Assessment of small cellular particles from four different natural sources and liposomes by interferometric light microscopy

Romolo, Anna (author), Jan, Zala (author), Bedina Zavec, Apolonija (author), Kisovec, Matic (author), Arrigler, Vesna (author), Spasovski, Vesna (author), Podobnik, Marjetka (author), Iglič, Aleš (author), Pocsfalvi, Gabriella (author), Kogej, Ksenija (author), Kralj-Iglič, Veronika (author)

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.

• Language: English
• Keywords: interferometric light microscopy, interferometric nanotracking analysis, nanoparticle tracking analysis, concentration of extracellular particles, size of extracellular particles, dynamic light scattering, hydrodynamic radius of extracellular particles, extracellular vesicles, liposomes, exosomes
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: ZF - Faculty of Health Sciences; FE - Faculty of Electrical Engineering; FKKT - Faculty of Chemistry and Chemical Technology; MF - Faculty of Medicine
• Publication status: Published
• Publication version: Version of Record
• Publication date: 13.12.2022
• Year: 2022
• Number of pages: 22 str.
• Numbering: Vol. 23, no. 24, art. 15801
• PID: 20.500.12556/RUL-143699
• UDC: 577:543.456
• ISSN on article: 1422-0067
• DOI: 10.3390/ijms232415801
• COBISS.SI-ID: 136801795
• Publication date in RUL: 09.01.2023

Record is a part of a journal

• Title: International journal of molecular sciences
• Shortened title: Int. j. mol. sci.
• Publisher: MDPI
• ISSN: 1422-0067
• COBISS.SI-ID: 2779162

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
• Licensing start date: 13.12.2022

Secondary language

• Language: Undetermined
• Keywords: interferometrična svetlobna mikroskopija, interferometrična nanosledilna analiza, analiza sledenja nanodelcev, koncentracija zunajceličnih delcev, velikost zunajceličnih delcev, dinamično sipanje svetlobe, hidrodinamični radij zunajceličnih delcev, zunajcelični vezikli, liposomi, eksosomi