Identifying the microbial species present in a sample is critical in healthcare, industry, ecology and even national security. The traditional diagnosis process involves the growth of colonies of microorganisms on a solid-state medium in a Petri dish. This step provides great amounts of biological material, allows spatial separation of the different microorganisms and can help to visually discriminate potentially harmful species. However, it does not allow a precise identification and thereby requires further sample preparation and analysis to offer a proper diagnosis. Here, a new optical-based Petri dish analysis technology is discussed. This technique, called lensless multispectral mid-infrared imaging, relies on the acquisition of images at nine wavelengths corresponding to relevant chemical functions. It provides both morphological and discrete spectral data, which allows to discriminate even closely related species. The instrument is simply made of a laser source (four quantum cascade lasers) and a microbolometer array as the imager. A total of 1050 colonies belonging to three Staphylococcus species and two strains of Staphylococcus epidermidis have been acquired. After feature extraction and classification by Support Vector Machine, the tenfold cross-validation test yields a correct identification rate between 93 % and 96 %, with only a 5 % confusion between the two strains of S. epidermidis. Further work on the data analysis algorithms could dramatically improve these already promising results. Therefore, considering its label-free and non-destructive aspects, as well as its absence of secondary sample preparation, this technology has a great potential to offer precise Petri-dish based diagnosis.
Microbial identification is a critical process aiming at identifying the species contained in a biological sample, with applications in healthcare, industry or even national security. Traditionally, this process relies either on MALDI-TOF mass spectroscopy, on biochemical tests and on the observation of the morphology of colonies after growth on a Petri dish. Here is presented an innovative method for label-free optical identification of pathogens, based on the multispectral infrared imaging of colonies. This lensless imaging technique enables a high-throughput analysis and wide-field analysis of agar plates. It could yield very high correct identification rates as it relies on an optical fingerprint gathering both spectroscopic and morphologic features. The setup consists of a Quantum Cascade Lasers light source and an imager, a square 2.72 by 2.72 mm uncooled bolometer array. Microorganisms to be analyzed are streaked on a porous growth support compatible with infrared imaging, laid on top of an agar plate for incubation. When imaging is performed, growth support is put in close contact with the imaging sensor and illuminated at different wavelengths. After acquisition, an image descriptor based on spectral and morphological features is extracted for each microbial colony. Supervised classification is finally performed with a Support Vector Machine algorithm and tested with tenfold cross-validation. A first database collecting 1012 multispectral images of colonies belonging to five different species has already been acquired with this system, resulting in a correct identification rate of 92%. For these experiments, multispectral images are acquired at nine different wavelengths, between 5.6 and 8 μm. Considering the optimization possibilities of the image descriptors currently used and the ongoing development of the uncooled bolometers technology, these very first results are promising and could be dramatically improved with further experiments. Thereby, mid-infrared multispectral lensless imaging has the potential to become a fast and precise Petri dish analysis technology.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.