Cargando…

Label‐free imaging of non‐deparaffinized sections of the human kidney to determine tissue quality and signatures of disease

Label‐free fluorescence imaging of kidney sections can provide important morphological information, but its utility has not been tested in a histology processing workflow. We tested the feasibility of label‐free imaging of paraffin‐embedded sections without deparaffinization and its potential useful...

Descripción completa

Detalles Bibliográficos
Autores principales: Sabo, Angela R., Winfree, Seth, Bledsoe, Sharon B., Phillips, Carrie L., Lingeman, James E., Eadon, Michael T., Williams, James C., El‐Achkar, Tarek M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8822874/
https://www.ncbi.nlm.nih.gov/pubmed/35133089
http://dx.doi.org/10.14814/phy2.15167
_version_ 1784646693021548544
author Sabo, Angela R.
Winfree, Seth
Bledsoe, Sharon B.
Phillips, Carrie L.
Lingeman, James E.
Eadon, Michael T.
Williams, James C.
El‐Achkar, Tarek M.
author_facet Sabo, Angela R.
Winfree, Seth
Bledsoe, Sharon B.
Phillips, Carrie L.
Lingeman, James E.
Eadon, Michael T.
Williams, James C.
El‐Achkar, Tarek M.
author_sort Sabo, Angela R.
collection PubMed
description Label‐free fluorescence imaging of kidney sections can provide important morphological information, but its utility has not been tested in a histology processing workflow. We tested the feasibility of label‐free imaging of paraffin‐embedded sections without deparaffinization and its potential usefulness in generating actionable data. Kidney tissue specimens were obtained during percutaneous nephrolithotomy or via diagnostic needle biopsy. Unstained non‐deparaffinized sections were imaged using widefield fluorescence microscopy to capture endogenous fluorescence. Some samples were also imaged with confocal microscopy and multiphoton excitation to collect second harmonic generation (SHG) signal to obtain high‐quality autofluorescence images with optical sectioning. To adjudicate the label‐free signal, the samples or corresponding contiguous sections were subsequently deparaffinized and stained with Lillie's allochrome. Label‐free imaging allowed the recognition of various kidney structures and enabled morphological qualification for adequacy. SHG and confocal imaging yielded quantifiable high‐quality images for tissue collagens and revealed specific patterns in glomeruli and various tubules. Disease specimens from patients with diabetic kidney disease and focal segmental glomerulosclerosis showed distinctive signatures compared to specimens from healthy controls with normal kidney function. Quantitative cytometry could also be performed when DAPI is added in situ before imaging. These results show that label‐free imaging of non‐deparaffinized sections provides useful information about tissue quality that could be beneficial to nephropathologists by maximizing the use of scarce kidney tissue. This approach also provides quantifiable features that could inform on the biology of health and disease.
format Online
Article
Text
id pubmed-8822874
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher John Wiley and Sons Inc.
record_format MEDLINE/PubMed
spelling pubmed-88228742022-02-11 Label‐free imaging of non‐deparaffinized sections of the human kidney to determine tissue quality and signatures of disease Sabo, Angela R. Winfree, Seth Bledsoe, Sharon B. Phillips, Carrie L. Lingeman, James E. Eadon, Michael T. Williams, James C. El‐Achkar, Tarek M. Physiol Rep Original Articles Label‐free fluorescence imaging of kidney sections can provide important morphological information, but its utility has not been tested in a histology processing workflow. We tested the feasibility of label‐free imaging of paraffin‐embedded sections without deparaffinization and its potential usefulness in generating actionable data. Kidney tissue specimens were obtained during percutaneous nephrolithotomy or via diagnostic needle biopsy. Unstained non‐deparaffinized sections were imaged using widefield fluorescence microscopy to capture endogenous fluorescence. Some samples were also imaged with confocal microscopy and multiphoton excitation to collect second harmonic generation (SHG) signal to obtain high‐quality autofluorescence images with optical sectioning. To adjudicate the label‐free signal, the samples or corresponding contiguous sections were subsequently deparaffinized and stained with Lillie's allochrome. Label‐free imaging allowed the recognition of various kidney structures and enabled morphological qualification for adequacy. SHG and confocal imaging yielded quantifiable high‐quality images for tissue collagens and revealed specific patterns in glomeruli and various tubules. Disease specimens from patients with diabetic kidney disease and focal segmental glomerulosclerosis showed distinctive signatures compared to specimens from healthy controls with normal kidney function. Quantitative cytometry could also be performed when DAPI is added in situ before imaging. These results show that label‐free imaging of non‐deparaffinized sections provides useful information about tissue quality that could be beneficial to nephropathologists by maximizing the use of scarce kidney tissue. This approach also provides quantifiable features that could inform on the biology of health and disease. John Wiley and Sons Inc. 2022-02-08 /pmc/articles/PMC8822874/ /pubmed/35133089 http://dx.doi.org/10.14814/phy2.15167 Text en © 2022 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Sabo, Angela R.
Winfree, Seth
Bledsoe, Sharon B.
Phillips, Carrie L.
Lingeman, James E.
Eadon, Michael T.
Williams, James C.
El‐Achkar, Tarek M.
Label‐free imaging of non‐deparaffinized sections of the human kidney to determine tissue quality and signatures of disease
title Label‐free imaging of non‐deparaffinized sections of the human kidney to determine tissue quality and signatures of disease
title_full Label‐free imaging of non‐deparaffinized sections of the human kidney to determine tissue quality and signatures of disease
title_fullStr Label‐free imaging of non‐deparaffinized sections of the human kidney to determine tissue quality and signatures of disease
title_full_unstemmed Label‐free imaging of non‐deparaffinized sections of the human kidney to determine tissue quality and signatures of disease
title_short Label‐free imaging of non‐deparaffinized sections of the human kidney to determine tissue quality and signatures of disease
title_sort label‐free imaging of non‐deparaffinized sections of the human kidney to determine tissue quality and signatures of disease
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8822874/
https://www.ncbi.nlm.nih.gov/pubmed/35133089
http://dx.doi.org/10.14814/phy2.15167
work_keys_str_mv AT saboangelar labelfreeimagingofnondeparaffinizedsectionsofthehumankidneytodeterminetissuequalityandsignaturesofdisease
AT winfreeseth labelfreeimagingofnondeparaffinizedsectionsofthehumankidneytodeterminetissuequalityandsignaturesofdisease
AT bledsoesharonb labelfreeimagingofnondeparaffinizedsectionsofthehumankidneytodeterminetissuequalityandsignaturesofdisease
AT phillipscarriel labelfreeimagingofnondeparaffinizedsectionsofthehumankidneytodeterminetissuequalityandsignaturesofdisease
AT lingemanjamese labelfreeimagingofnondeparaffinizedsectionsofthehumankidneytodeterminetissuequalityandsignaturesofdisease
AT eadonmichaelt labelfreeimagingofnondeparaffinizedsectionsofthehumankidneytodeterminetissuequalityandsignaturesofdisease
AT williamsjamesc labelfreeimagingofnondeparaffinizedsectionsofthehumankidneytodeterminetissuequalityandsignaturesofdisease
AT elachkartarekm labelfreeimagingofnondeparaffinizedsectionsofthehumankidneytodeterminetissuequalityandsignaturesofdisease