Assessment of Small Cellular Particles from Four Different Natural Sources and Liposomes by Interferometric Light Microscopy
Authors
Romolo, AnnaJan, Zala
Bedina Zavec, Apolonija
Kisovec, Matic
Arrigler, Vesna
Spasovski, Vesna
Podobnik, Marjetka
Iglič, Aleš
Pocsfalvi, Gabriella
Kogej, Ksenija
Kralj-Iglič, Veronika
Article (Published version)
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Small particles in natural sources are a subject of interest for their potential role in intercellular, inter-organism, and inter-species interactions, but their harvesting and assessment present a challenge due to their small size and transient identity. We applied a recently developed interferometric light microscopy (ILM) to assess the number density and hydrodynamic radius (Rh) of isolated small cellular particles (SCPs) from blood preparations (plasma and washed erythrocytes) (B), spruce needle homogenate (S), suspension of flagellae of microalgae Tetraselmis chuii (T), conditioned culture media of microalgae Phaeodactylum tricornutum (P), and liposomes (L). The aliquots were also assessed by flow cytometry (FCM), dynamic light scattering (DLS), ultraviolet-visible spectrometry (UV-vis), and imaging by cryogenic transmission electron microscopy (cryo-TEM). In Rh, ILM showed agreement with DLS within the measurement error in 10 out of 13 samples and was the only method used here th...at yielded particle density. Cryo-TEM revealed that representative SCPs from Tetraselmis chuii flagella (T) did not have a globular shape, so the interpretation by Rh of the batch methods was biased. Cryo-TEM showed the presence of thin filaments in isolates from Phaeodactylum tricornutum conditioned culture media (P), which provides an explanation for the considerably larger Rh obtained by batch methods than the sizes of particles observed by cryo-TEM images. ILM proved convenient for assessment of number density and Rh of SCPs in blood preparations (e.g., plasma); therefore, its use in population and clinical studies is indicated.
Keywords:
concentration of extracellular particles / dynamic light scattering / exosomes / extracellular vesicles / hydrodynamic radius of extracellular particles / interferometric light microscopy / interferometric nanotracking analysis / liposomes / nanoparticle tracking analysis / size of extracellular particlesSource:
International Journal of Molecular Sciences, 2022, 23, 24, 15801-Publisher:
- MDPI
Funding / projects:
- Slovenian Research Agency, grant number P2-0232
- Slovenian Research Agency, grant number P3-0388
- Slovenian Research Agency, grant number P1-0201
- Slovenian Research Agency, grant number P1-0391
- Slovenian Research Agency, grant number J3-3066
- Slovenian Research Agency, grant number L3-2621
- Slovenian Research Agency, grant number IO-0003
Institution/Community
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Romolo, Anna AU - Jan, Zala AU - Bedina Zavec, Apolonija AU - Kisovec, Matic AU - Arrigler, Vesna AU - Spasovski, Vesna AU - Podobnik, Marjetka AU - Iglič, Aleš AU - Pocsfalvi, Gabriella AU - Kogej, Ksenija AU - Kralj-Iglič, Veronika PY - 2022 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/2090 AB - Small particles in natural sources are a subject of interest for their potential role in intercellular, inter-organism, and inter-species interactions, but their harvesting and assessment present a challenge due to their small size and transient identity. We applied a recently developed interferometric light microscopy (ILM) to assess the number density and hydrodynamic radius (Rh) of isolated small cellular particles (SCPs) from blood preparations (plasma and washed erythrocytes) (B), spruce needle homogenate (S), suspension of flagellae of microalgae Tetraselmis chuii (T), conditioned culture media of microalgae Phaeodactylum tricornutum (P), and liposomes (L). The aliquots were also assessed by flow cytometry (FCM), dynamic light scattering (DLS), ultraviolet-visible spectrometry (UV-vis), and imaging by cryogenic transmission electron microscopy (cryo-TEM). In Rh, ILM showed agreement with DLS within the measurement error in 10 out of 13 samples and was the only method used here that yielded particle density. Cryo-TEM revealed that representative SCPs from Tetraselmis chuii flagella (T) did not have a globular shape, so the interpretation by Rh of the batch methods was biased. Cryo-TEM showed the presence of thin filaments in isolates from Phaeodactylum tricornutum conditioned culture media (P), which provides an explanation for the considerably larger Rh obtained by batch methods than the sizes of particles observed by cryo-TEM images. ILM proved convenient for assessment of number density and Rh of SCPs in blood preparations (e.g., plasma); therefore, its use in population and clinical studies is indicated. PB - MDPI T2 - International Journal of Molecular Sciences T1 - Assessment of Small Cellular Particles from Four Different Natural Sources and Liposomes by Interferometric Light Microscopy IS - 24 SP - 15801 VL - 23 DO - 10.3390/ijms232415801 ER -
@article{ author = "Romolo, Anna and Jan, Zala and Bedina Zavec, Apolonija and Kisovec, Matic and Arrigler, Vesna and Spasovski, Vesna and Podobnik, Marjetka and Iglič, Aleš and Pocsfalvi, Gabriella and Kogej, Ksenija and Kralj-Iglič, Veronika", year = "2022", abstract = "Small particles in natural sources are a subject of interest for their potential role in intercellular, inter-organism, and inter-species interactions, but their harvesting and assessment present a challenge due to their small size and transient identity. We applied a recently developed interferometric light microscopy (ILM) to assess the number density and hydrodynamic radius (Rh) of isolated small cellular particles (SCPs) from blood preparations (plasma and washed erythrocytes) (B), spruce needle homogenate (S), suspension of flagellae of microalgae Tetraselmis chuii (T), conditioned culture media of microalgae Phaeodactylum tricornutum (P), and liposomes (L). The aliquots were also assessed by flow cytometry (FCM), dynamic light scattering (DLS), ultraviolet-visible spectrometry (UV-vis), and imaging by cryogenic transmission electron microscopy (cryo-TEM). In Rh, ILM showed agreement with DLS within the measurement error in 10 out of 13 samples and was the only method used here that yielded particle density. Cryo-TEM revealed that representative SCPs from Tetraselmis chuii flagella (T) did not have a globular shape, so the interpretation by Rh of the batch methods was biased. Cryo-TEM showed the presence of thin filaments in isolates from Phaeodactylum tricornutum conditioned culture media (P), which provides an explanation for the considerably larger Rh obtained by batch methods than the sizes of particles observed by cryo-TEM images. ILM proved convenient for assessment of number density and Rh of SCPs in blood preparations (e.g., plasma); therefore, its use in population and clinical studies is indicated.", publisher = "MDPI", journal = "International Journal of Molecular Sciences", title = "Assessment of Small Cellular Particles from Four Different Natural Sources and Liposomes by Interferometric Light Microscopy", number = "24", pages = "15801", volume = "23", doi = "10.3390/ijms232415801" }
Romolo, A., Jan, Z., Bedina Zavec, A., Kisovec, M., Arrigler, V., Spasovski, V., Podobnik, M., Iglič, A., Pocsfalvi, G., Kogej, K.,& Kralj-Iglič, V.. (2022). Assessment of Small Cellular Particles from Four Different Natural Sources and Liposomes by Interferometric Light Microscopy. in International Journal of Molecular Sciences MDPI., 23(24), 15801. https://doi.org/10.3390/ijms232415801
Romolo A, Jan Z, Bedina Zavec A, Kisovec M, Arrigler V, Spasovski V, Podobnik M, Iglič A, Pocsfalvi G, Kogej K, Kralj-Iglič V. Assessment of Small Cellular Particles from Four Different Natural Sources and Liposomes by Interferometric Light Microscopy. in International Journal of Molecular Sciences. 2022;23(24):15801. doi:10.3390/ijms232415801 .
Romolo, Anna, Jan, Zala, Bedina Zavec, Apolonija, Kisovec, Matic, Arrigler, Vesna, Spasovski, Vesna, Podobnik, Marjetka, Iglič, Aleš, Pocsfalvi, Gabriella, Kogej, Ksenija, Kralj-Iglič, Veronika, "Assessment of Small Cellular Particles from Four Different Natural Sources and Liposomes by Interferometric Light Microscopy" in International Journal of Molecular Sciences, 23, no. 24 (2022):15801, https://doi.org/10.3390/ijms232415801 . .