Journal: Scientific Reports

Article Title: SAS6-like protein in Plasmodium indicates that conoid-associated apical complex proteins persist in invasive stages within the mosquito vector

doi: 10.1038/srep28604

Figure Lengend Snippet: T. gondii expressing endogenous SAS6L with C-terminal fusion to HA-APEX fixed in host vacuoles and stained for HA (green) and either IMC1 (red, a and c ) or centrin (red, b ) ( a ) 3D-SIM super-resolution images with cell apices facing the viewer showing SAS6L rings (arrows, insets) and apices in profile showing SAS6L flush with the apical IMC. ( b ) Widefield images of parasites at different points of the cell cycle indicated by centrosome number and position (stained for centrin): interphase with single centrosome per cell (i); newly divided centrosomes close together (ii, upper vacuole); divided centrosomes migrated further apart (ii, lower vacuole; and iii); cells with nascent apical complexes (iv, arrowheads). ( c ) Widefield images of parasites showing daughter pellicle formation (stained for IMC1): before daughter pellicle formation (i and ii, upper vacuoles); small pellicle cups (i and ii, lower vacuoles; iii, upper vacuole); mid pellicle development (iii, lower vacuole). Scale bars = ( a ) 5 μm and 500 nm (inset); and ( b , c ) 10 μm.

Article Snippet: P. bergehi images were captured as described for live imaging using a 100x oil immersion objective, and T. gondii images were captured using a Nikon Ti-E widefield inverted microscope with a Hamamatsu Orca-Flash4.0 CMOS camera.

Techniques: Expressing, Staining

Journal: Scientific Reports

Article Title: Remote-refocusing light-sheet fluorescence microscopy enables 3D imaging of electromechanical coupling of hiPSC-derived and adult cardiomyocytes in co-culture

doi: 10.1038/s41598-023-29419-w

Figure Lengend Snippet: Synchronized contraction between spatially separated adult-CM in day 0, 1, 2 co-culture imaged under widefield transillumination. ( a–c ) Top images show two merged frames: at peak of contraction (cyan), and at baseline (red), taken t = 2.5 s apart for samples on ( a ) day 0, ( b ) day 1 and ( c ) day 2 of co-culture. Orange arrows indicate some of the adult cardiomyocytes on top of the layer of hiPSC CM co-culture. ( a–c ) Bottom row shows zoomed in ROI indicated by the yellow rectangles in the top row, with green arrows pointing out some of the contraction movement between the merged frames (see Supplementary Video for 50 s timelapse). ( d–f ) Images for days 0, 1, and 2, calculated as the standard deviation across the whole acquisition, indicating movement during acquisition. The yellow rectangles indicate the zoomed in ROI shown in the bottom row of ( a–c ). Scalebar: 100 µm.

Article Snippet: For imaging over a larger FOV than achievable with the LSFM system magnification, complementary brightfield transillumination and epi-fluorescence 2D-timelapse of the hiPSC-CM and adult-CM co-culture was carried out on an inverted widefield microscope (IX73, Olympus) equipped with LED illumination (CoolLED pE300) and a B/W CCD digital camera (Hamamatsu C4742-95-12ER).

Techniques: Co-Culture Assay, Standard Deviation

Journal: Scientific Reports

Article Title: Remote-refocusing light-sheet fluorescence microscopy enables 3D imaging of electromechanical coupling of hiPSC-derived and adult cardiomyocytes in co-culture

doi: 10.1038/s41598-023-29419-w

Figure Lengend Snippet: Widefield fluorescence imaging of calcium transients in adult-CM and hiPSC-CM co-culture on day 0 (left), day 1 (middle) and day 2 (right). Single frames at baseline ( a–c ) and peak of first transient ( d–f ), with frames from the same sample displayed on the same intensity scale. ( h–j ) Standard deviation calculated across the whole acquisition. Orange arrows indicate examples of coupled adult-CM. ( k–m ) Fluorescence intensity variation through hiPSC-CM (blue square ROI in middle row) and adult-CM (orange square ROI in middle row) indicating synchronized calcium transients. Scalebar: 100 µm. The corresponding timelapse is shown in Supplementary Video .

Article Snippet: For imaging over a larger FOV than achievable with the LSFM system magnification, complementary brightfield transillumination and epi-fluorescence 2D-timelapse of the hiPSC-CM and adult-CM co-culture was carried out on an inverted widefield microscope (IX73, Olympus) equipped with LED illumination (CoolLED pE300) and a B/W CCD digital camera (Hamamatsu C4742-95-12ER).

Techniques: Fluorescence, Imaging, Co-Culture Assay, Standard Deviation