Evaluation of Flow Cytometry for Cell Count and Detection of Bacteria in Biological Fluids

Dossou, Nefert Candace; Gaubert, I; Moriceau, C.; Cornet, Édouard; Hello, Simon Le; Malandain, D.

doi:10.1128/spectrum.01830-21

Cited by 7 publications

(2 citation statements)

References 13 publications

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“…First, cells are tagged with fluorescence barcodes and are developed into a single-cell flow via hydrodynamic focusing; next, fluorescent dyes on the same cell are simultaneously activated by laser and their fluorescent spectra are detected simultaneously by multiple channels; fluorescence-labeled cells are distinguished by their fluorescent signals; depending on their fluorescent signals, cells are divided into different groups and sorted into different collection tubes via an electric field controlled by a computer system. FACS is a powerful technology to quickly distinguish specific cells from a large pool and accurately analyze their characteristics. − FACS has a high sorting speed, which reaches up to thousands of cells per second passing one by one in a flow cytometer . However, the main disadvantage of FACS is the overlapping of fluorescent signals, making it difficult to distinguish different fluorescent dyes and greatly limiting the number of fluorescent dyes available for fluorescent barcodes …”

Section: Fluorescence Barcodementioning

confidence: 99%

Digital Barcodes for High-Throughput Screening

Yang,

Chen,

Xiao

et al. 2024

Chem Bio Eng.

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High-throughput screening is an indispensable technology in drug discovery, cancer therapy, and disease diagnosis, and it could greatly reduce time cost, reagent consumption, and labor expense. Here, four high-throughput screening methods with high sensitivity and accessibility are discussed in detail. Fluorescence, DNA, heavy metal, and nonmetal isotope barcodes, which generally label antibodies, proteins, and saccharides to identify cells, are detected by flow cytometry, second-generation DNA sequencing, mass cytometry, and second-ion mass spectrometry, respectively. Encoding binary information in barcodes, labeling individual cells by barcodes, performing the characterization of cells together, and identifying the result belonging to individual cells via barcodes are the main steps for high-throughput screening. Applications of the four digital barcodes in high-throughput screening for both in vitro and in vivo tests are described in detail, and their advantages and disadvantages are also summarized. High-throughput screening has provided a powerful platform widely accessible for multidisciplinary studies and has greatly sped up the progress of drug discovery, disease diagnosis, and cancer therapy.

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Section: Fluorescence Barcodementioning

confidence: 99%

Digital Barcodes for High-Throughput Screening

Yang,

Chen,

Xiao

et al. 2024

Chem Bio Eng.

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“…Each material was tested with each biofilm dislodging Serial dilution, plating and colony enumeration assumes that the biofilm is truly dispersed into individual cells. Flow cytometry (Sysmex ® ) [61] was used to characterize the size distribution of material to confirm the single cell dispersal.…”

Section: Biofilm Removal Treatmentmentioning

confidence: 99%

What is the best technic to dislodge Staphylococcus epidermidis biofilm on medical implants?

et al. 2022

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Background Bacterial biofilm can occur on all medical implanted devices and lead to infection and/or dysfunction of the device. In this study, artificial biofilm was formed on four different medical implants (silicone, piccline, peripheral venous catheter and endotracheal tube) of interest for our daily clinical and/or research practice. We investigated the best conventional technic to dislodge the biofilm on the implants and quantified the number of bacteria. Staphylococcus epidermidis previously isolated from a breast implant capsular contracture on a patient in the university hospital of Dijon was selected for its ability to produce biofilm on the implants. Different technics (sonication, Digest-EUR®, mechanized bead mill, combination of sonication plus Digest-EUR®) were tested and compared to detach the biofilm before quantifying viable bacteria by colony counting. Results For all treatments, the optical and scanning electron microscope images showed substantial less biofilm biomass remaining on the silicone implant compared to non-treated implant. This study demonstrated that the US procedure was statistically superior to the other physical treatment: beads, Digest-EUR® alone and Digest-EUR® + US (p < 0.001) for the flexible materials (picc-line, PIV, and silicone). The number of bacteria released by the US is significantly higher with a difference of 1 log on each material. The result for a rigid endotracheal tube were different with superiority for the chemical treatment dithiothreitol: Digest-EUR®. Surprisingly the combination of the US plus Digest-EUR® treatment was consistently inferior for the four materials. Conclusions Depending on the materials used, the biofilm dislodging technique must be adapted. The US procedure was the best technic to dislodge S. epidermidis biofilm on silicone, piccline, peripheral venous catheter but not endotracheal tube. This suggested that scientists should compare themselves different methods before designing a protocol of biofilm study on a given material.

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