The effects of incubation media on the assessment of the shape of human erythrocytes by flow cytometry: a contribution to mathematical data interpretation to enable wider application of the method
Само за регистроване кориснике
2021
Аутори
Drvenica, IvanaMojsilović, Slavko
Stančić, Ana
Marković, Dragana
Kovačić, Marijana
Maslovarić, Irina
Rapajić, Ivana
Vučetić, Dušan
Ilić, Vesna
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Flow cytometry (FC) analysis of erythrocyte shape and related biomechanical properties, such as osmotic fragility, have not moved from a research tool to regular clinical testing. The main reason is existing evidence that various pre-analytical factors influence the mathematical interpretation of the data obtained. With an aim to contribute to the standardization and broaden the use of FC for human erythrocyte shape assessment, freshly prepared peripheral blood erythrocytes isolated from healthy donors were incubated in iso and hypo-osmotic solutions (pure saline, saline with potassium and calcium, and phosphate buffered saline) and examined by FC using values of forward scatter (FSC) and side scatter (SSC). Kurtosis, skewness, Pearson's second skewness coefficient of dissymmetry (PCD), and spherical index, calculated from FSC distributions, were used for the erythrocyte shape evaluation. In all isotonic media FSC distribution and FSC-based morphology parameters showed huge inter-indiv...idual and inter-medium variation. With decreasing osmolality, in all media and samples, the size of the erythrocytes increased, and swelling index and kurtosis decreased. However, changes in skewness and PCD were influenced by the medium used and the sample tested. Compared to FSC, SSC signal in isotonic and its change in hypotonic media showed lower inter-individual variation and was not influenced by the type of medium. We propose a spherical index and kurtosis as FSC-based indicators of erythrocyte shape. As more resistant to the influence of the preanalytical treatment, SSC data appeared to be unfairly neglected for the assessment of erythrocyte shape, in comparison to the usually employed FSC data.
Кључне речи:
Human erythrocytes / Flow cytometry / Osmotic swelling / Morphology / Sphericity / Multimodal distributionИзвор:
European Biophysics Journal, 2021, 50, 6, 829-846Издавач:
- Springer Nature
Финансирање / пројекти:
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200015 (Универзитет у Београду, Институт за медицинска истраживања) (RS-MESTD-inst-2020-200015)
- HEMMAGINERO - Hemoglobin-Based Spectroscopy and Nonlinear Imaging of Erythrocytes and Their Membranes As Emerging Diagnostic Tool (RS-ScienceFundRS-Promis-6066079)
Институција/група
Institut za medicinska istraživanjaTY - JOUR AU - Drvenica, Ivana AU - Mojsilović, Slavko AU - Stančić, Ana AU - Marković, Dragana AU - Kovačić, Marijana AU - Maslovarić, Irina AU - Rapajić, Ivana AU - Vučetić, Dušan AU - Ilić, Vesna PY - 2021 UR - http://rimi.imi.bg.ac.rs/handle/123456789/1176 AB - Flow cytometry (FC) analysis of erythrocyte shape and related biomechanical properties, such as osmotic fragility, have not moved from a research tool to regular clinical testing. The main reason is existing evidence that various pre-analytical factors influence the mathematical interpretation of the data obtained. With an aim to contribute to the standardization and broaden the use of FC for human erythrocyte shape assessment, freshly prepared peripheral blood erythrocytes isolated from healthy donors were incubated in iso and hypo-osmotic solutions (pure saline, saline with potassium and calcium, and phosphate buffered saline) and examined by FC using values of forward scatter (FSC) and side scatter (SSC). Kurtosis, skewness, Pearson's second skewness coefficient of dissymmetry (PCD), and spherical index, calculated from FSC distributions, were used for the erythrocyte shape evaluation. In all isotonic media FSC distribution and FSC-based morphology parameters showed huge inter-individual and inter-medium variation. With decreasing osmolality, in all media and samples, the size of the erythrocytes increased, and swelling index and kurtosis decreased. However, changes in skewness and PCD were influenced by the medium used and the sample tested. Compared to FSC, SSC signal in isotonic and its change in hypotonic media showed lower inter-individual variation and was not influenced by the type of medium. We propose a spherical index and kurtosis as FSC-based indicators of erythrocyte shape. As more resistant to the influence of the preanalytical treatment, SSC data appeared to be unfairly neglected for the assessment of erythrocyte shape, in comparison to the usually employed FSC data. PB - Springer Nature T2 - European Biophysics Journal T1 - The effects of incubation media on the assessment of the shape of human erythrocytes by flow cytometry: a contribution to mathematical data interpretation to enable wider application of the method EP - 846 IS - 6 SP - 829 VL - 50 DO - 10.1007/s00249-021-01527-3 ER -
@article{ author = "Drvenica, Ivana and Mojsilović, Slavko and Stančić, Ana and Marković, Dragana and Kovačić, Marijana and Maslovarić, Irina and Rapajić, Ivana and Vučetić, Dušan and Ilić, Vesna", year = "2021", abstract = "Flow cytometry (FC) analysis of erythrocyte shape and related biomechanical properties, such as osmotic fragility, have not moved from a research tool to regular clinical testing. The main reason is existing evidence that various pre-analytical factors influence the mathematical interpretation of the data obtained. With an aim to contribute to the standardization and broaden the use of FC for human erythrocyte shape assessment, freshly prepared peripheral blood erythrocytes isolated from healthy donors were incubated in iso and hypo-osmotic solutions (pure saline, saline with potassium and calcium, and phosphate buffered saline) and examined by FC using values of forward scatter (FSC) and side scatter (SSC). Kurtosis, skewness, Pearson's second skewness coefficient of dissymmetry (PCD), and spherical index, calculated from FSC distributions, were used for the erythrocyte shape evaluation. In all isotonic media FSC distribution and FSC-based morphology parameters showed huge inter-individual and inter-medium variation. With decreasing osmolality, in all media and samples, the size of the erythrocytes increased, and swelling index and kurtosis decreased. However, changes in skewness and PCD were influenced by the medium used and the sample tested. Compared to FSC, SSC signal in isotonic and its change in hypotonic media showed lower inter-individual variation and was not influenced by the type of medium. We propose a spherical index and kurtosis as FSC-based indicators of erythrocyte shape. As more resistant to the influence of the preanalytical treatment, SSC data appeared to be unfairly neglected for the assessment of erythrocyte shape, in comparison to the usually employed FSC data.", publisher = "Springer Nature", journal = "European Biophysics Journal", title = "The effects of incubation media on the assessment of the shape of human erythrocytes by flow cytometry: a contribution to mathematical data interpretation to enable wider application of the method", pages = "846-829", number = "6", volume = "50", doi = "10.1007/s00249-021-01527-3" }
Drvenica, I., Mojsilović, S., Stančić, A., Marković, D., Kovačić, M., Maslovarić, I., Rapajić, I., Vučetić, D.,& Ilić, V.. (2021). The effects of incubation media on the assessment of the shape of human erythrocytes by flow cytometry: a contribution to mathematical data interpretation to enable wider application of the method. in European Biophysics Journal Springer Nature., 50(6), 829-846. https://doi.org/10.1007/s00249-021-01527-3
Drvenica I, Mojsilović S, Stančić A, Marković D, Kovačić M, Maslovarić I, Rapajić I, Vučetić D, Ilić V. The effects of incubation media on the assessment of the shape of human erythrocytes by flow cytometry: a contribution to mathematical data interpretation to enable wider application of the method. in European Biophysics Journal. 2021;50(6):829-846. doi:10.1007/s00249-021-01527-3 .
Drvenica, Ivana, Mojsilović, Slavko, Stančić, Ana, Marković, Dragana, Kovačić, Marijana, Maslovarić, Irina, Rapajić, Ivana, Vučetić, Dušan, Ilić, Vesna, "The effects of incubation media on the assessment of the shape of human erythrocytes by flow cytometry: a contribution to mathematical data interpretation to enable wider application of the method" in European Biophysics Journal, 50, no. 6 (2021):829-846, https://doi.org/10.1007/s00249-021-01527-3 . .