Respected Sir,I am Sakshi Jain, a Ph.D. student from the Indian...

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sakshi jain
May 31, 2021
protocol Protocol: Isolation and Infection of Drosophila Primary Hemocytes
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 sur
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Charles Tracy Author Answered Jun 2, 2021

UT Southwestern Medical Center Dallas

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.
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