Rapid Detection and Analysis of Raman Spectra of Bacteria in Multiple Fields of View Based on Image Stitching Technique

Background: Due to antibiotic abuse, the problem of bacterial resistance is becoming increasingly serious, and rapid detection of bacterial resistance has become an urgent issue. Because under the action of antibiotics, different active bacteria have different metabolism of heavy water, antibiotic r...

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Main Authors: Xiaohui Dou, Fengna Yang, Nan Wang, Ying Xue, Haoran Hu, Bei Li
Format: Article
Language:English
Published: IMR Press 2023-10-01
Series:Frontiers in Bioscience-Landmark
Subjects:
Online Access:https://www.imrpress.com/journal/FBL/28/10/10.31083/j.fbl2810249
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author Xiaohui Dou
Fengna Yang
Nan Wang
Ying Xue
Haoran Hu
Bei Li
author_facet Xiaohui Dou
Fengna Yang
Nan Wang
Ying Xue
Haoran Hu
Bei Li
author_sort Xiaohui Dou
collection DOAJ
description Background: Due to antibiotic abuse, the problem of bacterial resistance is becoming increasingly serious, and rapid detection of bacterial resistance has become an urgent issue. Because under the action of antibiotics, different active bacteria have different metabolism of heavy water, antibiotic resistance of bacteria can be identified according to the existence of a C-D peak in the 2030–2400 cm-1 range in the Raman spectrum. Methods: To ensure data veracity, a large number of bacteria need to be detected, however, due to the limitation of the field of view of the high magnification objective, the number of single cells in a single field of view is very small. By combining an image stitching algorithm, image recognition algorithm, and processing of Raman spectrum and peak-seeking algorithm, can identify and locate single cells in multiple fields of view at one time and can discriminate whether they are Antimicrobial-resistant bacteria. Results: In experiments 1 and 2, 2706 bacteria in 9 × 11 fields of view and 2048 bacteria in 11 × 11 fields of view were detected. Results showed that in experiment 1, there are 1137 antibiotic-resistant bacteria, accounting for 42%, and 1569 sensitive bacteria, accounting for 58%. In experiment 2, there are 1087 antibiotic-resistant bacteria, accounting for 53%, and 961 sensitive bacteria, accounting for 47%. It showed excellent performance in terms of speed and recognition accuracy as compared to traditional manual detection approaches. And solves the problems of low accuracy of data, a large number of manual experiments, and low efficiency due to the small number of single cells in the high magnification field of view and different peak-seeking parameters of different Raman spectra. Conclusions: The detection and analysis method of bacterial Raman spectra based on image stitching can be used for unattended, automatic, rapid and accurate detection of single cells at high magnification with multiple fields of view. With the characteristics of automatic, high-throughput, rapid, and accurate identification, it can be used as an unattended, universal and non-invasive means to measure antibiotic-resistant bacteria to screen for effective antibiotics, which is of great importance for studying the persistence and spread of antibiotics in bacterial pathogens.
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spelling doaj.art-e412a11a647b4eac969697dd06f94d012023-11-03T02:31:25ZengIMR PressFrontiers in Bioscience-Landmark2768-67012023-10-01281024910.31083/j.fbl2810249S2768-6701(23)00930-9Rapid Detection and Analysis of Raman Spectra of Bacteria in Multiple Fields of View Based on Image Stitching TechniqueXiaohui Dou0Fengna Yang1Nan Wang2Ying Xue3Haoran Hu4Bei Li5Department of Biomedical Engineering, Wenzhou Medical University, 325035 Wenzhou, Zhejiang, ChinaDepartment of Biomedical Engineering, Wenzhou Medical University, 325035 Wenzhou, Zhejiang, ChinaDepartment of Research and Development, Hooke Instruments, 130033 Changchun, Jilin, ChinaDepartment of Research and Development, Hooke Instruments, 130033 Changchun, Jilin, ChinaDepartment of Biomedical Engineering, Wenzhou Medical University, 325035 Wenzhou, Zhejiang, ChinaDepartment of Biomedical Engineering, Wenzhou Medical University, 325035 Wenzhou, Zhejiang, ChinaBackground: Due to antibiotic abuse, the problem of bacterial resistance is becoming increasingly serious, and rapid detection of bacterial resistance has become an urgent issue. Because under the action of antibiotics, different active bacteria have different metabolism of heavy water, antibiotic resistance of bacteria can be identified according to the existence of a C-D peak in the 2030–2400 cm-1 range in the Raman spectrum. Methods: To ensure data veracity, a large number of bacteria need to be detected, however, due to the limitation of the field of view of the high magnification objective, the number of single cells in a single field of view is very small. By combining an image stitching algorithm, image recognition algorithm, and processing of Raman spectrum and peak-seeking algorithm, can identify and locate single cells in multiple fields of view at one time and can discriminate whether they are Antimicrobial-resistant bacteria. Results: In experiments 1 and 2, 2706 bacteria in 9 × 11 fields of view and 2048 bacteria in 11 × 11 fields of view were detected. Results showed that in experiment 1, there are 1137 antibiotic-resistant bacteria, accounting for 42%, and 1569 sensitive bacteria, accounting for 58%. In experiment 2, there are 1087 antibiotic-resistant bacteria, accounting for 53%, and 961 sensitive bacteria, accounting for 47%. It showed excellent performance in terms of speed and recognition accuracy as compared to traditional manual detection approaches. And solves the problems of low accuracy of data, a large number of manual experiments, and low efficiency due to the small number of single cells in the high magnification field of view and different peak-seeking parameters of different Raman spectra. Conclusions: The detection and analysis method of bacterial Raman spectra based on image stitching can be used for unattended, automatic, rapid and accurate detection of single cells at high magnification with multiple fields of view. With the characteristics of automatic, high-throughput, rapid, and accurate identification, it can be used as an unattended, universal and non-invasive means to measure antibiotic-resistant bacteria to screen for effective antibiotics, which is of great importance for studying the persistence and spread of antibiotics in bacterial pathogens.https://www.imrpress.com/journal/FBL/28/10/10.31083/j.fbl2810249image stitchingraman spectrumimage recognitionantibiotic resistancemultiple fields of viewheavy water labeling
spellingShingle Xiaohui Dou
Fengna Yang
Nan Wang
Ying Xue
Haoran Hu
Bei Li
Rapid Detection and Analysis of Raman Spectra of Bacteria in Multiple Fields of View Based on Image Stitching Technique
Frontiers in Bioscience-Landmark
image stitching
raman spectrum
image recognition
antibiotic resistance
multiple fields of view
heavy water labeling
title Rapid Detection and Analysis of Raman Spectra of Bacteria in Multiple Fields of View Based on Image Stitching Technique
title_full Rapid Detection and Analysis of Raman Spectra of Bacteria in Multiple Fields of View Based on Image Stitching Technique
title_fullStr Rapid Detection and Analysis of Raman Spectra of Bacteria in Multiple Fields of View Based on Image Stitching Technique
title_full_unstemmed Rapid Detection and Analysis of Raman Spectra of Bacteria in Multiple Fields of View Based on Image Stitching Technique
title_short Rapid Detection and Analysis of Raman Spectra of Bacteria in Multiple Fields of View Based on Image Stitching Technique
title_sort rapid detection and analysis of raman spectra of bacteria in multiple fields of view based on image stitching technique
topic image stitching
raman spectrum
image recognition
antibiotic resistance
multiple fields of view
heavy water labeling
url https://www.imrpress.com/journal/FBL/28/10/10.31083/j.fbl2810249
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