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Publication
Label-free imaging of non-deparaffinized sections of the human kidney to
determine tissue quality and signatures of disease.
Authors Sabo AR, Winfree S, Bledsoe SB, Phillips CL, Lingeman JE, Eadon MT, Williams JC,
El-Achkar TM
Submitted By Submitted Externally on 3/9/2022
Status Published
Journal Physiological reports, References
Year 2022
Date Published 2/1/2022
Volume : Pages 10 : e15167
PubMed Reference 35133089
Abstract 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.




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