Journal: Current Research in Neurobiology

Article Title: Gene delivery to neurons in the auditory brainstem of barn owls using standard recombinant adeno-associated virus vectors

doi: 10.1016/j.crneur.2020.100001

Figure Lengend Snippet: Examples for the expression of serotypes rAAV2/1 and rAAV2/5 in nucleus magnocellularis . A–D: Expression of GFP, indicating rAAV2/1, in NM cell bodies and fibers, three weeks after injection (case #13 in Table 1 ). E–H: Expression of tdTomato, indicating rAAV2/5, in NM cell bodies and fibers, also three weeks after injection (case #14 in Table 1 ). Two examples of labeled cell bodies are highlighted by white arrowheads. Note that in each row (A–C and E–G), the first three panels illustrate identical image frames but separately for 3 fluorescence channels; D and H show the corresponding merged images. Scale bars 50 ​μm. For further information about injection site, volume and expression time, see Table 1 .

Article Snippet: For fluorescent detection and imaging, a Nikon epifluorescence microscope (Nikon Eclipse Ni-E and the associated Nikon imaging software NIS Elements AR.4.30.02 64-Bit), as well as a fluorescence slide scanner (Axio Scan Z1, Zeiss, and the associated software ZEISS Efficient Navigation ZEN 2.6 blue edition) were used.

Techniques: Expressing, Injection, Labeling, Fluorescence

Journal: Current Research in Neurobiology

Article Title: Gene delivery to neurons in the auditory brainstem of barn owls using standard recombinant adeno-associated virus vectors

doi: 10.1016/j.crneur.2020.100001

Figure Lengend Snippet: Examples for the expression of serotypes rAAV2/1 and rAAV2/5 in nucleus laminaris . A–D: Expression of GFP, indicating rAAV2/1, in NM projection fibers ramifying within NL, three weeks after injection (case #13 in Table 1 ). E–H: Expression of tdTomato, indicating rAAV2/5, expressed in NM projection fibers ramifying and terminating within NL, also three weeks after injection (case #14 in Table 1 ). One example each of a labeled fiber (B) and terminals surrounding a (non-fluorescent) cell body (G) are highlighted by gray arrowheads, and also shown in higher-magnification insets. As in Fig. 2 , labeled fibers mostly appear as short fragments because their course was oblique to the sectioning plane. Note also that in each row (A–C and E–G), the first three panels illustrate identical image frames but separately for 3 fluorescence channels; D and H show the corresponding merged images. Scale bars 50 ​μm. For detailed information about injection site, volume and expression time, see Table 1 .

Article Snippet: For fluorescent detection and imaging, a Nikon epifluorescence microscope (Nikon Eclipse Ni-E and the associated Nikon imaging software NIS Elements AR.4.30.02 64-Bit), as well as a fluorescence slide scanner (Axio Scan Z1, Zeiss, and the associated software ZEISS Efficient Navigation ZEN 2.6 blue edition) were used.

Techniques: Expressing, Injection, Labeling, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Enhancement of Intracellular Calcium Ion Mobilization by Moderately but Not Highly Positive Material Surface Charges

doi: 10.3389/fbioe.2020.01016

Figure Lengend Snippet: Dynamic Ca 2+ mobilization in vital fluo-3 loaded MG-63s growing on amino polymers, extracellular matrix (ECM)/peptide motifs and polyelectrolyte multilayers (PEM) in the time frame 0–480 s. The positive control PPAAm (also an amino polymer) and the negative control Ti are embedded for the respective experiment. Shown are time courses of the mean fluorescence intensity of cells (MFI C ) over 8 min: First, the basal Ca 2+ signal is recorded for 180 s. At 180 s the cells are stimulated with ATP (arrowheads) and the Ca 2+ signal is detected (LSM780, Carl Zeiss; ZEN-software). For statistical analyses of the basal MFI, values of the time points 0–170 s are used. For analyses of the MFI after ATP stimulation, values of the time points 190–240 s are used (highlighted in grey). MFI C values are normalized to the mean cell area after 24 h. (Statistics: Kruskal–Wallis and Dunn’s multiple comparison test, p < 0.05; mean ± sem; n ≥ 3 independent approaches, per time point ≥ 30 cells; * indicates significance of modification x to Ti and # significance to PPAAm).

Article Snippet: Oil DIC M27) and the ZEN software (ZEISS efficient navigation, ZEN 2011 SP4, black edition, Carl Zeiss).

Techniques: Positive Control, Polymer, Negative Control, Fluorescence, Software, Comparison, Modification

Journal: International Journal of Molecular Sciences

Article Title: Long-Acting FGF21 Inhibits Retinal Vascular Leakage in In Vivo and In Vitro Models

doi: 10.3390/ijms21041188

Figure Lengend Snippet: Fibroblast growth factor 21 ( FGF21) treatment prevents and FGF21 deficiency Increases vascular endothelial growth factor (VEGF) -induced retinal vascular leakage in the mouse model. ( A ) The time course of this study. ( B – D ) Representative confocal microscopy images of whole-mounted retina. ( E – G ) Images of retina after segmentation with ZEISS Efficient Navigation (ZEN) Intellesis. ( B and E ) Representative retinal image of controls. ( C and F ) Control Phosphate Buffered Saline (PBS) treatment with intravitreal injection of mouse (m) VEGF. ( D and G ) FGF21 (long-acting FGF21; PF05231023) treatment with intravitreal injection of mouse (m) VEGF. The yellow FITC indicates leakage; the red lectin stain indicates vessels and artifacts; the cyan shows background. ( H ) The intensity of retinal vascular leakage was increased in animals injected with mVEGF and treated with PBS (square), compared to controls (circle, n = 9–14; p = 0.046). Leakage intensity was reduced in FGF21 (triangle) versus PBS-treated groups ( n = 14–15; p = 0.01). The data were analyzed by one-way ANOVA with Tukey and were expressed as mean ± standard error (SE) Scale bar is 1 mm. ( I and J ) Representative confocal microscopy images of retinas in Fgf21 +/+ and Fgf21 −/− littermate mice. Representative images are shown. ( K and L ) Images of retina after segmentation with ZEN Intellesis. ( M ) The data were analyzed by unpaired t -test and were expressed as mean ± SE. Fold change compared with the wild type group was calculated ( n = 5–7; p = 0.037). Scale bar is 1mm. i.p., intraperitoneal injection, i.v., intravitreal injection, FGF21, long-acting FGF21 (PF05231023). Fold change was calculated (* p < 0.05).

Article Snippet: The fluorescence intensity of FITC-dextran from vascular leakage was analyzed with the machine learning software “ZEISS Efficient Navigation (ZEN) Intellesis”, to segment and quantify leakage.

Techniques: Confocal Microscopy, Injection, Staining