Measuring Macrophage Phagocytic Efficiency with Primaquine and Photodynamic Therapy

Abstract

Source:
Uju L. Madu¹, Maphori Maliehe¹, Nozethu Mjokane¹, Olihile M. Sebolai<sup>1

1</sup>Department of Microbiology and Biochemistry, University of the Free State.

The video demonstrates evaluating photodynamic treatment's impact on murine macrophage phagocytic efficiency with primaquine. Treated, infected macrophages generate reactive oxygen species, leading to selective fungal death, evident by reduced colonies on the plate.

Protocol

1. Preparation of Cryptococcal cells

1. Streak out the test organism with an inoculation loop onto a sterile yeast-malt-extract (YM) agar plate.
2. Incubate the agar plate at 30°C for 48 h.
3. Use an inoculation loop to scoop 5 single colonies and suspend them in 5 mL of sterile distilled water.
4. Standardize the cells using a McFarland standard to obtain a cell concentration between 0.5 × 10⁵ and 2.5 × 10⁵ colony-forming units (CFU) per mL of RPMI-1640 medium.
5. Dispense a 100 µL cell suspension into wells of microtiter plates that contain seeded macrophages (se....