I am interested to track bacterial engulfment into phagosome, phaoglysosomal fusion and measure lysosomal pH. That's why, need marker for phagosome, lysosome and a way to quantify lysosomal pH.
1) Can anyone suggest phagosomal marker for macrophages?
2) To quantify lysosomal pH, I am not sure whether to choose Lysotracker or Lysosensor? Any suggestion?
Thanks
1) Rab5a, Rab7 and LAMP-2 are good early to late phagosome maturation markers.
2) The advantages of using Lysosensors are, they have higher quantum yields at lower pH ranges to allow visualization of the lysosomes. They are also relatively less toxic compared to LysoTracker probes. The disadvantages are, they need relatively longer internalization time and are less brighter as it was said that LysoSensor exhibit a pH-dependent increase in fluorescence intensity upon acidification, in contrast to the LysoTracker.
Hi Abdullah
As Vinod Nadella mentioned Rab7, Lamp1 and Lamp2 are good markers. However if you are planning to go live, I would recommend the following.
1. If you have a bacteria expressing GFP and mCherry together, they can provide a rough readout of phagolysosomal processing as the GFP signal will slowly die out as the endosomal pH drops below 5, while mCherry will still be present. The degradation of mCherry will lead to total signal loss indicating processing by lysosomal hydrolases.
2. Another tool to monitor pH in phagosomal maturation are the pH rhodo indicators (link below). They also have bacteris conjugated to these probes, if it works for you.
https://www.thermofisher.com/pt/en/home/brands/molecular-probes/key-molecular-probes-products/phrodo-indicators.html
https://www.thermofisher.com/pt/en/home/brands/molecular-probes/key-molecular-probes-products/phrodo-indicators.html#phagocytosis
Hope this helps. Good luck
Best
Aparajita
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