ in the canine plasma, <> was 155 ± 32 nm/ and in the canine blood, <> was 171 ± 33 nm. The differences within species were statistically significant ( < 10), with sufficient statistical power (P > 0.8). For <>, we found no statistically significant differences between the human plasma and blood samples or between the samples with trisodium citrate and EDTA. Our results prove that measuring and of EPs in minimally processed fresh blood and in diluted fresh plasma by means of ILM is feasible for large populations of samples. (hide) EV-METRIC 0% (median: 22% of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Blood plasma Sample origin Inflammatory Bowel Disease Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Function/New methodological development Sample Species Homo sapiens Sample Type Blood plasma Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis Other particle analysis name(1) interferometric light microscopy Report type Median Report size 160 - 200 EV-concentration Yes Particle yield as number of particles per milliliter of starting sample: 5.0 x 10_ - 5.0 x 10_ / milliliter

	EV250007	6/6	Homo sapiens	whole blood	(d)(U)C	Korenjak B	2024	0%
Study summary Full title Assessment of Extracellular Particles Directly in Diluted Plasma and Blood by Interferometric Light Microscopy. A Study of 613 Human and 163 Canine Samples. All authors Korenjak B, Tratenšek A, Arko M, Romolo A, Hočevar M, Kisovec M, Berry M, Bedina Zavec A, Drobne D, Vovk T, Iglič A, Nemec Svete A, Erjavec V, Kralj-Iglič V Journal Cells Abstract Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as (show more...)Extracellular nanoparticles (EPs) are a subject of increasing interest for their biological role as mediators in cell-cell communication/ however, their harvesting and assessment from bodily fluids are challenging, as processing can significantly affect samples. With the aim of minimizing processing artifacts, we assessed the number density () and hydrodynamic diameter () of EPs directly in diluted plasma and blood using the following recently developed technique: interferometric light microscopy (ILM). We analyzed 613 blood and plasma samples from human patients with inflammatory bowel disease (IBD), collected in trisodium citrate and ethylenediaminetetraacetic acid (EDTA) anticoagulants, and 163 blood and plasma samples from canine patients with brachycephalic obstructive airway syndrome (BOAS). We found a highly statistically significant correlation between in the plasma and in the blood only in the human (i.e., but not canine) blood samples, between the samples with trisodium citrate and EDTA, and between the respective for both species (all < 10). In the human plasma, the average <> was 139 ± 31 nm/ in the human blood, <> was 158 ± 11 nm/ in the canine plasma, <> was 155 ± 32 nm/ and in the canine blood, <> was 171 ± 33 nm. The differences within species were statistically significant ( < 10), with sufficient statistical power (P > 0.8). For <>, we found no statistically significant differences between the human plasma and blood samples or between the samples with trisodium citrate and EDTA. Our results prove that measuring and of EPs in minimally processed fresh blood and in diluted fresh plasma by means of ILM is feasible for large populations of samples. (hide) EV-METRIC 0% (median: 0% of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type whole blood Sample origin Inflammatory Bowel Disease Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Function/New methodological development Sample Species Homo sapiens Sample Type whole blood Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Characterization: Protein analysis None Protein Concentration Method Not determined Characterization: Lipid analysis No Characterization: Particle analysis Other particle analysis name(1) interferometric light microscopy Report type Median Report size 160 - 200 EV-concentration Yes Particle yield as number of particles per milliliter of starting sample: 5.0 x 10_ - 5.0 x 10_/ milliliter

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EV-TRACK ID	EV250007
species	Homo sapiens	Canis familiaris	Canis familiaris	Canis familiaris	Homo sapiens	Homo sapiens
sample type	Blood plasma	Blood plasma	Blood plasma	Whole blood	Blood plasma	whole blood
condition	Inflammatory Bowel Disease	brachycephalic obstructive airway syndrome	brachycephalic obstructive airway syndrome	brachycephalic obstructive airway syndrome	Inflammatory Bowel Disease	Inflammatory Bowel Disease
separation protocol	dUC	dUC	dUC	dUC	dUC	dUC
Exp. nr.	4	1	2	3	5	6
EV-METRIC %	17	0	0	0	0	0