observer z1 spinning disc microscope  (Carl Zeiss)

Journal: bioRxiv

Article Title: The SV40 T-ag nuclear localization signal affects Toxoplasma gondii viability by targeting importin α

doi: 10.1101/2025.07.20.665720

Figure Lengend Snippet: (a) T. gondii tachyzoite vacuoles expressing 1xGFP construct alone and when fused with SV40-NLS. The images were captured on Zeiss observer Z1 Spinning Disc microscope with 40X objective (scale bar 50 μm); (b) Percentage of GFP-positive vacuoles per field were counted (N = 3) and the mean ± SEM was plotted; (c) The alanine mutations (bold) of the SV40-NLS motif generated in our study are shown, with their dissociation constant (K d ) values and the cNLS score; (d) Percentage of GFP-positive vacuoles per field were counted (N = 3) with the mean ± SEM plotted; (e) Tachyzoite vacuoles expressing 2xGFP construct alone and when fused with SV40-NLS (scale bar 50 μm); (f) Percentage of GFP-positive vacuoles per field were counted (N = 3) and the mean ± SEM was plotted. A significant difference of p < 0.01 and p < 0.0001 is indicated by * and ****, respectively, on performing the Mann-Whitney test for all plotted graphs.

Article Snippet: To quantify GFP-positive parasitophorous vacuoles, 15 fields were imaged from each replicate in the Zeiss Observer Z1 Spinning Disc microscope at 40X.

Techniques: Expressing, Construct, Microscopy, Generated, MANN-WHITNEY

Journal: bioRxiv

Article Title: The SV40 T-ag nuclear localization signal affects Toxoplasma gondii viability by targeting importin α

doi: 10.1101/2025.07.20.665720

Figure Lengend Snippet: (a) WT-TgImpα-HA, AAA-TgImpα-HA, and KRR-TgImpα-HA localize in the nucleus and cytoplasm of T. gondii tachyzoites. Images were captured in a Zeiss LSM 780 Confocal microscope with 100X objective (scale bar 5 μm); (b) Transient co-transfection of WT and variants of TgImpα-HA (red) with SV40-NLS-2xGFP (green) resulted in live SV40-NLS-GFP expressing vacuoles. Images were captured in Zeiss Observer Z1 Spinning Disc microscope with 40X objective (scale bar 30 μm); (c) Graph shows mean ± SEM of the percentage of vacuoles after transient co-transfection expressing only SV40-NLS-2xGFP or 2xGFP (green), only TgImpα-HA (red) or both (yellow) per DAPI counts (N = 3). Mann-Whitney test performed shows a significant difference of p < 0.003 (***) or p < 0.0001 (****) for the yellow bar in the plotted graph; (d) Graph shows mean ± SEM of the percentage of vacuoles in the stable lines of TgImpα-HA (wild-type and variants) transfected with SV40-NLS-2xGFP or 2xGFP (N = 2); (e) Graph shows mean ± SEM of the CTCF intensity estimated of TgImpα-HA in transiently transfected and stable lines for 30-40 vacuoles. The Mann-Whitney test performed shows a significant difference of p < 0.0001.

Article Snippet: To quantify GFP-positive parasitophorous vacuoles, 15 fields were imaged from each replicate in the Zeiss Observer Z1 Spinning Disc microscope at 40X.

Techniques: Microscopy, Cotransfection, Expressing, MANN-WHITNEY, Transfection

Journal: bioRxiv

Article Title: The SV40 T-ag nuclear localization signal affects Toxoplasma gondii viability by targeting importin α

doi: 10.1101/2025.07.20.665720

Figure Lengend Snippet: (a) GFP-His and SV40-NLS-GFP-His recombinant proteins (14 μg/ml) were transfected into HFF cells and incubated for 24 hours. The images were captured on Zeiss observer Z1 Spinning Disc microscope with 63X objective (scale bar 30 μm); (b) Graph shows mean ± SEM of the CTCF intensity estimated of GFP in 30 fields at varying concentrations of proteins (N = 2); (c) Cell viability MTT assay performed on HFF when treated with the proteins in the respective concentration range (N = 3).

Article Snippet: To quantify GFP-positive parasitophorous vacuoles, 15 fields were imaged from each replicate in the Zeiss Observer Z1 Spinning Disc microscope at 40X.

Techniques: Recombinant, Transfection, Incubation, Microscopy, MTT Assay, Concentration Assay

Journal: Stem Cells

Article Title: Impact of pro-inflammatory cytokine preconditioning on metabolism and extracellular vesicles in feline mesenchymal stromal cells: a preliminary study

doi: 10.1093/stmcls/sxaf014

Figure Lengend Snippet: Impact of TNF-α/IFN-γ treatment on FeASCs’ mitochondrial metabolism. (A) Oxygen consumption kinetics analysis as an indicator of basal respiration and ATP production rates. (B) Glycolytic capacity and glycolytic reserves determined in native and preconditioned ASCs. (C) representative mitochondrial depolarization Plot (D) Assessment of mitochondrial transmembrane potential. (E) MitoRed-stained mitochondria with DAPI counterstained nuclei were observed under a confocal scanning laser microscope. (F) Morphometric analysis of mitochondria subtypes using MicroP program projection. (G) RNA sequencing of mitochondrial-related gene network associated with biogenesis and metabolism. Representative data is shown as mean ± SD. n ASCs, native FeASCs; p ASCs, cytokine-primed FeASCs; n EVs, EVs isolated from native FeASCs; p EVs, EVs isolated from cytokine-primed FeASCs. An asterisk (*) indicates a comparison among AD-MSCs and PD-MSCs. P < .05, ** P < .01, **** P < .0001.

Article Snippet: The fluorescence signals were observed and imaged using a confocal microscope (Observer Z1 Confocal Spinning Disc V.2 Zeiss with live imaging chamber) at 400× magnification.

Techniques: Staining, Microscopy, RNA Sequencing, Isolation, Comparison