Abstract

Measuring alterations in bacteria upon antibiotic application is important for basic studies in microbiology, drug discovery, clinical diagnosis, and disease treatment. However, imaging and 3D time-lapse response analysis of individual bacteria upon antibiotic application remain largely unexplored mainly due to limitations in imaging techniques. Here, we present a method to systematically investigate the alterations in individual bacteria in 3D and quantitatively analyze the effects of antibiotics. Using optical diffraction tomography, in-situ responses of Escherichia coli and Bacillus subtilis to various concentrations of ampicillin were investigated in a label-free and quantitative manner. The presented method reconstructs the dynamic changes in the 3D refractive-index distributions of living bacteria in response to antibiotics at sub-micrometer spatial resolution.

© 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

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References

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2019 (3)

O. J. Pambos and A. N. Kapanidis, “Tracking antibiotic mechanisms,” Nat. Rev. Microbiol. 17(4), 201 (2019).
[Crossref]

S. E. Koo, S. Jang, Y. Park, and C. J. Park, “Reconstructed Three-Dimensional Images and Parameters of Individual Erythrocytes Using Optical Diffraction Tomography Microscopy,” Ann. Lab. Med. 39(2), 223–226 (2019).
[Crossref]

C. Cho, K. Nam, Y. H. Seo, K. Kim, Y. Park, J.-I. Han, and J.-Y. Lee, “Study of Optical Configurations for Multiple Enhancement of Microalgal Biomass Production,” Sci. Rep. 9(1), 1723 (2019).
[Crossref]

2018 (7)

T. Tougan, J. R. Edula, E. Takashima, M. Morita, M. Shinohara, A. Shinohara, T. Tsuboi, and T. Horii, “Molecular camouflage of Plasmodium falciparum merozoites by binding of host vitronectin to P47 fragment of SERA5,” Sci. Rep. 8(1), 5052 (2018).
[Crossref]

C. Park, S. Lee, G. Kim, S. Lee, J. Lee, T. Heo, Y. Park, and Y. Park, “Three-dimensional refractive-index distributions of individual angiosperm pollen grains,” Curr. Opt. Photonics 2(5), 460–467 (2018).

S. Kwon, Y. Lee, Y. Jung, J. H. Kim, B. Baek, B. Lim, J. Lee, I. Kim, and J. Lee, “Mitochondria-targeting indolizino [3, 2-c] quinolines as novel class of photosensitizers for photodynamic anticancer activity,” Eur. J. Med. Chem. 148, 116–127 (2018).
[Crossref]

M. Veses Garcia, H. Antypas, S. Löffler, A. Brauner, H. Andersson-Svahn, and A. Richter-Dahlfors, “Rapid Phenotypic Antibiotic Susceptibility Testing of Uropathogens Using Optical Signal Analysis on the Nanowell Slide,” Front. Microbiol. 9, 1530 (2018).
[Crossref]

Y. Park, C. Depeursinge, and G. Popescu, “Quantitative phase imaging in biomedicine,” Nat. Photonics 12(10), 578–589 (2018).
[Crossref]

C. Park, S. Shin, and Y. Park, “Generalized quantification of three-dimensional resolution in optical diffraction tomography using the projection of maximal spatial bandwidths,” J. Opt. Soc. Am. A 35(11), 1891–1898 (2018).
[Crossref]

J. Jung, S.-J. Hong, H.-B. Kim, G. Kim, M. Lee, S. Shin, S. Lee, D.-J. Kim, C.-G. Lee, and Y. Park, “Label-free non-invasive quantitative measurement of lipid contents in individual microalgal cells using refractive index tomography,” Sci. Rep. 8(1), 6524 (2018).
[Crossref]

2017 (3)

H. Alanazi, A. Canul, A. Garman, J. Quimby, and A. Vasdekis, “Robust microbial cell segmentation by optical-phase thresholding with minimal processing requirements,” Cytometry, Part A 91(5), 443–449 (2017).
[Crossref]

S. A. Yang, J. Yoon, K. Kim, and Y. Park, “Measurements of morphological and biophysical alterations in individual neuron cells associated with early neurotoxic effects in Parkinson's disease,” Cytometry, Part A 91(5), 510–518 (2017).
[Crossref]

K. Lee, K. Kim, G. Kim, S. Shin, and Y. Park, “Time-multiplexed structured illumination using a DMD for optical diffraction tomography,” Opt. Lett. 42(5), 999–1002 (2017).
[Crossref]

2016 (1)

C. Malmberg, P. Yuen, J. Spaak, O. Cars, T. Tängdén, and P. Lagerbäck, “A novel microfluidic assay for rapid phenotypic antibiotic susceptibility testing of bacteria detected in clinical blood cultures,” PLoS One 11(12), e0167356 (2016).
[Crossref]

2015 (6)

A. Jesacher, M. Ritsch-Marte, and R. Piestun, “Three-dimensional information from two-dimensional scans: a scanning microscope with postacquisition refocusing capability,” Optica 2(3), 210–213 (2015).
[Crossref]

M. Fredborg, F. Rosenvinge, E. Spillum, S. Kroghsbo, M. Wang, and T. Sondergaard, “Rapid antimicrobial susceptibility testing of clinical isolates by digital time-lapse microscopy,” Eur. J. Clin. Microbiol. Infect. Dis. 34(12), 2385–2394 (2015).
[Crossref]

S. Shin, K. Kim, J. Yoon, and Y. Park, “Active illumination using a digital micromirror device for quantitative phase imaging,” Opt. Lett. 40(22), 5407–5410 (2015).
[Crossref]

H. Park, S.-H. Hong, K. Kim, S.-H. Cho, W.-J. Lee, Y. Kim, S.-E. Lee, and Y. Park, “Characterizations of individual mouse red blood cells parasitized by Babesia microti using 3-D holographic microscopy,” Sci. Rep. 5(1), 10827 (2015).
[Crossref]

J. Lim, K. Lee, K. H. Jin, S. Shin, S. Lee, Y. Park, and J. C. Ye, “Comparative study of iterative reconstruction algorithms for missing cone problems in optical diffraction tomography,” Opt. Express 23(13), 16933–16948 (2015).
[Crossref]

Y. Jo, J. Jung, J. W. Lee, D. Shin, H. Park, K. T. Nam, J.-H. Park, and Y. Park, “Angle-resolved light scattering of individual rod-shaped bacteria based on Fourier transform light scattering,” Sci. Rep. 4(1), 5090 (2015).
[Crossref]

2014 (3)

H. Cho, T. Uehara, and T. G. Bernhardt, “Beta-lactam antibiotics induce a lethal malfunctioning of the bacterial cell wall synthesis machinery,” Cell 159(6), 1300–1311 (2014).
[Crossref]

J. Choi, J. Yoo, M. Lee, E.-G. Kim, J. S. Lee, S. Lee, S. Joo, S. H. Song, E.-C. Kim, and J. C. Lee, “A rapid antimicrobial susceptibility test based on single-cell morphological analysis,” Sci. Transl. Med. 6(267), 267ra174 (2014).
[Crossref]

C. L. Lewis, C. C. Craig, and A. G. Senecal, “Mass and density measurements of live and dead gram-negative and gram-positive bacterial populations,” Appl. Environ. Microbiol. 80(12), 3622–3631 (2014).
[Crossref]

2013 (4)

F. F. Delgado, N. Cermak, V. C. Hecht, S. Son, Y. Li, S. M. Knudsen, S. Olcum, J. M. Higgins, J. Chen, and W. H. Grover, “Intracellular water exchange for measuring the dry mass, water mass and changes in chemical composition of living cells,” PLoS One 8(7), e67590 (2013).
[Crossref]

M. Fredborg, K. R. Andersen, E. Jørgensen, A. Droce, T. Olesen, B. B. Jensen, F. S. Rosenvinge, and T. E. Sondergaard, “Real-time optical antimicrobial susceptibility testing,” J. Clin. Microbiol. 51(7), 2047–2053 (2013).
[Crossref]

K. Kim, H. Yoon, M. Diez-Silva, M. Dao, R. R. Dasari, and Y. Park, “High-resolution three-dimensional imaging of red blood cells parasitized by Plasmodium falciparum and in situ hemozoin crystals using optical diffraction tomography,” J. Biomed. Opt. 19(1), 011005 (2013).
[Crossref]

B. Kemper, Á. Barroso, M. Woerdemann, L. Dewenter, A. Vollmer, R. Schubert, A. Mellmann, G. von Bally, and C. Denz, “Towards 3D modelling and imaging of infection scenarios at the single cell level using holographic optical tweezers and digital holographic microscopy,” J. Biophotonics 6(3), 260–266 (2013).
[Crossref]

2012 (4)

E. Efstratiou, A. I. Hussain, P. S. Nigam, J. E. Moore, M. A. Ayub, and J. R. Rao, “Antimicrobial activity of Calendula officinalis petal extracts against fungi, as well as Gram-negative and Gram-positive clinical pathogens,” Complement. Ther. Clin. Prac. 18(3), 173–176 (2012).
[Crossref]

U. Neugebauer, C. Große, M. Bauer, B. Kemper, A. Barroso-Pena, A. Bauwens, M. Glueder, M. Woerdemann, L. Dewenter, and C. Denz, “From Infection to Detection: Imaging S. aureus–host interactions,” Biomed. Tech. 57(SI-1 Track-B), 503–506 (2012).
[Crossref]

Z. Yao, D. Kahne, and R. Kishony, “Distinct single-cell morphological dynamics under beta-lactam antibiotics,” Mol. Cell 48(5), 705–712 (2012).
[Crossref]

E. C. Jensen, “Use of fluorescent probes: their effect on cell biology and limitations,” Anat. Rec. 295(12), 2031–2036 (2012).
[Crossref]

2011 (1)

2010 (4)

R. I. Aminov, “A brief history of the antibiotic era: lessons learned and challenges for the future,” Front. Microbiol. 1, 134 (2010).
[Crossref]

M. A. Kohanski, D. J. Dwyer, and J. J. Collins, “How antibiotics kill bacteria: from targets to networks,” Nat. Rev. Microbiol. 8(6), 423–435 (2010).
[Crossref]

K. F. Kong, L. Schneper, and K. Mathee, “Beta-lactam antibiotics: from antibiosis to resistance and bacteriology,” APMIS 118(1), 1–36 (2010).
[Crossref]

S. Tanaka, M. R. Sawaya, and T. O. Yeates, “Structure and mechanisms of a protein-based organelle in Escherichia coli,” Science 327(5961), 81–84 (2010).
[Crossref]

2009 (1)

2008 (3)

B. R. Bochner, L. Giovannetti, and C. Viti, “Important discoveries from analysing bacterial phenotypes,” Mol. Microbiol. 70(2), 274–280 (2008).
[Crossref]

G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell. Physiol. 295(2), C538–C544 (2008).
[Crossref]

J. B. Parsons, S. D. Dinesh, E. Deery, H. K. Leech, A. A. Brindley, D. Heldt, S. Frank, C. M. Smales, H. Lünsdorf, and A. Rambach, “Biochemical and structural insights into bacterial organelle form and biogenesis,” J. Biol. Chem. 283(21), 14366–14375 (2008).
[Crossref]

2005 (2)

J. Conly and B. Johnston, “Where are all the new antibiotics? The new antibiotic paradox,” Can. J. Infect. Dis. Med. Microbiol. 16(3), 159–160 (2005).
[Crossref]

K. El Kirat, I. Burton, V. Dupres, and Y. Dufrene, “Sample preparation procedures for biological atomic force microscopy,” J. Microsc. 218(3), 199–207 (2005).
[Crossref]

2002 (1)

V. Lauer, “New approach to optical diffraction tomography yielding a vector equation of diffraction tomography and a novel tomographic microscope,” J. Microsc. 205(2), 165–176 (2002).
[Crossref]

1993 (1)

W. D. Donachie, “The cell cycle of Escherichia coli,” Annu. Rev. Microbiol. 47(1), 199–230 (1993).
[Crossref]

1992 (1)

H. C. Neu, “The crisis in antibiotic resistance,” Science 257(5073), 1064–1073 (1992).
[Crossref]

1991 (1)

A. G. Marr, “Growth rate of Escherichia coli,” Microbiol. Mol. Biol. R. 55, 316–333 (1991).

1982 (1)

1981 (1)

1975 (2)

B. G. Spratt, “Distinct penicillin binding proteins involved in the division, elongation, and shape of Escherichia coli K12,” Proc. Natl. Acad. Sci. 72(8), 2999–3003 (1975).
[Crossref]

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H. Alanazi, A. Canul, A. Garman, J. Quimby, and A. Vasdekis, “Robust microbial cell segmentation by optical-phase thresholding with minimal processing requirements,” Cytometry, Part A 91(5), 443–449 (2017).
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R. Gerchberg, “Super-resolution through error energy reduction,” Opt. Acta 21(9), 709–720 (1974).
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B. R. Bochner, L. Giovannetti, and C. Viti, “Important discoveries from analysing bacterial phenotypes,” Mol. Microbiol. 70(2), 274–280 (2008).
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S. E. Koo, S. Jang, Y. Park, and C. J. Park, “Reconstructed Three-Dimensional Images and Parameters of Individual Erythrocytes Using Optical Diffraction Tomography Microscopy,” Ann. Lab. Med. 39(2), 223–226 (2019).
[Crossref]

Park, H.

Y. Jo, J. Jung, J. W. Lee, D. Shin, H. Park, K. T. Nam, J.-H. Park, and Y. Park, “Angle-resolved light scattering of individual rod-shaped bacteria based on Fourier transform light scattering,” Sci. Rep. 4(1), 5090 (2015).
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H. Park, S.-H. Hong, K. Kim, S.-H. Cho, W.-J. Lee, Y. Kim, S.-E. Lee, and Y. Park, “Characterizations of individual mouse red blood cells parasitized by Babesia microti using 3-D holographic microscopy,” Sci. Rep. 5(1), 10827 (2015).
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Park, J.-H.

Y. Jo, J. Jung, J. W. Lee, D. Shin, H. Park, K. T. Nam, J.-H. Park, and Y. Park, “Angle-resolved light scattering of individual rod-shaped bacteria based on Fourier transform light scattering,” Sci. Rep. 4(1), 5090 (2015).
[Crossref]

Park, Y.

S. E. Koo, S. Jang, Y. Park, and C. J. Park, “Reconstructed Three-Dimensional Images and Parameters of Individual Erythrocytes Using Optical Diffraction Tomography Microscopy,” Ann. Lab. Med. 39(2), 223–226 (2019).
[Crossref]

C. Cho, K. Nam, Y. H. Seo, K. Kim, Y. Park, J.-I. Han, and J.-Y. Lee, “Study of Optical Configurations for Multiple Enhancement of Microalgal Biomass Production,” Sci. Rep. 9(1), 1723 (2019).
[Crossref]

Y. Park, C. Depeursinge, and G. Popescu, “Quantitative phase imaging in biomedicine,” Nat. Photonics 12(10), 578–589 (2018).
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C. Park, S. Lee, G. Kim, S. Lee, J. Lee, T. Heo, Y. Park, and Y. Park, “Three-dimensional refractive-index distributions of individual angiosperm pollen grains,” Curr. Opt. Photonics 2(5), 460–467 (2018).

C. Park, S. Lee, G. Kim, S. Lee, J. Lee, T. Heo, Y. Park, and Y. Park, “Three-dimensional refractive-index distributions of individual angiosperm pollen grains,” Curr. Opt. Photonics 2(5), 460–467 (2018).

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J. Jung, S.-J. Hong, H.-B. Kim, G. Kim, M. Lee, S. Shin, S. Lee, D.-J. Kim, C.-G. Lee, and Y. Park, “Label-free non-invasive quantitative measurement of lipid contents in individual microalgal cells using refractive index tomography,” Sci. Rep. 8(1), 6524 (2018).
[Crossref]

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[Crossref]

H. Park, S.-H. Hong, K. Kim, S.-H. Cho, W.-J. Lee, Y. Kim, S.-E. Lee, and Y. Park, “Characterizations of individual mouse red blood cells parasitized by Babesia microti using 3-D holographic microscopy,” Sci. Rep. 5(1), 10827 (2015).
[Crossref]

S. Shin, K. Kim, J. Yoon, and Y. Park, “Active illumination using a digital micromirror device for quantitative phase imaging,” Opt. Lett. 40(22), 5407–5410 (2015).
[Crossref]

J. Lim, K. Lee, K. H. Jin, S. Shin, S. Lee, Y. Park, and J. C. Ye, “Comparative study of iterative reconstruction algorithms for missing cone problems in optical diffraction tomography,” Opt. Express 23(13), 16933–16948 (2015).
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Y. Jo, J. Jung, J. W. Lee, D. Shin, H. Park, K. T. Nam, J.-H. Park, and Y. Park, “Angle-resolved light scattering of individual rod-shaped bacteria based on Fourier transform light scattering,” Sci. Rep. 4(1), 5090 (2015).
[Crossref]

K. Kim, H. Yoon, M. Diez-Silva, M. Dao, R. R. Dasari, and Y. Park, “High-resolution three-dimensional imaging of red blood cells parasitized by Plasmodium falciparum and in situ hemozoin crystals using optical diffraction tomography,” J. Biomed. Opt. 19(1), 011005 (2013).
[Crossref]

S. K. Debnath and Y. Park, “Real-time quantitative phase imaging with a spatial phase-shifting algorithm,” Opt. Lett. 36(23), 4677–4679 (2011).
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G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell. Physiol. 295(2), C538–C544 (2008).
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M. Lee, Y.-H. Lee, J. Song, G. Kim, Y. Jo, H. Min, C. H. Kim, and Y. Park, “Deep-learning based three-dimensional label-free tracking and analysis of immunological synapses of chimeric antigen receptor T cells,” BioRxiv539858 (2019).

K. Kim, J. Yoon, S. Shin, S. Lee, S.-A. Yang, and Y. Park, “Optical diffraction tomography techniques for the study of cell pathophysiology,” J. Biomed. Photonics Eng.020201-1–020201-16 (2016).

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G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell. Physiol. 295(2), C538–C544 (2008).
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G. Kim, D. Ahn, M. Kang, Y. Jo, D. Ryu, H. Kim, J. Song, J. S. Ryu, G. Choi, and H. J. Chung, “Rapid and label-free identification of individual bacterial pathogens exploiting three-dimensional quantitative phase imaging and deep learning,” BioRxiv, 596486 (2019).

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C. Cho, K. Nam, Y. H. Seo, K. Kim, Y. Park, J.-I. Han, and J.-Y. Lee, “Study of Optical Configurations for Multiple Enhancement of Microalgal Biomass Production,” Sci. Rep. 9(1), 1723 (2019).
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Y. Jo, J. Jung, J. W. Lee, D. Shin, H. Park, K. T. Nam, J.-H. Park, and Y. Park, “Angle-resolved light scattering of individual rod-shaped bacteria based on Fourier transform light scattering,” Sci. Rep. 4(1), 5090 (2015).
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G. Kim, D. Ahn, M. Kang, Y. Jo, D. Ryu, H. Kim, J. Song, J. S. Ryu, G. Choi, and H. J. Chung, “Rapid and label-free identification of individual bacterial pathogens exploiting three-dimensional quantitative phase imaging and deep learning,” BioRxiv, 596486 (2019).

Song, S. H.

J. Choi, J. Yoo, M. Lee, E.-G. Kim, J. S. Lee, S. Lee, S. Joo, S. H. Song, E.-C. Kim, and J. C. Lee, “A rapid antimicrobial susceptibility test based on single-cell morphological analysis,” Sci. Transl. Med. 6(267), 267ra174 (2014).
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M. Fredborg, F. Rosenvinge, E. Spillum, S. Kroghsbo, M. Wang, and T. Sondergaard, “Rapid antimicrobial susceptibility testing of clinical isolates by digital time-lapse microscopy,” Eur. J. Clin. Microbiol. Infect. Dis. 34(12), 2385–2394 (2015).
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C. Qi, C. W. Stratton, and X. Zheng, “Phenotypic testing of bacterial antimicrobial susceptibility,” in Advanced Techniques in Diagnostic Microbiology (Springer, 2006), pp. 63–83.

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Tabin, C. J.

S. Oh, C. Lee, D. Fu, W. Yang, A. Li, C. Ran, W. Yin, C. J. Tabin, X. S. Xie, and M. W. Kirschner, “In situ measurement of absolute concentrations by Normalized Raman Imaging,” BioRxiv, 629543 (2019).

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S. Tanaka, M. R. Sawaya, and T. O. Yeates, “Structure and mechanisms of a protein-based organelle in Escherichia coli,” Science 327(5961), 81–84 (2010).
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C. Malmberg, P. Yuen, J. Spaak, O. Cars, T. Tängdén, and P. Lagerbäck, “A novel microfluidic assay for rapid phenotypic antibiotic susceptibility testing of bacteria detected in clinical blood cultures,” PLoS One 11(12), e0167356 (2016).
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T. Tougan, J. R. Edula, E. Takashima, M. Morita, M. Shinohara, A. Shinohara, T. Tsuboi, and T. Horii, “Molecular camouflage of Plasmodium falciparum merozoites by binding of host vitronectin to P47 fragment of SERA5,” Sci. Rep. 8(1), 5052 (2018).
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M. Veses Garcia, H. Antypas, S. Löffler, A. Brauner, H. Andersson-Svahn, and A. Richter-Dahlfors, “Rapid Phenotypic Antibiotic Susceptibility Testing of Uropathogens Using Optical Signal Analysis on the Nanowell Slide,” Front. Microbiol. 9, 1530 (2018).
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B. Kemper, Á. Barroso, M. Woerdemann, L. Dewenter, A. Vollmer, R. Schubert, A. Mellmann, G. von Bally, and C. Denz, “Towards 3D modelling and imaging of infection scenarios at the single cell level using holographic optical tweezers and digital holographic microscopy,” J. Biophotonics 6(3), 260–266 (2013).
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Yang, S. A.

S. A. Yang, J. Yoon, K. Kim, and Y. Park, “Measurements of morphological and biophysical alterations in individual neuron cells associated with early neurotoxic effects in Parkinson's disease,” Cytometry, Part A 91(5), 510–518 (2017).
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Yang, W.

S. Oh, C. Lee, D. Fu, W. Yang, A. Li, C. Ran, W. Yin, C. J. Tabin, X. S. Xie, and M. W. Kirschner, “In situ measurement of absolute concentrations by Normalized Raman Imaging,” BioRxiv, 629543 (2019).

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S. Oh, C. Lee, D. Fu, W. Yang, A. Li, C. Ran, W. Yin, C. J. Tabin, X. S. Xie, and M. W. Kirschner, “In situ measurement of absolute concentrations by Normalized Raman Imaging,” BioRxiv, 629543 (2019).

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J. Choi, J. Yoo, M. Lee, E.-G. Kim, J. S. Lee, S. Lee, S. Joo, S. H. Song, E.-C. Kim, and J. C. Lee, “A rapid antimicrobial susceptibility test based on single-cell morphological analysis,” Sci. Transl. Med. 6(267), 267ra174 (2014).
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K. Kim, H. Yoon, M. Diez-Silva, M. Dao, R. R. Dasari, and Y. Park, “High-resolution three-dimensional imaging of red blood cells parasitized by Plasmodium falciparum and in situ hemozoin crystals using optical diffraction tomography,” J. Biomed. Opt. 19(1), 011005 (2013).
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Yoon, J.

S. A. Yang, J. Yoon, K. Kim, and Y. Park, “Measurements of morphological and biophysical alterations in individual neuron cells associated with early neurotoxic effects in Parkinson's disease,” Cytometry, Part A 91(5), 510–518 (2017).
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S. Shin, K. Kim, J. Yoon, and Y. Park, “Active illumination using a digital micromirror device for quantitative phase imaging,” Opt. Lett. 40(22), 5407–5410 (2015).
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K. Kim, J. Yoon, S. Shin, S. Lee, S.-A. Yang, and Y. Park, “Optical diffraction tomography techniques for the study of cell pathophysiology,” J. Biomed. Photonics Eng.020201-1–020201-16 (2016).

Yuen, P.

C. Malmberg, P. Yuen, J. Spaak, O. Cars, T. Tängdén, and P. Lagerbäck, “A novel microfluidic assay for rapid phenotypic antibiotic susceptibility testing of bacteria detected in clinical blood cultures,” PLoS One 11(12), e0167356 (2016).
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Am. J. Physiol. Cell. Physiol. (1)

G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell. Physiol. 295(2), C538–C544 (2008).
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Ann. Lab. Med. (1)

S. E. Koo, S. Jang, Y. Park, and C. J. Park, “Reconstructed Three-Dimensional Images and Parameters of Individual Erythrocytes Using Optical Diffraction Tomography Microscopy,” Ann. Lab. Med. 39(2), 223–226 (2019).
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APMIS (1)

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Appl. Environ. Microbiol. (1)

C. L. Lewis, C. C. Craig, and A. G. Senecal, “Mass and density measurements of live and dead gram-negative and gram-positive bacterial populations,” Appl. Environ. Microbiol. 80(12), 3622–3631 (2014).
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Biomed. Tech. (1)

U. Neugebauer, C. Große, M. Bauer, B. Kemper, A. Barroso-Pena, A. Bauwens, M. Glueder, M. Woerdemann, L. Dewenter, and C. Denz, “From Infection to Detection: Imaging S. aureus–host interactions,” Biomed. Tech. 57(SI-1 Track-B), 503–506 (2012).
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Cell (1)

H. Cho, T. Uehara, and T. G. Bernhardt, “Beta-lactam antibiotics induce a lethal malfunctioning of the bacterial cell wall synthesis machinery,” Cell 159(6), 1300–1311 (2014).
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Complement. Ther. Clin. Prac. (1)

E. Efstratiou, A. I. Hussain, P. S. Nigam, J. E. Moore, M. A. Ayub, and J. R. Rao, “Antimicrobial activity of Calendula officinalis petal extracts against fungi, as well as Gram-negative and Gram-positive clinical pathogens,” Complement. Ther. Clin. Prac. 18(3), 173–176 (2012).
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Curr. Opt. Photonics (1)

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Figures (5)

Fig. 1.
Fig. 1. Measurements of the 3D RI distribution. (a) Experimental setup. M1−M4, mirror; L1−L5, lens; FC, fiber coupler; DMD, digital micromirror device. (b) Sequential illumination scanning for bacteria at 71 illumination angles and the corresponding 71 holograms. (c) Images of the retrieved amplitudes and phases corresponding to the holograms. (d) Three-section view of the reconstructed RI distribution. (e) 3D rendered image of bacteria. The color map shows various colors depending on the value of RI and its gradient value.
Fig. 2.
Fig. 2. (a) Three-section views of the reconstructed RI distributions of E. coli without antibiotics over time (at 0, 120, and 240 minutes). (b) 3D rendered images of bacteria corresponding to (a). The color map shows various colors depending on the value of the RI. (c) The number of E. coli for the control group over time and the fitted curve. R-square = 0.9741.
Fig. 3.
Fig. 3. Maximum RI projection images with ampicillin over time. The red arrows indicate a bulge formed in E. coli. The entry with a red border is presumed to be near the point at which the bacteria die. (a) and (b) are E. coli and B. subtilis, respectively.
Fig. 4.
Fig. 4. Cell volume, cellular dry mass, cytoplasm concentration and corresponding RI depending on the concentrations of ampicillin (0, 20, 100, and 300 µg/mL) and time at the maximum volume. (a−d) and (e−h) are E. coli and B. subtilis, respectively. The number of dataset for E. coli (control, 20, 100, and 300 µg/mL) and B. subtilis (control, 20, 100, and 300 µg/mL) is 4, 5, 5, 4, 4, 4, 5, and 5, respectively.
Fig. 5.
Fig. 5. (a−c) The relative value of cell volume for E. coli and B. subtilis with the ampicillin concentration of (a) 20 µg/mL (b) 100 µg/mL (c) 300 µg/mL. (d) The relative value of the maximum volume for E. coli and B. subtilis. The number of dataset for E. coli (20, 100, and 300 µg/mL) and B. subtilis (20, 100, and 300 µg/mL) is 5, 5, 4, 4, 5, and 5, respectively.

Equations (1)

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( 2 + k i 2 ) u s ( r ) = u ( r ) f ( r ) ,