Estimation of 100 nm particle distribution kinetics in mouse lung using confocal laser scanning microscopy

Cover Page


Cite item

Full Text

Open Access Open Access
Restricted Access Access granted
Restricted Access Subscription or Fee Access

Abstract

BACKGROUND: Daily, people inhale airborne viral particles, some of which have a size of about 100 nm, such as particles of SARS-CoV-2. Kinetics of such 100 nm particle distribution in the respiratory tract is important, however, not a properly investigated question.

AIM: To estimate the dissemination of inert viral particles based on the analysis of the spatial distribution of fluorescent 100 nm particles in the mouse lungs at different time points after the application.

MATHERIALS AND METHODS: Fluorescent particles of 100 nm size were applied to C57BL/6 mice. 6, 24, 48 and 72 hours after, lungs were excised and fixed. Lung lobes were stained with immunohistochemistry as whole-mounts and then underwent optical clearance. Three-dimensional images of whole-mount mouse lung lobes were acquired using confocal laser scanning microscopy.

RESULTS: 6 hours after the particle application particles were detected in lungs both as single particles and as particle agglomerates. Particles were both free and internalized by phagocytic cells. 24 hours after the application particles were detected both in bronchial lumen and in the alveolar space. Particles were detected in the mouse lungs up to 72 hours after the application.

CONCLUSIONS: Reaching the respiratory tract of mammalian, inert particles which size equal to SARS-CoV-2 particle size distribute both in bronchi and in alveoli and undergoes internalization of phagocytic cells.

Full Text

Restricted Access

About the authors

Julia D. Vavilova

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences

Email: Juliateterina12@gmail.com
ORCID iD: 0000-0002-9075-218X

Postgraduate student, junior scientific researcher

Russian Federation, Moscow

Elena L. Bolkhovitina

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences

Email: alenkash83@gmail.com
ORCID iD: 0000-0002-3386-509X

junior scientific researcher

Russian Federation, Moscow

Andrey O. Bogorodskiy

Moscow Institute of Physics and Technology (National Research University)

Email: bogorodskiy173@gmail.com
ORCID iD: 0000-0002-7589-7823

junior scientific researcher

Russian Federation, Dolgoprudny

Ivan S. Okhrimenko

Moscow Institute of Physics and Technology (National Research University)

Email: ivan.okhrimenko@phystech.edu
ORCID iD: 0000-0002-1053-2778
SPIN-code: 8418-0194

scientific researcher

Russian Federation, Dolgoprudny

Valentin I. Borshchevskiy

Moscow Institute of Physics and Technology (National Research University)

Email: borshchevskiy.vi@phystech.edu
ORCID iD: 0000-0003-4398-9712
SPIN-code: 2018-8957

senior scientific researcher

Russian Federation, Dolgoprudny

Marina A. Shevchenko

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences

Author for correspondence.
Email: mshevch@gmail.com
ORCID iD: 0000-0001-5278-9937
SPIN-code: 8509-1096

scientific researcher

Russian Federation, Moscow

References

  1. Rao G.V., Tinkle S., Weissman D.N. et al. Efficacy of a technique for exposing the mouse lung to particles aspirated from the pharynx // J. Toxicol. Environ. Health A. 2003. Vol. 66, No. 15. P. 1441–1452. doi: 10.1080/15287390306417
  2. Scott G.D., Blum E.D., Fryer A.D., Jacoby D.B. Tissue optical clearing, three-dimensional imaging, and computer morphometry in whole mouse lungs and human airways // Am. J. Respir. Cell. Mol. Biol. 2014. Vol. 1, No. 51. P. 43–55. doi: 10.1165/rcmb.2013-0284OC
  3. Li W., Germain R.N., Gerner M.Y. High-dimensional cell-level analysis of tissues with Ce3D multiplex volume imaging // Nat. Protoc. 2019. Vol. 14, No. 6. P. 1708–1733. doi: 10.1038/s41596-019-0156-4
  4. References
  5. Rao GV, Tinkle S, Weissman DN, et al. Efficacy of a technique for exposing the mouse lung to particles aspirated from the pharynx. J Toxicol Environ Health A. 2003;66(15):1441–1452. doi: 10.1080/15287390306417
  6. Scott GD, Blum ED, Fryer AD, Jacoby DB. Tissue optical clearing, three-dimensional imaging, and computer morphometry in whole mouse lungs and human airways. Am J Respir Cell Mol Biol. 2014;1(51):43–55. doi: 10.1165/rcmb.2013-0284OC
  7. Li W, Germain RN, Gerner MY. High-dimensional cell-level analysis of tissues with Ce3D multiplex volume imaging. Nat Protoc. 2019;14(6):1708–1733. doi: 10.1038/s41596-019-0156-4

Supplementary files

Supplementary Files
Action
1. Fig. 1. Distribution of 100 nm fluorescent particles in the airways: a — the representative image of the whole-mount mouse lung 24 hours after oropharyngeal application of fluorescent particles (magenta). Airways are stained with streptavidin (greyscale). Scale bar 1000 µm; b — enlarged image of the region indicated with a dotted square in a. Particles in the bronchial branches are indicated with arrows, in the alveolar space with arrowheads. Scale bar 100 µm

Download (392KB)
2. Fig. 2. Different states of 100 nm particles in the airways: a, b — representative images of particles (magenta) in the whole-mount mouse lung 24 hours after the application. Scale bar 40 µm; c, d — enlarged image of the region indicated in a. Arrows indicate particles internalized by cells; arrowheads free particles Scale bar 20 µm. Actin filaments are stained with streptavidin (greyscale) (a, c, d)

Download (406KB)
3. Fig. 3. 100 nm particles agglomerate formation in mouse lungs. Representative images of the regions of whole-mount lung specimens at 6 (a), 24 (b), 48 (c) and 72 hours (d) after particle application. Airway wall (streptavidin, greyscale), particles (magenta). Scale bar 100 µm

Download (840KB)

Copyright (c) 2021 Vavilova J.D., Bolkhovitina E.L., Bogorodskiy A.O., Okhrimenko I.S., Borshchevskiy V.I., Shevchenko M.A.

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: серия ПИ № ФС 77 - 74760 от 29.12.2018 г.


This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies