Gislaine Vieira-Damiani, Marna Elise Ericson, Marilene Neves da Silva, Kalpna Gupta, Tania Benetti Soares, Amanda Roberta de Almeida, Vitor Bianchin Pelegati, Mariana Ozello Baratti, Carlos Lenz Cesar, Maria Letícia Cintra and Paulo Eduardo Neves Ferreira Velho
Bartonella henselae is a causative agent of anemia, cat scratch disease, bacillary angiomatosis, recurrent fever, hepatitis, endocarditis, chronic lymphadenopathy, joint and neurological disorders. B. henselae are intra-erythrocytic bacteria. The goal of this study was to visualize the B. henselae invasion into enucleated human red blood cells in real time using bacterium endogenous fluorescence. We took advantage of the unique fluorescence emission spectral profile of the bacteria. We used a linear unmixing approach to separate the fluorescence emission spectra of human erythrocytes from native B. henselae when excited at 488nm. Human blood samples were inoculated with B. henselae and incubated for 60 hours. 3-D live images were captured at select intervals using multi-photon laser scanning microscopy. Uninfected blood samples were also analyzed. This study revealed bacteria entering mature erythrocytes over a 60 hour time period.