Dmi6000 b automated microscope

After each treatment condition, the cell morphology was assessed. Leica DMI6000 B Automated Microscope (Wetzlar, Germany) was used to observe and capture MCF-7 cells treated with paclitaxel, propranolol, ICI 118,551, and isoprenaline.

The cellular morphology of SH-SY5Y cell lines was assessed by using Leica DMI6000 B Automated Microscope (Wetzlar, Germany) after all the exposure to the different treatment conditions for 48 h.

Four 5 μm sections from different areas of the brain and eye were stained by periodic acid Schiff hematoxylin (PASH) or hematoxylin and eosin stain respectively. Histopathologic changes (Brain: microglial nodules, perivascular and diffuse inflammation; Retina: disruption of architecture, perivascular and vitreal inflammation) were scored from 0 to 4 similar to previously described criteria^{36 (link),37 (link)}. Sections were also incubated with anti-T. gondii Ab (BioGenex) and Tomato lectin-DyLight 488 (Vector laboratories) that efficiently labels neural endothelial cells^{38 (link)} (at 0.5 μg/ml it stains neural endothelial cells rather than microglia) or anti-CD31 Ab (Elabscience). Coronal sections at the septo-diencephalic region (level of the thalamus) were examined at X400. The numbers of clusters of T. gondii parasites within tomato lectin⁺ elongated structures (endothelial cells) were counted per whole coronal section. Brain sections were also stained with anti-LC3 (Abgent) or anti-LAMP-1 (Developmental Studies Hybridoma Bank) Abs. Accumulation of LC3 or LAMP-1 around T. gondii located in endothelial cells was defined as the presence of a ring-like structure that surrounds the parasite^{21 (link),22 (link)}. Slides were analyzed using Leica DMI 6000 B automated microscope equipped for epifluorescence microscopy.

The cellular morphology of SH-SY5Y and HT-22 cell lines was assessed by using Leica DMI6000 B Automated Microscope (Wetzlar, Germany) after all the treatment conditions before the cell viability assays.

Mammalian cells challenged with RFP T. gondii were fixed with 4% paraformaldehyde at 5 h and stained with rabbit anti-LC3 antibody (MBL International, Woburn, MA) plus Alexa 488-conjugated secondary antibody (Invitrogen, Carlsbad, CA). Mammalian cells infected with YFP T. gondii were fixed at 8 h and stained with mouse anti-human LAMP-1 mAb (Developmental Studies Hybridoma Bank; Iowa City, IA) plus Alexa 555-conjugated secondary antibody (Jackson ImmunoResearch Laboratories Inc.). Accumulation of LC3 or LAMP-1 around T. gondii was defined as the presence of a ring-like structure that surrounds the parasite [62 (link), 63 (link)]. At least 50 cells per well (duplicate or triplicate wells per group per experiment) were counted manually. CHO cells were challenged with YFP T. gondii and fixed at 3 min. Monolayers were stained with mouse anti-phospho-Y397 FAK antibody (BD Biosciences) plus Alexa 555-conjugated secondary antibody (Jackson ImmunoResearch Laboratories Inc.) and rabbit anti-RON4 antibody (gift from John Boothroyd) followed by incubation with goat anti-rabbit Alexa 647-conjugated secondary antibody (Jackson ImmunoResearch Laboratories Inc.). Specificity of staining was determined by incubating monolayers with secondary antibody alone. Slides were analyzed using Leica DMI 6000 B automated microscope equipped for epifluorescence microscopy.

Cell morphologies were assessed using the Leica DMI6000 B Automated Microscope (Wetzlar, Germany) to observe and capture images of SH-SY5Y cells after all the treatment conditions.

Cell morphology was assessed using a Leica DMI6000 B automated microscope (Wetzlar, Germany). After all treatments, cell morphology was examined and images were captured using Leica LAS X imaging software v3.7.4 (Leica Microsystems, Wetzlar, Germany).