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  • ASAS Flow Cytometry Presentation - Theo Online

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    Pailago Kate Jan Queen
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    This document discusses using flow cytometry to study the interactions between Trichomonas vaginalis and Prevotella bivia. Specifically, it aims to use flow cytometry to generate growth curves of the two microbes, study their growth effects when exposed to each other's conditioned media, and examine T. vaginalis's ability to adhere to and degrade mucus. The document outlines how flow cytometry provides faster, more accurate and automated cell counts compared to traditional methods, and discusses how it was successfully used to create growth curves of T. vaginalis and P. bivia over 24 hours. In conclusion, the benefits of flow cytometry for microbiology applications are highlighted, including time savings, accuracy, automation, and ability to differentiate

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    ASAS Flow Cytometry Presentation - Theo online (1)

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    This document discusses using flow cytometry to study the interactions between Trichomonas vaginalis and Prevotella bivia. Specifically, it aims to use flow cytometry to generate growth curves of the two microbes, study their growth effects when exposed to each other's conditioned media, and examine T. vaginalis's ability to adhere to and degrade mucus. The document outlines how flow cytometry provides faster, more accurate and automated cell counts compared to traditional methods, and discusses how it was successfully used to create growth curves of T. vaginalis and P. bivia over 24 hours. In conclusion, the benefits of flow cytometry for microbiology applications are highlighted, including time savings, accuracy, automation, and ability to differentiate

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    Download as PPTX, PDF, TXT or read online from Scribd
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    48 views15 pages

    ASAS Flow Cytometry Presentation - Theo Online

    Uploaded by

    Pailago Kate Jan Queen
    This document discusses using flow cytometry to study the interactions between Trichomonas vaginalis and Prevotella bivia. Specifically, it aims to use flow cytometry to generate growth curves of the two microbes, study their growth effects when exposed to each other's conditioned media, and examine T. vaginalis's ability to adhere to and degrade mucus. The document outlines how flow cytometry provides faster, more accurate and automated cell counts compared to traditional methods, and discusses how it was successfully used to create growth curves of T. vaginalis and P. bivia over 24 hours. In conclusion, the benefits of flow cytometry for microbiology applications are highlighted, including time savings, accuracy, automation, and ability to differentiate

    Copyright:

    © All Rights Reserved
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    of 15

    Integration of Flow

    Cytometry in
    Microbiology
    Theodore D’Souza
    Background

     Molecular microbiology
     Study the interactions between Trichomonas vaginalis & Prevotella bivia
     T. vaginalis
     Causes Trichomoniasis
     ~160 million cases per year worldwide1
     Most common non-viral STI in the world
     P. bivia
     Anaerobic bacteria involved in Bacterial Vaginosis
     Most common vaginal infection in females aged between 15-44 2

    1. World Health Organisation. (2011). Prevalence and incidence of selected sexually transmitted infections. Switzerland. WHO.

    2. Centers of Disease Control and Prevention. (2014). Bacterial Vaginosis – CDC Fact Sheet. United States of America. CDC.
    Background

     Low usage in microbiology


    BD Calibur & BD Accuri
    C Sampler
    Flow cytometry use in my project:
    Specific Aims
     Growth curves of T. vaginalis & P. bivia
     Growth effects of conditioned media
     Adhesion + degradation to mucus by T. vaginalis
     All aims need real cell counts
    Flow cytometry use in my project:
    Flow vs Traditional Alternatives
     Growth curves of T. vaginalis & P. bivia
     OLD:
     Before experiment: Count using Haemocytometer or Spectrophotometer
     After experiment: Need media plates takes 3-4 days for T. vaginalis

    takes 24 hours for P. bivia


    NEW: All done using FACS
     Growth effects of conditioned media
     OLD: Same as above
     Adhesion + degradation by T. vaginalis
     OLD: Count under a microscope at each time point
    Flow cytometry use in my project:
    Use
     Stain cells
     Complete experiment
     Load into plate
     Set parameters
     Run
     Go have a coffee 
    Flow cytometry use in my project:
    Outcomes
     Successfully created growth curves for T. vaginalis & P. bivia
    Flow cytometry use in my project:
    Outcomes
    Flow cytometry use in my project:
    Outcomes
    Flow cytometry use in my project:
    Outcomes

    G3 cells/50µl over a 24 hour period


    20000

    18000

    16000

    14000
    Cells (per 50ul)

    12000

    10000

    8000

    6000

    4000

    2000

    0
    0 3 6 8 10 12 14 16 24

    Time (hours)
    Flow cytometry use in my project:
    Outcomes
    Benefits: General & Specific

     Time
     Accuracy/Measure viability
     Automation
     Differentiation between different cell populations
     User friendly
    Acknowledgements

     Dr. Augusto Barbosa


     Dr. Anna Brooks
     Alicia Didsbury
     Barbosa lab group

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