zeiss axio epifluorescence microscope  (Carl Zeiss)

Journal: bioRxiv

Article Title: Morphological and molecular effects of short-term water deficiency in barley stamen maturation

doi: 10.1101/2024.11.19.624362

Figure Lengend Snippet: A ) DAPI staining of pollen nuclei. vN, vegetative nucleus. sN, sperm cell nuclei. Bars, 10 µm. B ) Pollen viability in control or drought-affected stamens (confirmed failure of starch accumulation) assayed simultaneously with fluorescein diacetate (FDA) and propidium iodide (PI) dyes. FDA fluoresces (green) in viable cells. Arrowheads indicate pollen with only background fluorescent signal, and thus considered non-viable or dead. Instead, PI accumulates inside dead cells and increases the intensity of fluorescence (red, arrowheads). Note that all pollen grains have a relatively high red signal, a combination of both autofluorescence and emission from PI accumulated on the surface of viable cells (images obtained with a normal epifluorescence microscope). Bars, 200µm. C ) Quantification of pollen viability determined by FDA-PI staining. Each data point is the ratio of viable pollen counted from three different florets in one inflorescence. Significance of difference between control and drought was determined with a one-tailed t-test. n = 4 inflorescences (598 and 464 total pollen grains scored for control and drought, respectively).

Article Snippet: For visualizing microspores within anthers at stages W7.5 – W8.25, anthers were mounted in perfluoroperhydrophenanthren (Sigma-Aldrich/Merck) and imaged with a Zeiss Axio Scope.A1 light (epifluorescence) microscope combined with an Axiocam 512 color camera.

Techniques: Staining, Control, Starch, Fluorescence, Microscopy, One-tailed Test

Journal: Cells

Article Title: Piezo1 Is Required for Myoblast Migration and Involves Polarized Clustering in Association with Cholesterol and GM1 Ganglioside

doi: 10.3390/cells12242784

Figure Lengend Snippet: The increased Fura2 signal at the cell front upon Piezo1 activation by Yoda1 is abrogated by Piezo1 silencing. Cells transfected with a negative control siRNA (siCTL, white) or with two different siRNAs targeting Piezo1 (siPiezo1 #1 and #2, green) were left untreated or stimulated with 0.5 µM Yoda1 (black) for 90 s under resting conditions ( A ), or for 30 min and allowed to migrate for 5 h ( B – G ), then incubated with Fura2-AM and imaged at 340 and 380 nm with an epifluorescence microscope. The data obtained with siPiezo1 #1 and #2 were then pooled (siPiezo1-all). ( A ) The proportion of cells showing a Ca 2+ response after 90 s of stimulation with 0.5 µM Yoda1. Significant decrease upon Piezo1 silencing observed, according to the Fisher test. ( B ) Global Fura2-AM ratio (340/380) in whole migrating C2C12 cells ( n = 4 independent experiments). ( C ) Heatmap of the ratio (340/380) along the main axis of each individual migrating cell. ( D – F ) Mean ratio (340/380) along the main cell axis from rear to front in siCTL- ( D ), siPiezo1 #1- ( E ), or siPiezo1 #2-transfected cells ( F ). ( G ) Violin plot of the ratio of the front to the center part of each cell ( n = 250 cells from 4 independent experiments). Dotted line indicate the no polarization value; Straight lines in the plots indicate medians, while dotted lines in the plots indicate the interquartile range, Evaluated by Kruskal–Wallis test, followed by Dunn’s multiple comparisons test. The statistics above the columns refer to the corresponding control while the statistics between different groups are indicated with bars on top of graphs (*, p -value < 0.05).

Article Snippet: The Fura2-AM-loaded cells were placed on the stage of an upward epifluorescence microscope (Zeiss Axio Examiner, Oberkochen, Germany) and continuously superfused using Krebs solution.

Techniques: Activation Assay, Transfection, Negative Control, Incubation, Microscopy