Label-Free Analysis of Urine Samples with In-Flow Digital Holographic Microscopy

Urinary tract infections are among the most frequent infectious diseases and require screening a great amount of urine samples from patients. However, a high percentage of samples result as negative after urine culture plate tests (CPTs), demanding a simple and fast preliminary technique to screen o...

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Main Authors: Lucia Gigli, Nicoletta Braidotti, Maria Augusta do R. B. F. Lima, Catalin Dacian Ciubotaru, Dan Cojoc
Format: Article
Language:English
Published: MDPI AG 2023-08-01
Series:Biosensors
Subjects:
Online Access:https://www.mdpi.com/2079-6374/13/8/789
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author Lucia Gigli
Nicoletta Braidotti
Maria Augusta do R. B. F. Lima
Catalin Dacian Ciubotaru
Dan Cojoc
author_facet Lucia Gigli
Nicoletta Braidotti
Maria Augusta do R. B. F. Lima
Catalin Dacian Ciubotaru
Dan Cojoc
author_sort Lucia Gigli
collection DOAJ
description Urinary tract infections are among the most frequent infectious diseases and require screening a great amount of urine samples from patients. However, a high percentage of samples result as negative after urine culture plate tests (CPTs), demanding a simple and fast preliminary technique to screen out the negative samples. We propose a digital holographic microscopy (DHM) method to inspect fresh urine samples flowing in a glass capillary for 3 min, recording holograms at 2 frames per second. After digital reconstruction, bacteria, white and red blood cells, epithelial cells and crystals were identified and counted, and the samples were classified as negative or positive according to clinical cutoff values. Taking the CPT as reference, we processed 180 urine samples and compared the results with those of urine flow cytometry (UFC). Using standard evaluation metrics for our screening test, we found a similar performance for DHM and UFC, indicating DHM as a suitable and fast screening technique retaining several advantages. As a benefit of DHM, the technique is label-free and does not require sample preparation. Moreover, the phase and amplitude images of the cells and other particles present in urine are digitally recorded and can serve for further investigation afterwards.
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spelling doaj.art-2ff975eff3d64600b2deae9e4681ebd92023-11-19T00:25:13ZengMDPI AGBiosensors2079-63742023-08-0113878910.3390/bios13080789Label-Free Analysis of Urine Samples with In-Flow Digital Holographic MicroscopyLucia Gigli0Nicoletta Braidotti1Maria Augusta do R. B. F. Lima2Catalin Dacian Ciubotaru3Dan Cojoc4Alifax s.r.l. Via Merano, 30, Nimis, 33045 Udine, ItalyConsiglio Nazionale Delle Ricerche (CNR), Istituto Officina dei Materiali (IOM), Area Science Park-Basovizza, Strada Statale 14, Km 163,5, 34149 Trieste, ItalyConsiglio Nazionale Delle Ricerche (CNR), Istituto Officina dei Materiali (IOM), Area Science Park-Basovizza, Strada Statale 14, Km 163,5, 34149 Trieste, ItalyConsiglio Nazionale Delle Ricerche (CNR), Istituto Officina dei Materiali (IOM), Area Science Park-Basovizza, Strada Statale 14, Km 163,5, 34149 Trieste, ItalyConsiglio Nazionale Delle Ricerche (CNR), Istituto Officina dei Materiali (IOM), Area Science Park-Basovizza, Strada Statale 14, Km 163,5, 34149 Trieste, ItalyUrinary tract infections are among the most frequent infectious diseases and require screening a great amount of urine samples from patients. However, a high percentage of samples result as negative after urine culture plate tests (CPTs), demanding a simple and fast preliminary technique to screen out the negative samples. We propose a digital holographic microscopy (DHM) method to inspect fresh urine samples flowing in a glass capillary for 3 min, recording holograms at 2 frames per second. After digital reconstruction, bacteria, white and red blood cells, epithelial cells and crystals were identified and counted, and the samples were classified as negative or positive according to clinical cutoff values. Taking the CPT as reference, we processed 180 urine samples and compared the results with those of urine flow cytometry (UFC). Using standard evaluation metrics for our screening test, we found a similar performance for DHM and UFC, indicating DHM as a suitable and fast screening technique retaining several advantages. As a benefit of DHM, the technique is label-free and does not require sample preparation. Moreover, the phase and amplitude images of the cells and other particles present in urine are digitally recorded and can serve for further investigation afterwards.https://www.mdpi.com/2079-6374/13/8/789urine analysisdigital holographic microscopybacteria detectionurinary tract infectionsample screening
spellingShingle Lucia Gigli
Nicoletta Braidotti
Maria Augusta do R. B. F. Lima
Catalin Dacian Ciubotaru
Dan Cojoc
Label-Free Analysis of Urine Samples with In-Flow Digital Holographic Microscopy
Biosensors
urine analysis
digital holographic microscopy
bacteria detection
urinary tract infection
sample screening
title Label-Free Analysis of Urine Samples with In-Flow Digital Holographic Microscopy
title_full Label-Free Analysis of Urine Samples with In-Flow Digital Holographic Microscopy
title_fullStr Label-Free Analysis of Urine Samples with In-Flow Digital Holographic Microscopy
title_full_unstemmed Label-Free Analysis of Urine Samples with In-Flow Digital Holographic Microscopy
title_short Label-Free Analysis of Urine Samples with In-Flow Digital Holographic Microscopy
title_sort label free analysis of urine samples with in flow digital holographic microscopy
topic urine analysis
digital holographic microscopy
bacteria detection
urinary tract infection
sample screening
url https://www.mdpi.com/2079-6374/13/8/789
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