Journal: BMC Microbiology

Article Title: Localization of Staphylococcus aureus in tissue from the nasal vestibule in healthy carriers

doi: 10.1186/s12866-017-0997-3

Figure Lengend Snippet: Localization of S. aureus in the upper part of epidermis. Inset represents a zoomed portion of the image. Scale bar is in micrometers. We used a confocal laser scanning microscopy (CLSM) ×63 objective. a-b S. aureus is localized on the border between stratum corneum and stratum granulosum. Green fluorescence labeling of S. aureus , blue fluorescence labeling of keratinocyte cell nuclei and S. aureus DNA. Primary antibody rabbit polyclonal antibody to S. aureus (ab20920, Abcam), secondary antibody Alexa Fluor 488® goat anti-rabbit IgG (Molecular Probe™, Thermo Fisher Scientific), and DRAQ5 (BioStatus). c S. aureus in stratum corneum. The same labeling is used as in a-b, but in addition, we used Alexa Fluor 594 Phalloidin (A12381, Molecular Probes™, Thermo Fisher Scientific) staining actin red. d Combined CLSM and light microscopy of nasal tissue section. S. aureus seen as red cocci in pairs, presumably on the border between stratum corneum and stratum granulosum. Primary S. aureus rabbit polyclonal antibody (ab20920, Abcam) and a secondary antibody Alexa Fluor 546 goat anti-rabbit IgG (Molecular Probes™, Thermo Fisher Scientific) were used

Article Snippet: S. aureus is labeled with primary rabbit polyclonal antibody to S. aureus (Abcam), secondary antibody Alexa Fluor 488 goat anti-rabbit IgG (green) (Molecular ProbesTM, Thermo Fisher Scientific), DRAQ5 (BioStatus) for keratinocyte nuclei (blue) and Alexa Fluor 594 Phalloidin (A12381, Molecular ProbesTM; Thermo Fisher Scientific) for actin (red).

Techniques: Confocal Laser Scanning Microscopy, Fluorescence, Labeling, Staining, Light Microscopy

Journal: BMC Microbiology

Article Title: Localization of Staphylococcus aureus in tissue from the nasal vestibule in healthy carriers

doi: 10.1186/s12866-017-0997-3

Figure Lengend Snippet: Localization of S. aureus in stratum spinosum by CLSM. Inset represents a zoomed portion of the image. Scale bar is in micrometers. We used a CLSM ×63 objective. The sections are oriented so that the outermost epidermis layer is shown in the upper left corner. a-b Green fluorescence labeling of S. aureus , blue fluorescence labeling of keratinocyte cell nuclei and S. aureus DNA. c Same labeling as in a and b, but in addition we used red fluorescence labeling of actin. Primary antibody rabbit polyclonal antibody to S. aureus (ab20920, Abcam), secondary antibody Alexa Fluor 488® goat anti-rabbit IgG (Molecular Probe™, Thermo Fisher Scientific), Alexa Fluor 594 Phalloidin (Molecular Probes™, Thermo Fisher Scientific) and DRAQ5 (BioStatus)

Article Snippet: S. aureus is labeled with primary rabbit polyclonal antibody to S. aureus (Abcam), secondary antibody Alexa Fluor 488 goat anti-rabbit IgG (green) (Molecular ProbesTM, Thermo Fisher Scientific), DRAQ5 (BioStatus) for keratinocyte nuclei (blue) and Alexa Fluor 594 Phalloidin (A12381, Molecular ProbesTM; Thermo Fisher Scientific) for actin (red).

Techniques: Fluorescence, Labeling

Journal: BMC Microbiology

Article Title: Localization of Staphylococcus aureus in tissue from the nasal vestibule in healthy carriers

doi: 10.1186/s12866-017-0997-3

Figure Lengend Snippet: Confocal laser scanning microscopy of frozen section of nasal tissue from an S. aureus carrier. Overview ( a-b) and detailed/zoomed portion of image ( c-d) showing S. aureus in stratum spinosum. Scale bar is in micrometers. Green fluorescence labeling of S. aureus , blue fluorescence labeling of keratinocyte cell nuclei and S. aureus DNA, red fluorescence labeling of actin. Primary antibody rabbit polyclonal antibody to S. aureus (ab20920, Abcam), secondary antibody Alexa Fluor 488® goat anti-rabbit IgG (Molecular Probe™, Thermo Fisher Scientific), Alexa Fluor 594 Phalloidin (Molecular Probes™, Thermo Fisher Scientific) and DRAQ5 (BioStatus). We used a CLSM ×63 objective

Article Snippet: S. aureus is labeled with primary rabbit polyclonal antibody to S. aureus (Abcam), secondary antibody Alexa Fluor 488 goat anti-rabbit IgG (green) (Molecular ProbesTM, Thermo Fisher Scientific), DRAQ5 (BioStatus) for keratinocyte nuclei (blue) and Alexa Fluor 594 Phalloidin (A12381, Molecular ProbesTM; Thermo Fisher Scientific) for actin (red).

Techniques: Confocal Laser Scanning Microscopy, Fluorescence, Labeling

Journal: BMC Microbiology

Article Title: Localization of Staphylococcus aureus in tissue from the nasal vestibule in healthy carriers

doi: 10.1186/s12866-017-0997-3

Figure Lengend Snippet: Localization of S. aureus on the border between stratum spinosum and stratum basale. Overview ( a) and detailed image ( b) of nasal tissue from an S. aureus carrier by confocal laser scanning microscopy. Scale bar is in micrometers. White brace indicates the region which is magnified in (b). Green fluorescence labeling of S. aureus , blue fluorescence labeling of keratinocyte cell nuclei and S. aureus DNA. Primary antibody rabbit polyclonal antibody to S. aureus (ab20920, Abcam), secondary antibody Alexa Fluor 488® goat anti-rabbit IgG (Molecular Probe™, Thermo Fisher Scientific), and DRAQ5 (BioStatus). We used a CLSM ×63 objective

Article Snippet: S. aureus is labeled with primary rabbit polyclonal antibody to S. aureus (Abcam), secondary antibody Alexa Fluor 488 goat anti-rabbit IgG (green) (Molecular ProbesTM, Thermo Fisher Scientific), DRAQ5 (BioStatus) for keratinocyte nuclei (blue) and Alexa Fluor 594 Phalloidin (A12381, Molecular ProbesTM; Thermo Fisher Scientific) for actin (red).

Techniques: Confocal Laser Scanning Microscopy, Fluorescence, Labeling

Journal: BMC Microbiology

Article Title: Localization of Staphylococcus aureus in tissue from the nasal vestibule in healthy carriers

doi: 10.1186/s12866-017-0997-3

Figure Lengend Snippet: Localization of S. aureus in the lower part of epidermis by CLSM. Overview and detailed image/zoomed portion of the image showing ( a ) S. aureus on the border between stratum spinosum and stratum basale, and ( b ) S. aureus on the border between the stratum basale and dermis. Inset represents a zoomed portion of the image. Scale bar is in micrometers. Green fluorescence labeling of S. aureus , blue fluorescence labeling of keratinocyte cell nuclei and S. aureus DNA. Primary antibody rabbit polyclonal antibody to S. aureus (ab20920, Abcam), secondary antibody Alexa Fluor 488® goat anti-rabbit IgG (Molecular Probe™, Thermo Fisher Scientific), and DRAQ5 (BioStatus). We used a CLSM ×63 objective

Article Snippet: S. aureus is labeled with primary rabbit polyclonal antibody to S. aureus (Abcam), secondary antibody Alexa Fluor 488 goat anti-rabbit IgG (green) (Molecular ProbesTM, Thermo Fisher Scientific), DRAQ5 (BioStatus) for keratinocyte nuclei (blue) and Alexa Fluor 594 Phalloidin (A12381, Molecular ProbesTM; Thermo Fisher Scientific) for actin (red).

Techniques: Fluorescence, Labeling

Journal: BMC Microbiology

Article Title: Localization of Staphylococcus aureus in tissue from the nasal vestibule in healthy carriers

doi: 10.1186/s12866-017-0997-3

Figure Lengend Snippet: Intracellular localization of S. aureus in nasal epithelial cells. S. aureus is labeled with primary rabbit polyclonal antibody to S. aureus (Abcam), secondary antibody Alexa Fluor 488 goat anti-rabbit IgG (green) (Molecular Probes™, Thermo Fisher Scientific), DRAQ5 (BioStatus) for keratinocyte nuclei (blue) and Alexa Fluor 594 Phalloidin (A12381, Molecular Probes™; Thermo Fisher Scientific) for actin (red). Confocal laser scanning microscopy of frozen sections. Projection is constructed from confocal Z-stacks (0,2 um thick), 63× objective. Image to the left and on top corresponds to a vertical view in the z-plane. Z-plane images reveal a single cellular nucleus (blue) closely related to fluorescing S. aureus (green)

Article Snippet: S. aureus is labeled with primary rabbit polyclonal antibody to S. aureus (Abcam), secondary antibody Alexa Fluor 488 goat anti-rabbit IgG (green) (Molecular ProbesTM, Thermo Fisher Scientific), DRAQ5 (BioStatus) for keratinocyte nuclei (blue) and Alexa Fluor 594 Phalloidin (A12381, Molecular ProbesTM; Thermo Fisher Scientific) for actin (red).

Techniques: Labeling, Confocal Laser Scanning Microscopy, Construct

Journal: Scientific Reports

Article Title: Development of the membrane ceiling method for in vitro spermatogenesis

doi: 10.1038/s41598-024-84965-1

Figure Lengend Snippet: Optimization of porous membranes used for the MC chip. ( A ) Representative images using an inverted microscope (BZ-X700) of the MC chip made of various porous membranes as indicated. Scale bar = 0.5 mm. ( B ) The ratio of tissue volume expansion and ( C ) the GFP-positive rate over 4 weeks with MC chips consisting of indicated membranes. ** P < 0.01 and P > 0.05 if no indication. Twelve (Cont), 8 (PC0.4), 8 (PC10), 8 (PET0.45), and 8 (PET3) tissues were used for the experiments. Images were taken by IX73 for quantifications.

Article Snippet: In brief, bright field and GFP images were taken by inverted microscopes with a tiling function (BioStation CT; Nikon, Tokyo, Japan, BZ-X700; Keyence Corporation, Osaka, Japan).

Techniques: Inverted Microscopy

Journal: Scientific Reports

Article Title: Development of the membrane ceiling method for in vitro spermatogenesis

doi: 10.1038/s41598-024-84965-1

Figure Lengend Snippet: Optimization of oxygen-permeable materials for device base. ( A ) Representative images showing the visibility of seminiferous tubules collected from GARN adult mice in the device using PMP or FEP. Images were taken by an inverted microscope (BZ-X700). Scale bar = 100 μm. ( B ) The ratio of tissue area expansion and ( C ) the GFP-positive rate for 5 weeks in the device with PDMS, PMP, or FEP base plate or dish. * P < 0.05, **** P < 0.0001, and ns; no significant. ( D ) Representative images of He-PAS staining of testis sections after culturing in the device with PDMS, PMP, or FEP base plate or dish. rST round spermatids, eST elongating spermatids. Scale bar = 200 μm (upper panel) and 50 μm (lower panel). ( E ) Live imaging of in vitro spermatogenesis of the same tubule during cultivation on the PMP bottom plate. The images were taken by BioStation CT. St step of spermatids. d culture days. Scale bar = 50 μm and 10 μm (zoomed in). ( F ) The number of mCherry-positive cells divided by tissue area. Each dot shows the average cell number per tissue area of frames. Images were taken at 5 weeks in the device with PDMS, PMP, and FEP using the tiling function of a microscope (BioStation CT). ** P < 0.01, **** P < 0.0001, and ns; no significant. Seventeen (PDMS), 17 (PMP), 8 (FEP) tissues were used for quantification.

Article Snippet: In brief, bright field and GFP images were taken by inverted microscopes with a tiling function (BioStation CT; Nikon, Tokyo, Japan, BZ-X700; Keyence Corporation, Osaka, Japan).

Techniques: Inverted Microscopy, Staining, Imaging, In Vitro, Microscopy

Journal: Scientific Reports

Article Title: Development of the membrane ceiling method for in vitro spermatogenesis

doi: 10.1038/s41598-024-84965-1

Figure Lengend Snippet: The effect of advanced DMEM/F12 (AD) as the in vitro basal medium in spermatogenesis. ( A ) The ratio of tissue area expansion and ( B ) the GFP-positive rate for 5 weeks in MEMα or AD-based culture medium. There were no significant differences ( P > 0.05). ( C ) Representative images of mCherry-positive cells observed using an inverted microscope (BZ-X700) after 5 weeks cultivated in MEMα or AD-based culture medium. Scale bar = 50 μm. ( D ) The number of mCherry-positive cells divided by tissue area (mm 2 ). Each dot shows the average cell number per tissue area of frames. Images were taken at 5 weeks using the tiling function of a microscope (BioStation CT and BZ-X700). * P < 0.05. Fifteen (MEMα) and 18 (AD) tissues were used for quantification. ( E ) In vitro generated sperm after 5 weeks in AD-based culture medium. The images were taken using an upright microscope (BX53). Flagellated cells with normal (upper panel) or abnormal (lower panel) heads were used for ICSI. Scale bar = 20 μm. ( F ) A picture of obtained offspring by ICSI using in vitro generated sperm.

Article Snippet: In brief, bright field and GFP images were taken by inverted microscopes with a tiling function (BioStation CT; Nikon, Tokyo, Japan, BZ-X700; Keyence Corporation, Osaka, Japan).

Techniques: In Vitro, Inverted Microscopy, Microscopy, Generated

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Temporal Heterogeneity Metrics in Apoptosis Induced by Anticancer Drugs

doi: 10.1369/0022155415583534

Figure Lengend Snippet: Early (A) and late (B) apoptosis in HeLa cells. Left column: DIC image. Right column: Overlay of four fluorescence channels in the same field of view. Hoechst 33342, blue; caspase 3/7 substrate, green, TMRE, red and DRAQ7, magenta. Early in apoptosis, caspase activity could be determined only in rounded cells. DRAQ7-positive (i.e., dead) cells could be seen only in late apoptosis. Notice the large number of inflated cells (arrowheads) with a high level of caspase 3/7 expression. Time (in hr) and treatment is given in the upper left corner. Scale, 20 μm. Images were taken with Planapo ×40/1.3 objective.

Article Snippet: HeLa cells stained with different fluorescent probes (TMRE, caspase 3/7 substrate, Hoechst 33342, Annexin V-Alexa Fluor 647, Cell ROX Deep Red and Sytox Blue (SB) from Life Technologies; DRAQ5 and DRAQ7 from Biostatus; Shepshed, UK; Asante Green AM (Asante NaTRIUM Green-2 AM) from Abcam, Cambridge, MA) or expressing Smac-DIABLO-GFP were imaged in a 37°C chamber on a Zeiss Axiovert 200 fluorescence microscope (Carl Zeiss Microscopy; Thornwood, NY) equipped with a CoolSNAP HQ2 camera (Photometrics; Tucson, AZ) and using SlideBook 5.0 software (Intelligent Imaging Innovations Inc., Denver, CO).

Techniques: Fluorescence, Activity Assay, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: LUZP1 Controls Cell Division, Migration and Invasion Through Regulation of the Actin Cytoskeleton

doi: 10.3389/fcell.2021.624089

Figure Lengend Snippet: Luzp1 –/– cells exhibit lower viability and higher apoptosis than WT. (A) Graphical representation of fold change in cell numbers of WT ( n = 5; blue line), Luzp1 –/– ( n = 5; orange line) and + LUZP1 cells ( n = 5; green line). P -values were calculated using Two-way ANOVA and Sidak’s multiple comparisons test. (B) Graphical representation of the percentage of WT (blue dots) and Luzp1 –/– cells (orange dots) in G0/G1, S or G2/M phases analyzed by EdU labeling and QIBC. P -values were calculated using Two-way ANOVA. (C) Graphical representation of the results of FACS analysis in using Annexin V and DRAQ7 staining to determine the percentage of apoptotic cells in WT and Luzp1 –/– cells in ( n = 4). P -value was calculated using Mann Whitney- U test. (D) Representative micrographs showing WT and Luzp1 –/– cells stained with the marker of mid-stage apoptosis cleaved caspase-3 (CC3) (green), phalloidin (magenta), and DAPI (blue). Black and white images show the single green and blue channels. Images were detected using Zeiss fluorescence microscope (Axio Imager D1), × 40 objective. Scale bar: 25 μm. (E) Graphical representation of quantification of CC3-positive cells in (D) showing the percentage of apoptotic cells in WT and Luzp1 –/– cells ( n = 20 micrographs, objective 20x). P -value was calculated using Mann Whitney- U test. The graphs in (A–C,E) represent the Mean and SEM. ∗ P < 0.05; ∗∗∗ P < 0.001; **** P < 0.0001.

Article Snippet: To evaluate apoptosis, Shh-LIGHT2 cells were washed with 1x PBS and then stained with Annexin V (BD Biosciences) and DRAQ7 (Biostatus Ltd.).

Techniques: Labeling, Staining, MANN-WHITNEY, Marker, Fluorescence, Microscopy