Direct and label-free gram classification of bacterial colonies on agar using hyperspectral imaging

Midahuen, Rony; Barlas, P; Fulchiron, Corine; Laloum, E.; Mahé, Pierre; Leroux, Denis

doi:10.1117/12.2306256

Cited by 2 publications

(3 citation statements)

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“…Optical identi cation techniques can be divided into two groups: the vibrational spectroscopies, which yield a composition-based ngerprint [2,5,6], and the morphotype-based analysis, such as plate imaging [7,8] or elastic light scattering [9][10][11]. Both approaches are widely documented, each with its own perks and drawbacks, but only a few studies offer to combine these two very different types of information : composition and morphotype.…”

Section: Introductionmentioning

confidence: 99%

“…This so-called hyperspectral imaging works either by scanning a sample point-bypoint with a spectrometer, by a line scanning ("pushbroom imaging") or by a whole-image acquisition with a focal-plane imager linked to a spectrometer [13]. Such approaches have already been employed for Gram classi cation of bacterial colonies [7], and sometimes identi cation down to species [8,14].…”

Section: Introductionmentioning

confidence: 99%

“…wavelengths between 0.4 and 1 µm). This range of wavelengths, although it allow a certain level of classi cation in some speci c cases [7],…”

Section: Introductionmentioning

confidence: 99%

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Microbial identification through multispectral infrared imaging of colonies: combining chemical and morphotype analysis

Galudec¹,

Dupoy

Rebuffel³

et al. 2023

Preprint

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Most of optical methods for the identification of microorganisms rely either on vibrational spectroscopies, such as Raman and infrared spectroscopy, or on morphotype imaging within the visible range. With the most recent developments of quantum cascade lasers, a fast imaging system able to collect both of these information, vibrational (composition-based) and morphotype, is now within reach. However, the potential of such system to be more precise than conventional imaging or spectroscopy has yet to be demonstrated. Here, we report a practical protocol for microbial colonies identification by means of QCL-based multispectral infrared imaging, as well as the acquisition of a first database for performance evaluation. To this end, optical fingerprints of 2253 colonies belonging to 8 microbial species were recorded at 9 different wavelengths in the mid-infrared range. Image descriptors and machine learning approaches allowed for classification with a 94.4% correct identification rate. Strain on three strains of S. epidermidis appeared feasible and as effective as species-level classification. Therefore, multispectral imaging in the mid-infrared has the potential to offer a powerful label-free and non-destructive technology for microbial identification, and could be used for strain typing.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Microbial identification through multispectral infrared imaging of colonies: combining chemical and morphotype analysis

Galudec¹,

Dupoy

Rebuffel³

et al. 2023

Preprint

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Optical methods for bacterial detection and characterization

et al. 2021

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Rapid identification and enumeration of bacteria are critical, given the surge of antibiotic-resistance, global exchange of food products, and the use of bacteria for bioremediation, pharmaceutical, and food production. In response, a wide range of methods are being developed that can be broadly classified as nucleic acid-based, structure-based, mass spectrometry-based, and optically based. Optical methods have generated interest given the potential for rapid, non-destructive, high-throughput, and amplification-free measurements that require minimal sample preparation. This Perspective reviews optical methods, which are applied to identification, enumeration, and greater understanding of bacteria routinely and more importantly at the cutting edge of research, with the aim of identifying gaps and opportunities for development. We have focused primarily on methods that directly measure bacteria and not their effect on the sample matrix or sensing, which requires a biorecognition element (i.e., label specific to some component of the bacterium). We identify gaps in the existing techniques and avenues for innovation. Finally, we suggest the parameters that should be considered and recorded when reporting the development of existing and new methods for bacterial characterization. This Perspective is intended for physicists interested in developing new optical methods for the study of bacteria and microbiologists in need of an optical technique for bacterial applications.

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Direct and label-free gram classification of bacterial colonies on agar using hyperspectral imaging

Cited by 2 publications

References 5 publications

Microbial identification through multispectral infrared imaging of colonies: combining chemical and morphotype analysis

Microbial identification through multispectral infrared imaging of colonies: combining chemical and morphotype analysis

Optical methods for bacterial detection and characterization

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