Analysis of Neat Biofluids Obtained During Cardiac Surgery Using Nanoparticle Tracking Analysis: Methodological Considerations
Small extracellular vesicles (sEVs) are those nanovesicles 30–150 nm in size with a role in cell signalling and potential as biomarkers of disease. Nanoparticle tracking analysis (NTA) techniques are commonly used to measure sEV concentration in biofluids. However, this quantification technique can...
Main Authors: | , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Frontiers Media S.A.
2020-05-01
|
Series: | Frontiers in Cell and Developmental Biology |
Subjects: | |
Online Access: | https://www.frontiersin.org/article/10.3389/fcell.2020.00367/full |
id |
doaj-6cd858cf2f7d4074b1a303dc745e9b71 |
---|---|
record_format |
Article |
spelling |
doaj-6cd858cf2f7d4074b1a303dc745e9b712020-11-25T03:17:19ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2020-05-01810.3389/fcell.2020.00367495117Analysis of Neat Biofluids Obtained During Cardiac Surgery Using Nanoparticle Tracking Analysis: Methodological ConsiderationsAndrew I. U. Shearn0Sezin Aday1Soumaya Ben-Aicha2Pauline Carnell-Morris3Agnieszka Siupa4Gianni D. Angelini5Aled Clayton6Chantal Boulanger7Prakash Punjabi8Costanza Emanueli9Costanza Emanueli10Giovanni Biglino11Giovanni Biglino12Bristol Heart Institute, Bristol Royal Infirmary, University of Bristol, Bristol, United KingdomBristol Heart Institute, Bristol Royal Infirmary, University of Bristol, Bristol, United KingdomNational Heart and Lung Institute, Imperial College London, London, United KingdomMalvern Instruments Ltd., Malvern, United KingdomMalvern Instruments Ltd., Malvern, United KingdomBristol Heart Institute, Bristol Royal Infirmary, University of Bristol, Bristol, United KingdomSchool of Medicine, Cardiff University, Cardiff, United KingdomCardiovascular Research Center, INSERM U970, Hôpital Européen Georges Pompidou, Paris, FranceNational Heart and Lung Institute, Imperial College London, London, United KingdomBristol Heart Institute, Bristol Royal Infirmary, University of Bristol, Bristol, United KingdomNational Heart and Lung Institute, Imperial College London, London, United KingdomBristol Heart Institute, Bristol Royal Infirmary, University of Bristol, Bristol, United KingdomNational Heart and Lung Institute, Imperial College London, London, United KingdomSmall extracellular vesicles (sEVs) are those nanovesicles 30–150 nm in size with a role in cell signalling and potential as biomarkers of disease. Nanoparticle tracking analysis (NTA) techniques are commonly used to measure sEV concentration in biofluids. However, this quantification technique can be susceptible to sample handing and machine settings. Moreover, some classes of lipoproteins are of similar sizes and could therefore confound sEV quantification, particularly in blood-derived preparations, such serum and plasma. Here we have provided methodological information on NTA measurements and systematically investigated potential factors that could interfere with the reliability and repeatability of results obtained when looking at neat biofluids (i.e., human serum and pericardial fluid) obtained from patients undergoing cardiac surgery and from healthy controls. Data suggest that variables that can affect vesicle quantification include the level of contamination from lipoproteins, number of sample freeze/thaw cycles, sample filtration, using saline-based diluents, video length and keeping the number of particles per frame within defined limits. Those parameters that are of less concern include focus, the “Maximum Jump” setting and the number of videos recorded. However, if these settings are clearly inappropriate the results obtained will be spurious. Similarly, good experimental practice suggests that multiple videos should be recorded. In conclusion, NTA is a perfectible, but still commonly used system for sEVs analyses. Provided users handle their samples with a highly robust and consistent protocol, and accurately report these aspects, they can obtain data that could potentially translate into new clinical biomarkers for diagnosis and monitoring of cardiovascular disease.https://www.frontiersin.org/article/10.3389/fcell.2020.00367/fullcardiac surgeryexosome (vesicle)exosomal biomarkersnanopartciclesNTA (nanoparticle tracking analysis) |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Andrew I. U. Shearn Sezin Aday Soumaya Ben-Aicha Pauline Carnell-Morris Agnieszka Siupa Gianni D. Angelini Aled Clayton Chantal Boulanger Prakash Punjabi Costanza Emanueli Costanza Emanueli Giovanni Biglino Giovanni Biglino |
spellingShingle |
Andrew I. U. Shearn Sezin Aday Soumaya Ben-Aicha Pauline Carnell-Morris Agnieszka Siupa Gianni D. Angelini Aled Clayton Chantal Boulanger Prakash Punjabi Costanza Emanueli Costanza Emanueli Giovanni Biglino Giovanni Biglino Analysis of Neat Biofluids Obtained During Cardiac Surgery Using Nanoparticle Tracking Analysis: Methodological Considerations Frontiers in Cell and Developmental Biology cardiac surgery exosome (vesicle) exosomal biomarkers nanopartcicles NTA (nanoparticle tracking analysis) |
author_facet |
Andrew I. U. Shearn Sezin Aday Soumaya Ben-Aicha Pauline Carnell-Morris Agnieszka Siupa Gianni D. Angelini Aled Clayton Chantal Boulanger Prakash Punjabi Costanza Emanueli Costanza Emanueli Giovanni Biglino Giovanni Biglino |
author_sort |
Andrew I. U. Shearn |
title |
Analysis of Neat Biofluids Obtained During Cardiac Surgery Using Nanoparticle Tracking Analysis: Methodological Considerations |
title_short |
Analysis of Neat Biofluids Obtained During Cardiac Surgery Using Nanoparticle Tracking Analysis: Methodological Considerations |
title_full |
Analysis of Neat Biofluids Obtained During Cardiac Surgery Using Nanoparticle Tracking Analysis: Methodological Considerations |
title_fullStr |
Analysis of Neat Biofluids Obtained During Cardiac Surgery Using Nanoparticle Tracking Analysis: Methodological Considerations |
title_full_unstemmed |
Analysis of Neat Biofluids Obtained During Cardiac Surgery Using Nanoparticle Tracking Analysis: Methodological Considerations |
title_sort |
analysis of neat biofluids obtained during cardiac surgery using nanoparticle tracking analysis: methodological considerations |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Cell and Developmental Biology |
issn |
2296-634X |
publishDate |
2020-05-01 |
description |
Small extracellular vesicles (sEVs) are those nanovesicles 30–150 nm in size with a role in cell signalling and potential as biomarkers of disease. Nanoparticle tracking analysis (NTA) techniques are commonly used to measure sEV concentration in biofluids. However, this quantification technique can be susceptible to sample handing and machine settings. Moreover, some classes of lipoproteins are of similar sizes and could therefore confound sEV quantification, particularly in blood-derived preparations, such serum and plasma. Here we have provided methodological information on NTA measurements and systematically investigated potential factors that could interfere with the reliability and repeatability of results obtained when looking at neat biofluids (i.e., human serum and pericardial fluid) obtained from patients undergoing cardiac surgery and from healthy controls. Data suggest that variables that can affect vesicle quantification include the level of contamination from lipoproteins, number of sample freeze/thaw cycles, sample filtration, using saline-based diluents, video length and keeping the number of particles per frame within defined limits. Those parameters that are of less concern include focus, the “Maximum Jump” setting and the number of videos recorded. However, if these settings are clearly inappropriate the results obtained will be spurious. Similarly, good experimental practice suggests that multiple videos should be recorded. In conclusion, NTA is a perfectible, but still commonly used system for sEVs analyses. Provided users handle their samples with a highly robust and consistent protocol, and accurately report these aspects, they can obtain data that could potentially translate into new clinical biomarkers for diagnosis and monitoring of cardiovascular disease. |
topic |
cardiac surgery exosome (vesicle) exosomal biomarkers nanopartcicles NTA (nanoparticle tracking analysis) |
url |
https://www.frontiersin.org/article/10.3389/fcell.2020.00367/full |
work_keys_str_mv |
AT andrewiushearn analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT sezinaday analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT soumayabenaicha analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT paulinecarnellmorris analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT agnieszkasiupa analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT giannidangelini analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT aledclayton analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT chantalboulanger analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT prakashpunjabi analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT costanzaemanueli analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT costanzaemanueli analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT giovannibiglino analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations AT giovannibiglino analysisofneatbiofluidsobtainedduringcardiacsurgeryusingnanoparticletrackinganalysismethodologicalconsiderations |
_version_ |
1724632047884959744 |