High resolution fluorescence imaging of the alveolar scaffold as a novel tool to assess lung injury
Lindstedt, Sandra LU ; Wang, Qi LU ; Niroomand, Anna LU ; Stenlo, Martin LU ; Hyllen, Snejana LU ; Pierre, Leif LU ; Olm, Franziska LU
and Bechet, Nicholas B
LU
(2024)
In Scientific Reports
14(1).
- Abstract
Acute lung injury (ALI) represents an aetiologically diverse form of pulmonary damage. Part of the assessment and diagnosis of ALI depends on skilled observer-based scoring of brightfield microscopy tissue sections. Although this readout is sufficient to determine gross alterations in tissue structure, its categorical scores lack the sensitivity to describe more subtle changes in lung morphology. To generate a more sensitive readout of alveolar perturbation we carried out high resolution immunofluorescence imaging on 200 μm lung vibratome sections from baseline and acutely injured porcine lung tissue, stained with a tomato lectin, Lycopersicon Esculentum Dylight-488. With the ability to resolve individual alveoli along with their inner... (More)
Acute lung injury (ALI) represents an aetiologically diverse form of pulmonary damage. Part of the assessment and diagnosis of ALI depends on skilled observer-based scoring of brightfield microscopy tissue sections. Although this readout is sufficient to determine gross alterations in tissue structure, its categorical scores lack the sensitivity to describe more subtle changes in lung morphology. To generate a more sensitive readout of alveolar perturbation we carried out high resolution immunofluorescence imaging on 200 μm lung vibratome sections from baseline and acutely injured porcine lung tissue, stained with a tomato lectin, Lycopersicon Esculentum Dylight-488. With the ability to resolve individual alveoli along with their inner and outer wall we generated continuous readouts of alveolar wall thickness and circularity. From 212 alveoli traced from 10 baseline lung samples we established normal distributions for alveolar wall thickness (27.37; 95% CI [26.48:28.26]) and circularity (0.8609; 95% CI [0.8482:0.8667]) in healthy tissue. Compared to acutely injured lung tissue baseline tissue exhibited a significantly lower wall thickness (26.86 ± 0.4998 vs 50.55 ± 4.468; p = 0.0003) and higher degree of circularityϕ≤ (0.8783 ± 0.01965 vs 0.4133 ± 0.04366; p < 0.0001). These two components were subsequently combined into a single more sensitive variable, termed the morphological quotient (MQ), which exhibited a significant negative correlation (R
(Less)
2 = 0.9919, p < 0.0001) with the gold standard of observer-based scoring. Through the utilisation of advanced light imaging we show it is possible to generate sensitive continuous datasets describing fundamental morphological changes that arise in acute lung injury. These data represent valuable new analytical tools that can be used to precisely benchmark changes in alveolar morphology both in disease/injury as well as in response to treatment/therapy.
- author
- Lindstedt, Sandra
LU
; Wang, Qi
LU
; Niroomand, Anna
LU
; Stenlo, Martin
LU
; Hyllen, Snejana
LU
; Pierre, Leif
LU
; Olm, Franziska
LU
and Bechet, Nicholas B
LU
- organization
- publishing date
- 2024
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Biological techniques, Fluorescence imaging, Imaging
- in
- Scientific Reports
- volume
- 14
- issue
- 1
- article number
- 6662
- pages
- 10 pages
- publisher
- Nature Publishing Group
- external identifiers
-
- scopus:85188086870
- pmid:38509285
- ISSN
- 2045-2322
- DOI
- 10.1038/s41598-024-57313-6
- language
- English
- LU publication?
- yes
- id
- 417d0f9f-645f-4e75-8596-0fc6b36b8c1c
- date added to LUP
- 2024-03-22 10:05:02
- date last changed
- 2024-04-18 13:19:27
@article{417d0f9f-645f-4e75-8596-0fc6b36b8c1c,
abstract = {{<p>Acute lung injury (ALI) represents an aetiologically diverse form of pulmonary damage. Part of the assessment and diagnosis of ALI depends on skilled observer-based scoring of brightfield microscopy tissue sections. Although this readout is sufficient to determine gross alterations in tissue structure, its categorical scores lack the sensitivity to describe more subtle changes in lung morphology. To generate a more sensitive readout of alveolar perturbation we carried out high resolution immunofluorescence imaging on 200 μm lung vibratome sections from baseline and acutely injured porcine lung tissue, stained with a tomato lectin, Lycopersicon Esculentum Dylight-488. With the ability to resolve individual alveoli along with their inner and outer wall we generated continuous readouts of alveolar wall thickness and circularity. From 212 alveoli traced from 10 baseline lung samples we established normal distributions for alveolar wall thickness (27.37; 95% CI [26.48:28.26]) and circularity (0.8609; 95% CI [0.8482:0.8667]) in healthy tissue. Compared to acutely injured lung tissue baseline tissue exhibited a significantly lower wall thickness (26.86 ± 0.4998 vs 50.55 ± 4.468; p = 0.0003) and higher degree of circularityϕ≤ (0.8783 ± 0.01965 vs 0.4133 ± 0.04366; p < 0.0001). These two components were subsequently combined into a single more sensitive variable, termed the morphological quotient (MQ), which exhibited a significant negative correlation (R<br>
2 = 0.9919, p < 0.0001) with the gold standard of observer-based scoring. Through the utilisation of advanced light imaging we show it is possible to generate sensitive continuous datasets describing fundamental morphological changes that arise in acute lung injury. These data represent valuable new analytical tools that can be used to precisely benchmark changes in alveolar morphology both in disease/injury as well as in response to treatment/therapy.<br>
</p>}},
author = {{Lindstedt, Sandra and Wang, Qi and Niroomand, Anna and Stenlo, Martin and Hyllen, Snejana and Pierre, Leif and Olm, Franziska and Bechet, Nicholas B}},
issn = {{2045-2322}},
keywords = {{Biological techniques; Fluorescence imaging; Imaging}},
language = {{eng}},
number = {{1}},
publisher = {{Nature Publishing Group}},
series = {{Scientific Reports}},
title = {{High resolution fluorescence imaging of the alveolar scaffold as a novel tool to assess lung injury}},
url = {{http://dx.doi.org/10.1038/s41598-024-57313-6}},
doi = {{10.1038/s41598-024-57313-6}},
volume = {{14}},
year = {{2024}},
}