Plasmodium berghei - infected red cells sorted according to DNA content

Area of Science:

• Parasitology
• Cell Biology
• Biochemistry

Background:

• Plasmodium berghei infects red blood cells, causing malaria.
• Accurate separation of infected cells is crucial for research.

Purpose of the Study:

• To develop a DNA-content-based cell-sorting method for Plasmodium berghei-infected red blood cells.
• To enable biochemical and immunochemical studies of infected cells.

Main Methods:

• Selective uptake of 33258 Hoechst, a DNA-binding dye, by infected red blood cells.
• Incubation at 37°C for dye uptake, followed by washing at 4°C.
• Cell sorting based on fluorescence intensity proportional to parasite DNA content.

Main Results:

• Uninfected cells are non-fluorescent; infected cells exhibit fluorescence correlating with parasite DNA.
• Sorted infected cells, incubated in nutrient medium, caused lethal P. berghei infections in mice.
• The method allows collection of viable infected red blood cells for further analysis.

Conclusions:

• The 33258 Hoechst dye-based cell-sorting technique effectively isolates viable Plasmodium berghei-infected red blood cells.
• This method facilitates biochemical and immunochemical investigations of malaria parasites.
• The technique is simple and applicable to cells at various infection stages.