nuclei stained with draq5  (MetaMorph Inc)

Journal: RSC Advances

Article Title: Utilization of metabolic energy in treatment of ocular surface disorders: polyphosphate as an energy source for corneal epithelial cell proliferation

doi: 10.1039/c9ra04409d

Figure Lengend Snippet: Immunofluorescence and ESEM analysis of the outgrown cells, 3 d after initiating of the assays. In the first series the cell layers outgrown from the explants (ex) were stained with (A–C) DRAQ5 to identify the nuclei or double-stained with (D–F) rhodamine/phalloidin and DRAQ5 to express the complex multilayered arrangement of the cells migrated out from the explants; immunofluorescence light microscopy. The rim of the explant is highlighted (ex). In (G–I) higher magnification images of the epithelial cells showing the different degrees of differentiation of the cells; ESEM. In the controls (G) the cells are not continuously attached to each other with cell-to-cell junctions. In addition, a partial exfoliation of the cells is apparent. In contrast, cells migrated from the explants which had been incubated with hp-lysate (HPL) (H), or with polyP (I) smoothly attach to each other and form a homogeneous margin rim of microvilli on their cell surfaces. Surface areas with extensive microvilli decorations are marked (mic).

Article Snippet: Cell outgrowth and differentiation were analyzed after staining with DRAQ5 (nuclei) and rhodamine phalloidin (cytoskeleton), as well as by environmental scanning electron microscopy (ESEM).

Techniques: Immunofluorescence, Staining, Light Microscopy, Incubation

Journal: Physiological Genomics

Article Title: RNO3 QTL regulates vascular structure and arterial stiffness in the spontaneously hypertensive rat

doi: 10.1152/physiolgenomics.00038.2021

Figure Lengend Snippet: MPM imaging of arterial sections. SHR.BN3 ( A ) and SHR ( B ) arterial cross section at ×10 ( i ) and ×40 ( ii ) magnification. Nuclei are labeled with DRAQ5 (blue), elastin (green), and collagen (red). C : average number of nuclei in the adventitia and media of arteries in the SHR.BN3 ( n = 12) and SHR ( n = 17). D : quantification of collagen and collagen bundles in arteries. E : immunohistochemical detection and quantification of Ki-67 in formalin-fixed paraffin-embedded arterial cross sections of SHR.BN3 ( F ; n = 12) and SHR ( G ; n = 17). Slides were stained with DAB and hematoxylin (positive cells brown, nucleus blue). Scale bars equal 100 µm and 50 µm as indicated. Values are expressed as means ± SE. * P < 0.05, statistically significant. DAB, diaminobenzidine; DRAQ5, Deep Red Anthraquinone 5; Ki-67, nuclear protein Ki-67; MPM, multiphoton microscopy; SHR, spontaneously hypertensive rat.

Article Snippet: Using MetaMorph software, nuclei stained with DRAQ5 were counted at three locations within the tunica media and adventitia, and the counts were averaged.

Techniques: Imaging, Labeling, Immunohistochemical staining, Formalin-fixed Paraffin-Embedded, Staining, Microscopy

Journal: Parasitology Research

Article Title: Besnoitia besnoiti– driven endothelial host cell cycle alteration

doi: 10.1007/s00436-020-06744-x

Figure Lengend Snippet: Infection of BUVEC with B. besnoiti tachyzoites and effect of infection on host cell proliferation. Sub-confluent BUVEC were infected with B. besnoiti tachyzoites at an MOI 5:1 and analyzed after 24 hpi. a Quantification of tachyzoites numbers/parasitophorous vacuole during 30 h of infection. The mature structures were observed between 24 and 30 h p.i. After this time point, all parasites were released. b Illustration of non-infected control cells and B. besnoiti– infected BUVEC at 24 h p.i. c Host cell proliferation analyzed by estimating cell numbers of B. besnoiti –infected BUVEC and non-infected controls at 24 h p.i. No statistically significant differences were observed, but it was a hint of decreased number of cells at the B. besnoitia –infected monolayer. d Quantification of binucleated host cells in B. besnoiti –infected BUVEC and non-infected controls. e Illustration of DRAQ5 (vital DNA staining, red)-stained host cell nuclei in B. besnoiti – and T. gondii –infected BUVEC (24 h p.i.) via 3D holotomographic microscopy (note: binucleated phenotype in case of T. gondii –infected BUVEC)

Article Snippet: This was also tested via DRAQ5 (vital staining of nuclei)-based live cell 3D holotomographic microscopy (3D Nanolive®).

Techniques: Infection, Staining, Microscopy