Tcs sp2 abos confocal laser scanning microscope

Leishmania donovani promastigotes (strain MHOM/SD/00/1S-2D) were resuspended in Hanks' medium supplemented with 10 mM D-glucose (HBSS-Glc) at 2 × 10⁷ cells/mL. Afterwards, 9 μM Bodi Fluor 488-labeled PMM-Tat conjugate 6 was added to the parasite suspension and incubated for 4 h in HBSS-Glc. After incubation, non-incorporated 6 was removed by washing the cells with HBSS-Glc. Finally, cells were labeled with DAPI (5 μg/mL, 20 min) before their observation, unfixed, in a Leica TCS–SP2 ABOS confocal laser scanning microscope.

L. donovani promastigotes at 2 ×
10⁷ cells/mL were incubated
with ECP-Alexa at 4 μM for 4 or 12 h in Hank′s buffer
or complete media, respectively. For infected macrophages, only the
second condition was used. After incubation, the cells were washed
with PBS and labeled with DAPI (5 μg/mL) 10 min before observation
in a Leica TCS-SP2 ABOS confocal laser scanning microscope, without
fixation. Colocalization experiments were carried out by incubation
of promastigotes with 10 μM ECP-H15A Alexa plus 10 mg/mL rhodamine
B isothiocyanate dextran (∼70 kDa) for 6 h. Pearson correlation
coefficient was obtained with ImageJ software (Version 1.53r).