Charles Tracy —BIO-PROTOCOL

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Helmut Krämer Department of Neuroscience, University of Texas Southwestern Medical Center, USA
2 protocols

Charles Tracy

Department of Neuroscience, University of Texas Southwestern Medical Center, USA

Research focus

Immunology

2 Author merit
Reviewer merit
Editor merit

2 Protocols published

Isolation and Infection of Drosophila Primary Hemocytes

Authors: Charles Tracy and Helmut Krämer, date: 06/05/2017, view: 8178, Q&A: 1

Phagocytosis of invading pathogens and their subsequent clearance in lysosomes is important for organismal fitness. We have devised the following protocol to extract phagocytic hemocytes from wild-type and mutant Drosophila larvae and ...

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sakshi jain

Indian Institute of Science

Respected Sir,
I am Sakshi Jain, a Ph.D. student from the Indian Institute of Science, Bangalore. I have a doubt regarding the quantification of the number of bacteria engulfed per cell. In Image J or Fiji, how are you accounting that the bacteria is inside the cell and not on the surface?

Thanking You in advance.

5/31/2021 4:02:19 AM Reply

Charles Tracy

Department of Neuroscience, University of Texas Southwestern Medical Center, USA

After an initial pulse to allow bacteria to bind and chase of the desired time, cells are fixed and cellular actin stained with phalloidin, which allows for visualization of the cell membranes by confocal microscopy. To be counted as intracellular, a bacterium must be fully engulfed within the phalloidin-stained membrane. Commonly, DIC images are also taken while imaging to further assist in this determination.

2021-06-02 08:57:35

Escherichia coli Infection of Drosophila

Authors: Charles Tracy and Helmut Krämer, date: 05/05/2017, view: 6943, Q&A: 0

Following septic insults, healthy insects, just like vertebrates, mount a complex immune response to contain and destroy pathogens. The failure to efficiently clear bacterial infections in immuno-compromised fly mutants leads to higher mortality ...

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