Imaging-in-flow: digital holographic microscopy as a novel tool to detect and classify nanoplanktonic organisms

Traditional taxonomic identification of planktonic organisms is based on light microscopy, which is both time-consuming and tedious. In response, novel ways of automated (machine) identification, such as flow cytometry, have been investigated over the last two decades. To improve the taxonomic resolution of particle analysis, recent developments have focused on "imaging-in-flow," i.e., the ability to acquire microscopic images of planktonic cells in a flow-through mode. Imaging-in-flow systems are traditionally based on classical brightfield microscopy and are faced with a number of issues that decrease the classification performance and accuracy (e. g., projection variance of cells, migration of cells out of the focus plane). Here, we demonstrate that a combination of digital holographic microscopy (DHM) with imaging-in-flow can improve the detection and classification of planktonic organisms. In addition to light intensity information, DHM provides quantitative phase information, which generates an additional and independent set of features that can be used in classification algorithms. Moreover, the capability of digitally refocusing greatly increases the depth of field, enables a more accurate focusing of cells, and reduces the effects of position variance. Nanoplanktonic organisms similar in shape were successfully classified from images captured with an off-axis DHM with partial coherence. Textural features based on DHM phase information proved more efficient in separating the three tested phytoplankton species compared with shape-based features or textural features based on light intensity. An overall classification score of 92.4% demonstrates the potential of holographic-based imaging-in-flow for similar looking organisms in the nanoplankton range

Cinétique de l'action des antibiotiques en milieu plasmatique

Doctorat en sciences médicalesPublié dans la revue "Acta Clinica Belgica", Vol. 22, Suppl. 4 (1967).info:eu-repo/semantics/nonPublishe

Automated Count and Size Evaluation of Colonies of Bacteria Grown in a Zonal Concentration Gradient of Antimicrobial Agent

The quantitative study (counting and size and surface evaluation of bacterial colonies) of the activity of an antimicrobial agent against a microbial population growing in solid medium can be performed by an electronic image analyzer. The Zeiss Micro-Videomat allowed the detection of even slight antimicrobial effects, which would be difficult to detect by colony counting alone and would escape the manual procedures of observation. The potential of the new method of investigation was illustrated by the examination of Staphylococcus aureus inhibition zones produced by disks of penicillin G and sulfadiazine

Significance of the detection of beta-galactosidase and of beta-xylosidase in the taxonomic study of the genus Haemophilus.

A total of 314 strains of Haemophilus, isolated from clinical samples, were studied for the production of beta-galactosidase and beta-xylosidase. None of the H. influenzae strains studied (9 beta-lactamase positive strains and 129 beta-lactamase negative strains) possessed these enzymes. Both enzymes were almost constantly observed among strains of H. paraphrophilus (10 strains studied) and of H. paraphrohaemolyticus (9 strains studied). Among the other species (H. parainfluenzae, 55 strains; H. haemolyticus, 5 strains; H. parahaemolyticus, 97 strains), beta-galactosidase was present in about 30% of the strains studied whereas beta-xylosidase was detected occasionally (3% of the strains studied). Detection of these two enzymes could be a valuable test for the taxonomic study of the genus Haemophilus. However, the type of substrate used for the detection of beta-xylosidase is important: use of the para-nitro-phenyl-beta-xylopyranoside yielded more positive results than the use of its ortho-isomer

Taxonomy of Acinetobacter: the usefulness of beta-D-xyloside xylohydrolase for strain differentiation.

One hundred and twenty-two clinical isolates of Acinetobacter were studied for the presence of beta-galactosidase and of beta-xylosidase, for biochemical characteristics, and for genetic interspecies transformation tests. All strains lacked beta-galactosidase; in contrast, beta-xylosidase was always present in the oxidative strains. This test proved to be of value for separating strains able to form acid from carbohydrates (A. anitratum and A. haemolyticus spp haemolyticus) from the non-oxidative strains (A. lwoffi and A. haemolyticus spp alcaligenes). However, the genetic relationship of all strains tested warrants further study before Acinetobacters are grouped into clearly defined species

Sensitivity of Streptococcus pyogenes to sulphamethoxazole, trimethoprim, and cotrimoxazole

When tested on Wellcotest (sensitivity test agar, Wellcome) supplemented with lysed horse blood, 59 clinical isolates of beta-haemolytic streptococci, belonging to several serological groups, are all sensitive to sulphamethoxazole, to trimethoprim, and to cotrimoxazole. The results obtained on diagnostic sensitivity test agar (Oxoid) containing lysed horse blood are comparable. When Mueller Hinton agar (Difco) is used, some of these strains are called resistant: this is especially true in the case of sulphamethoxazole. Furthermore, on Mueller Hinton agar, even supplemented with lysed blood, discrepancies between diameters of inhibition zone and MIC values exist for the three drugs tested, strains with a low MIC being considered resistant by the disc method. Such disagreements are not observed with non-streptococcal control strains