In vitro and in vivo documentation of quantum dots labeled Trypanosoma cruzi--Rhodnius prolixus interaction using confocal microscopy.
Semiconductor quantum dots (QDs) are highly fluorescent nanocrystals markers that allow long photobleaching and do not destroy the parasites. In this paper, we used fluorescent core shell quantum dots to perform studies of live parasite-vector interaction processes without any observable effect on the vitality of parasites. These nanocrystals were synthesized in aqueous medium and physiological pH, which is very important for monitoring live cells activities, and conjugated with molecules such as lectins to label specific carbohydrates involved on the parasite-vector interaction. These QDs were successfully used for the study of in vitro and in vivo interaction of Trypanosoma cruzi and the triatomine Rhodnius prolixus. These QDs allowed us to acquire real time confocal images sequences of live T. cruzi-R. prolixus interactions for an extended period, causing no damage to the cells. By zooming to the region of interest, we have been able to acquire confocal images at the three to four frames per second rate. Our results show that QDs are physiological fluorescent markers capable to label living parasites and insect vector cells. QDs can be functionalized with lectins to specifically mark surface carbohydrates on perimicrovillar membrane of R. prolixus to follow, visualize, and understand interaction between vectors and its parasites in real-time.
Feder
Denise
D
Laboratório de Biologia de Insetos, GBG, Universidade Federal Fluminense (UFF), Niterói, Rio de Janeiro, Brazil. dfeder@vm.uff.br
Gomes
Suzete A O
SA
de Thomaz
André A
AA
Almeida
Diogo B
DB
Faustino
Wagner M
WM
Fontes
Adriana
A
Stahl
Cecília V
CV
Santos-Mallet
Jacenir R
JR
Cesar
Carlos L
CL
D000818:Animals; D002863:Chromogenic Compounds / Q000494:pharmacology*; D006790:Host-Parasite Interactions; D018613:Microscopy, Confocal; D010274:Parasitology / Q000379:methods*; D045663:Quantum Dots; D012239:Rhodnius / Q000469:parasitology*; D013194:Staining and Labeling / Q000379:methods*; D014349:Trypanosoma cruzi / Q000254:growth & development*
D002863:Chromogenic Compounds