calcein Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore fluorexon calcein
    Fluorescence imaging of intact large PEGylated liposomes in the peritoneal cavity. Detection of intact liposomes in the peritoneum came from using fluorescence imaging of i.p.-administered <t>calcein-containing</t> zwitterionic liposomes. Mice were sacrificed
    Fluorexon Calcein, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 385 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fluorexon calcein/product/Millipore
    Average 99 stars, based on 385 article reviews
    Price from $9.99 to $1999.99
    fluorexon calcein - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    99
    Millipore calcein acetoxymethylester
    Fluorescence imaging of intact large PEGylated liposomes in the peritoneal cavity. Detection of intact liposomes in the peritoneum came from using fluorescence imaging of i.p.-administered <t>calcein-containing</t> zwitterionic liposomes. Mice were sacrificed
    Calcein Acetoxymethylester, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/calcein acetoxymethylester/product/Millipore
    Average 99 stars, based on 30 article reviews
    Price from $9.99 to $1999.99
    calcein acetoxymethylester - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    Image Search Results


    Fluorescence imaging of intact large PEGylated liposomes in the peritoneal cavity. Detection of intact liposomes in the peritoneum came from using fluorescence imaging of i.p.-administered calcein-containing zwitterionic liposomes. Mice were sacrificed

    Journal: Journal of liposome research

    Article Title: Large anti-HER2/neu liposomes for potential targeted intraperitoneal therapy of micrometastatic cancer

    doi: 10.3109/08982100903544185

    Figure Lengend Snippet: Fluorescence imaging of intact large PEGylated liposomes in the peritoneal cavity. Detection of intact liposomes in the peritoneum came from using fluorescence imaging of i.p.-administered calcein-containing zwitterionic liposomes. Mice were sacrificed

    Article Snippet: Cholesterol, phosphate-buffered saline (PBS), fluorexon (calcein), Sephadex G-50, diethylenetriaminepentaacetic acid (DTPA), 8-hydroxyquinoline (oxine), ascorbic acid, Triton X-100, and the control ascites fluid from murine myeloma were purchased from Sigma-Aldrich (St. Louis, Missouri, USA).

    Techniques: Fluorescence, Imaging, Mouse Assay

    IL-12p40 −/− mice increased bone formation and inhibited bone resorption. ( a ) Cytokine expression in different time periods after implantation in C57BL/6J mice. n =5 per group. ( b ) Increased cortical bone thickness and trabecular bone in micro-CT images of the distal femur of IL-12p40 −/− mice compared with WT mice. n =5 per group. ( c–f ) BMD ( c ), BV/TV ( d ), Tb.Th ( e ), and Tb.Sp ( f ) as measured by micro-CT. n =5 per group. ( g ) H E staining of femur sections from 8-week-old WT and IL-12p40 −/− mice. n =5 per group. Scale bars, 200 μ m. ( h ) ALP staining and quantitative analysis by ImageJ showed a more osteoblast surface in the IL-12p40 −/− mice compared with the WT mice. n =5 per group. Scale bars, 100 μ m. ( i ) TRAP staining and quantitative analysis by ImageJ showed that osteoclast surface was reduced in the IL-12p40 −/− mice compared with the WT mice. n =5 per group. Scale bars, 100 μ m. ( j and k ) ELISA of serum concentrations of OCN ( j ) and CTX-1 ( k ) in 8-week-old WT versus IL-12p40 −/− mice. n =5–8 per group. ( l ) Representative images (left) of new bone formation and quantification of the MAR (right) as assessed by double calcein labeling. n =5 per group. Scale bars, 25 μ m. All values are given as the mean±S.D. of three independent experiments. * P

    Journal: Cell Death and Differentiation

    Article Title: IL-12p40 impairs mesenchymal stem cell-mediated bone regeneration via CD4+ T cells

    doi: 10.1038/cdd.2016.72

    Figure Lengend Snippet: IL-12p40 −/− mice increased bone formation and inhibited bone resorption. ( a ) Cytokine expression in different time periods after implantation in C57BL/6J mice. n =5 per group. ( b ) Increased cortical bone thickness and trabecular bone in micro-CT images of the distal femur of IL-12p40 −/− mice compared with WT mice. n =5 per group. ( c–f ) BMD ( c ), BV/TV ( d ), Tb.Th ( e ), and Tb.Sp ( f ) as measured by micro-CT. n =5 per group. ( g ) H E staining of femur sections from 8-week-old WT and IL-12p40 −/− mice. n =5 per group. Scale bars, 200 μ m. ( h ) ALP staining and quantitative analysis by ImageJ showed a more osteoblast surface in the IL-12p40 −/− mice compared with the WT mice. n =5 per group. Scale bars, 100 μ m. ( i ) TRAP staining and quantitative analysis by ImageJ showed that osteoclast surface was reduced in the IL-12p40 −/− mice compared with the WT mice. n =5 per group. Scale bars, 100 μ m. ( j and k ) ELISA of serum concentrations of OCN ( j ) and CTX-1 ( k ) in 8-week-old WT versus IL-12p40 −/− mice. n =5–8 per group. ( l ) Representative images (left) of new bone formation and quantification of the MAR (right) as assessed by double calcein labeling. n =5 per group. Scale bars, 25 μ m. All values are given as the mean±S.D. of three independent experiments. * P

    Article Snippet: Double calcein labeling was performed by intraperitoneal injection of calcein (10 μ g/g body weight; C0875; Sigma-Aldrich) at 10 and 3 days before euthanasia.

    Techniques: Mouse Assay, Expressing, Micro-CT, Staining, ALP Assay, Enzyme-linked Immunosorbent Assay, Labeling

    An observation initiated 5 h after the addition of calcein (24 h after the addition of Hym-248). (A) Confirmation of crystal (white arrows in the left image, z=1 µm from the bottom) and SCM distribution (red arrows in the right image, z=6 µm from the bottom). Scale bar: 20 µm. (B) Time-lapse image showing a series of SCM pockets contractile movements in the area denoted by a white line in (A) . The numbers in the upper left of each panel indicate the recording times. Red and white arrows indicate contraction and expansion of SCM pockets, respectively. Scale bar: 10 µm.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Calcification process dynamics in coral primary polyps as observed using a calcein incubation method

    doi: 10.1016/j.bbrep.2017.01.006

    Figure Lengend Snippet: An observation initiated 5 h after the addition of calcein (24 h after the addition of Hym-248). (A) Confirmation of crystal (white arrows in the left image, z=1 µm from the bottom) and SCM distribution (red arrows in the right image, z=6 µm from the bottom). Scale bar: 20 µm. (B) Time-lapse image showing a series of SCM pockets contractile movements in the area denoted by a white line in (A) . The numbers in the upper left of each panel indicate the recording times. Red and white arrows indicate contraction and expansion of SCM pockets, respectively. Scale bar: 10 µm.

    Article Snippet: 2.2 Calcein Calcein was purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques:

    An observation in calcein-containing filtered seawater (FSW-calcein) at 6 h after the addition of Hym-248. The numbers in the upper and bottom parts of the panels indicate the recording times. Dotted lines indicate the periphery of the attached bottom part of the coral primary polyp. (A) Time series of images showing the developmental process of the primary polyp at the bottom (coral skeletal growth: bright green); distribution of calcein (green); and coral tissues (black). White arrows indicate the initial crystallization (12–15 h). SCMs are indicated by yellow arrows. Scale bar: 100 µm. (B) High-magnification images of the square area denoted by a white line in (A) . Red arrows indicate the direction of skeletal growth along the narrow SCM pocket. White arrows indicate crystals without growth. Scale bar: 50 µm.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Calcification process dynamics in coral primary polyps as observed using a calcein incubation method

    doi: 10.1016/j.bbrep.2017.01.006

    Figure Lengend Snippet: An observation in calcein-containing filtered seawater (FSW-calcein) at 6 h after the addition of Hym-248. The numbers in the upper and bottom parts of the panels indicate the recording times. Dotted lines indicate the periphery of the attached bottom part of the coral primary polyp. (A) Time series of images showing the developmental process of the primary polyp at the bottom (coral skeletal growth: bright green); distribution of calcein (green); and coral tissues (black). White arrows indicate the initial crystallization (12–15 h). SCMs are indicated by yellow arrows. Scale bar: 100 µm. (B) High-magnification images of the square area denoted by a white line in (A) . Red arrows indicate the direction of skeletal growth along the narrow SCM pocket. White arrows indicate crystals without growth. Scale bar: 50 µm.

    Article Snippet: 2.2 Calcein Calcein was purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: Crystallization Assay

    Vertical distribution of SCM, floating crystals, and calicoblastic cells. (A) High-magnification 3-dimensional image of the bottom of the coral primary polyp 24 h after the addition of Hym-248. The vertical height of this image is 5 µm. This image highlights the bottom of the tissue. Yellow arrows indicate relatively larger SCM pockets. The black area represents calicoblastic cells. Scale bar: 10 µm. (B) High-magnification image of the area enclosed by the white dotted line in (A) . Red arrows indicate the FSW-calcein distribution inside the cell (black). The area encircled by the white dotted line indicates dark thread-like structures among the cells. Scale bar: 5 µm. (C) Three-dimensional image at 0–3 µm from the bottom of the polyp, at the same position as in (A) . Blue arrow indicates the directions of observation of the cross-section image in (E) . Scale bar: 10 µm. (D) High-magnification image of the area enclosed by the white dotted line in (C) . White arrows indicate smaller SCM pockets. The yellow arrow indicates a putative nascent crystal. The red arrow indicates SCM pockets that appears to be surrounded by a calicoblastic cell (enclosed by dotted white line). Scale bar: 5 µm. (E) Image cross-section of (C) . White arrows indicate putative nascent crystals. SCM distribution is visible as darker green areas. (F) Time series of high magnification images reveal the initial skeletal growth, with a focus on the outer to intermediate areas in a primary polyp. Observation began 18 h after Hym-248 addition. The white arrow indicates an emerging crystal. Scale bar: 5 µm. (G) White and red arrows indicate floating putative nascent crystals at the bottom of the coral tissue in the same position as in (F) . Scale bar: 5 µm.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Calcification process dynamics in coral primary polyps as observed using a calcein incubation method

    doi: 10.1016/j.bbrep.2017.01.006

    Figure Lengend Snippet: Vertical distribution of SCM, floating crystals, and calicoblastic cells. (A) High-magnification 3-dimensional image of the bottom of the coral primary polyp 24 h after the addition of Hym-248. The vertical height of this image is 5 µm. This image highlights the bottom of the tissue. Yellow arrows indicate relatively larger SCM pockets. The black area represents calicoblastic cells. Scale bar: 10 µm. (B) High-magnification image of the area enclosed by the white dotted line in (A) . Red arrows indicate the FSW-calcein distribution inside the cell (black). The area encircled by the white dotted line indicates dark thread-like structures among the cells. Scale bar: 5 µm. (C) Three-dimensional image at 0–3 µm from the bottom of the polyp, at the same position as in (A) . Blue arrow indicates the directions of observation of the cross-section image in (E) . Scale bar: 10 µm. (D) High-magnification image of the area enclosed by the white dotted line in (C) . White arrows indicate smaller SCM pockets. The yellow arrow indicates a putative nascent crystal. The red arrow indicates SCM pockets that appears to be surrounded by a calicoblastic cell (enclosed by dotted white line). Scale bar: 5 µm. (E) Image cross-section of (C) . White arrows indicate putative nascent crystals. SCM distribution is visible as darker green areas. (F) Time series of high magnification images reveal the initial skeletal growth, with a focus on the outer to intermediate areas in a primary polyp. Observation began 18 h after Hym-248 addition. The white arrow indicates an emerging crystal. Scale bar: 5 µm. (G) White and red arrows indicate floating putative nascent crystals at the bottom of the coral tissue in the same position as in (F) . Scale bar: 5 µm.

    Article Snippet: 2.2 Calcein Calcein was purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques:

    Confirmation of coral skeletons and tissue staining patterns with or without calcein. Specimens were incubated in calcein-containing seawater during the experiment. (A) Bright-field image of a primary polyp 12 h after incubation. Scale bar: 200 µm. (B) Confocal image of the same position in (A). (C) Bright-field image of the primary polyp in (A) at 56 h after incubation. The black area indicates the coral skeleton. Scale bar: 200 µm. (D) Confocal image of the same position in (C) . Dotted lines indicate the area approximately quarter to half from the periphery of the primary polyp. The coral skeleton was stained using calcein (green). (E) High-magnification bright-field image of coral skeletons from the polyps in (A–D) at 24 h after incubation. The white arrow indicates a dumbbell-shaped crystal. Scale bar: 20 µm. (F) Confocal image of the same position in (E) . White arrow indicates a crystal on the surface of the glass-based dish. Dotted lines indicate the periphery of the subcalicoblastic medium (SCM). The black area indicates the bottom of the coral tissue.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Calcification process dynamics in coral primary polyps as observed using a calcein incubation method

    doi: 10.1016/j.bbrep.2017.01.006

    Figure Lengend Snippet: Confirmation of coral skeletons and tissue staining patterns with or without calcein. Specimens were incubated in calcein-containing seawater during the experiment. (A) Bright-field image of a primary polyp 12 h after incubation. Scale bar: 200 µm. (B) Confocal image of the same position in (A). (C) Bright-field image of the primary polyp in (A) at 56 h after incubation. The black area indicates the coral skeleton. Scale bar: 200 µm. (D) Confocal image of the same position in (C) . Dotted lines indicate the area approximately quarter to half from the periphery of the primary polyp. The coral skeleton was stained using calcein (green). (E) High-magnification bright-field image of coral skeletons from the polyps in (A–D) at 24 h after incubation. The white arrow indicates a dumbbell-shaped crystal. Scale bar: 20 µm. (F) Confocal image of the same position in (E) . White arrow indicates a crystal on the surface of the glass-based dish. Dotted lines indicate the periphery of the subcalicoblastic medium (SCM). The black area indicates the bottom of the coral tissue.

    Article Snippet: 2.2 Calcein Calcein was purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: Staining, Incubation

    Alkaline phosphatase (ALP) activity and bone double-labeling of MPS during expansion. (A) ALP staining was performed on MPS after expansion. The ALP-positive area was stained dark purple. Quantification of the ALP-positive area was evaluated with NIH image software. (B) Bone histomorphometric analysis after expansion by the bone double-labeling method with tetracycline (Tet) and calcein (Cal). Mineral apposition rate (MAR), bone formation rate (BFR), and mineralizing surface/bone surface (MS/BS) for each group was evaluated on the toluidine blue staining images. Scale bar, 50 μm. Individual data from the subjects are represented by open circles (○) and median values are indicated by closed rhombi (♦), and n = 4 for each group. * p

    Journal: Bone Reports

    Article Title: Relaxin 2 carried by magnetically directed liposomes accelerates rat midpalatal suture expansion and subsequent new bone formation

    doi: 10.1016/j.bonr.2019.100202

    Figure Lengend Snippet: Alkaline phosphatase (ALP) activity and bone double-labeling of MPS during expansion. (A) ALP staining was performed on MPS after expansion. The ALP-positive area was stained dark purple. Quantification of the ALP-positive area was evaluated with NIH image software. (B) Bone histomorphometric analysis after expansion by the bone double-labeling method with tetracycline (Tet) and calcein (Cal). Mineral apposition rate (MAR), bone formation rate (BFR), and mineralizing surface/bone surface (MS/BS) for each group was evaluated on the toluidine blue staining images. Scale bar, 50 μm. Individual data from the subjects are represented by open circles (○) and median values are indicated by closed rhombi (♦), and n = 4 for each group. * p

    Article Snippet: Four rats of each group were intraperitoneally injected with tetracycline hydrochloride (20 mg/kg; Sigma-Aldrich) on day 0 and calcein (20 mg/kg; Sigma-Aldrich, St. Louis, MO, USA) on day 5 for bone double labeling ( A).

    Techniques: ALP Assay, Activity Assay, Labeling, Staining, Software, Mass Spectrometry

    Role of VCAM-1/α4β1 binding in monocyte adhesion to ccRCC. (A) Analysis of VCAM-1 protein levels in 786-O-pRv or 786-O-VHL cells untreated or transfected with a scrambled siRNA (Scr) or siRNAs specific for VCAM-1. α-Tubulin was used as a loading control. A Western blot representative of three experiments is shown. (B) U937–calcein-AM–labeled cell adhesion on 786-O-pRv or 786-O-VHL cells untreated or transfected with a scrambled siRNA or siRNAs specific for VCAM-1. Cell adhesion was performed for 20 min at 37°C, and afterward, fluorescent cells were counted under the microscope. n = 5. (C) U937–calcein-AM–labeled adhesion experiment on 786-O-pRv or 786-O-VHL untreated (control [Ctr]) or treated with 10 µg/ml of blocking antibodies against β1, α4, or αL integrin subunits. Cell adhesion was performed for 20 min at 37°C, and afterward, fluorescent cells were counted under the microscope. n = 4. (B and C) Data are represented as number of cells ± SEM. 10 random fields were analyzed per condition. Statistical analysis was done using two-way ANOVA followed by Bonferroni’s posthoc test. (D) Effects of VHL loss in monocytic cell-mediated cytotoxicity against ccRCC cells. 786-O-pRv or 786-O-VHL target cells were seeded into 16-well sensor plates, and activated human monocytes treated with IFN-γ and LPS were directly added into wells at a 60:1 ratio, monocytes (Mono)/ccRCCs. Cell survival measurements were automatically collected every 5 min by the real-time cell electronic-sensing analyzer system (xCELLigence System) for up to 96 h. Cellular index results were expressed as cell survival percentage mean ± SEM at 96 h. Data are representative of three different experiments. Statistical analysis between different conditions was done using two-way ANOVA followed by Bonferroni’s posthoc test. **, P

    Journal: The Journal of Cell Biology

    Article Title: VHL promotes immune response against renal cell carcinoma via NF-κB–dependent regulation of VCAM-1

    doi: 10.1083/jcb.201608024

    Figure Lengend Snippet: Role of VCAM-1/α4β1 binding in monocyte adhesion to ccRCC. (A) Analysis of VCAM-1 protein levels in 786-O-pRv or 786-O-VHL cells untreated or transfected with a scrambled siRNA (Scr) or siRNAs specific for VCAM-1. α-Tubulin was used as a loading control. A Western blot representative of three experiments is shown. (B) U937–calcein-AM–labeled cell adhesion on 786-O-pRv or 786-O-VHL cells untreated or transfected with a scrambled siRNA or siRNAs specific for VCAM-1. Cell adhesion was performed for 20 min at 37°C, and afterward, fluorescent cells were counted under the microscope. n = 5. (C) U937–calcein-AM–labeled adhesion experiment on 786-O-pRv or 786-O-VHL untreated (control [Ctr]) or treated with 10 µg/ml of blocking antibodies against β1, α4, or αL integrin subunits. Cell adhesion was performed for 20 min at 37°C, and afterward, fluorescent cells were counted under the microscope. n = 4. (B and C) Data are represented as number of cells ± SEM. 10 random fields were analyzed per condition. Statistical analysis was done using two-way ANOVA followed by Bonferroni’s posthoc test. (D) Effects of VHL loss in monocytic cell-mediated cytotoxicity against ccRCC cells. 786-O-pRv or 786-O-VHL target cells were seeded into 16-well sensor plates, and activated human monocytes treated with IFN-γ and LPS were directly added into wells at a 60:1 ratio, monocytes (Mono)/ccRCCs. Cell survival measurements were automatically collected every 5 min by the real-time cell electronic-sensing analyzer system (xCELLigence System) for up to 96 h. Cellular index results were expressed as cell survival percentage mean ± SEM at 96 h. Data are representative of three different experiments. Statistical analysis between different conditions was done using two-way ANOVA followed by Bonferroni’s posthoc test. **, P

    Article Snippet: THP-1 or U937 monocytic cell lines from ATCC were labeled with the fluorescent dye calcein AM (2.5 mM; 148504-34-1; Sigma-Aldrich) in serum-free medium for 20 min at 37°C.

    Techniques: Binding Assay, Transfection, Western Blot, Labeling, Microscopy, Blocking Assay

    Effects of VHL loss and hypoxia on monocytic cell adhesion to ccRCC cells. (A and B) 786-O-VHL or 786-pRv cells were grown at confluence in a 24-multiwell plate and cultured under normoxic conditions. Then, THP1 (A) or U937 (B) monocytic cell lines (60 × 10 3 cells/well) previously labeled with 10 mM calcein-AM (green) were added. Cell adhesion was performed for 20 min at 37°C, and afterward, attached fluorescent cells were counted under the microscope. Two-tailed Student’s t test was performed. Representative images of attached THP1 or U937 monocytic cell (green) are shown. Bars, 50 μm. (C) U937–calcein-AM–labeled cell adhesion on 786-O-VHL or 786-O-pRv cells previously cultured under normoxia (Nx) or hypoxia 0.5% O 2 (Hp) for 24 h. Cell adhesion was performed for 20 min at 37°C, and afterward, fluorescent cells were counted under the microscope. Statistical analysis was done using two-way ANOVA followed by Bonferroni’s posthoc test. Data are represented as number of cells ± SEM. n = 5. 10 random fields were analyzed per condition. *, P

    Journal: The Journal of Cell Biology

    Article Title: VHL promotes immune response against renal cell carcinoma via NF-κB–dependent regulation of VCAM-1

    doi: 10.1083/jcb.201608024

    Figure Lengend Snippet: Effects of VHL loss and hypoxia on monocytic cell adhesion to ccRCC cells. (A and B) 786-O-VHL or 786-pRv cells were grown at confluence in a 24-multiwell plate and cultured under normoxic conditions. Then, THP1 (A) or U937 (B) monocytic cell lines (60 × 10 3 cells/well) previously labeled with 10 mM calcein-AM (green) were added. Cell adhesion was performed for 20 min at 37°C, and afterward, attached fluorescent cells were counted under the microscope. Two-tailed Student’s t test was performed. Representative images of attached THP1 or U937 monocytic cell (green) are shown. Bars, 50 μm. (C) U937–calcein-AM–labeled cell adhesion on 786-O-VHL or 786-O-pRv cells previously cultured under normoxia (Nx) or hypoxia 0.5% O 2 (Hp) for 24 h. Cell adhesion was performed for 20 min at 37°C, and afterward, fluorescent cells were counted under the microscope. Statistical analysis was done using two-way ANOVA followed by Bonferroni’s posthoc test. Data are represented as number of cells ± SEM. n = 5. 10 random fields were analyzed per condition. *, P

    Article Snippet: THP-1 or U937 monocytic cell lines from ATCC were labeled with the fluorescent dye calcein AM (2.5 mM; 148504-34-1; Sigma-Aldrich) in serum-free medium for 20 min at 37°C.

    Techniques: Cell Culture, Labeling, Microscopy, Two Tailed Test

    Illustrations of small micromere development from birth during cleavage stages through coelomic pouch residence in pluteus larva. Each stage of small micromere development can be labeled with small molecule dyes such as calcein-AM or nucleoside analogs like EdU. Later stages of small micromere development, including migration behaviors and left / right coelomic pouch distributions, can be observed after injecting RNAs encoding fluorescently tagged germline-specific genes or by engineering messages to contain the Nanos2 UTR retention sequences.

    Journal: Methods in cell biology

    Article Title: Methods to label, isolate, and image sea urchin small micromeres, the primordial germ cells (PGCs)

    doi: 10.1016/bs.mcb.2018.11.007

    Figure Lengend Snippet: Illustrations of small micromere development from birth during cleavage stages through coelomic pouch residence in pluteus larva. Each stage of small micromere development can be labeled with small molecule dyes such as calcein-AM or nucleoside analogs like EdU. Later stages of small micromere development, including migration behaviors and left / right coelomic pouch distributions, can be observed after injecting RNAs encoding fluorescently tagged germline-specific genes or by engineering messages to contain the Nanos2 UTR retention sequences.

    Article Snippet: In this case, the free fluorescent calcein (Sigma, C-0875) is dissolved in DMSO at a concentration of 1 mM and diluted in FSW to solutions of 125 μM to 30 nM.

    Techniques: Labeling, Migration

    Small molecule dyes are retained in small micromeres of multiple euechinoid species. (A) Calcein-AM, Celltrace RedOrange (CTRO), and Bo-Dipy-FL-Verapamil (BFLVp) and Vinblastine (BFLVb) are retained in small micromeres of blastula stage embryos of (B) Lytechinus pictus and Dendraster excentricus .

    Journal: Methods in cell biology

    Article Title: Methods to label, isolate, and image sea urchin small micromeres, the primordial germ cells (PGCs)

    doi: 10.1016/bs.mcb.2018.11.007

    Figure Lengend Snippet: Small molecule dyes are retained in small micromeres of multiple euechinoid species. (A) Calcein-AM, Celltrace RedOrange (CTRO), and Bo-Dipy-FL-Verapamil (BFLVp) and Vinblastine (BFLVb) are retained in small micromeres of blastula stage embryos of (B) Lytechinus pictus and Dendraster excentricus .

    Article Snippet: In this case, the free fluorescent calcein (Sigma, C-0875) is dissolved in DMSO at a concentration of 1 mM and diluted in FSW to solutions of 125 μM to 30 nM.

    Techniques:

    Elevated osteoclast number in IL-1rKO mice. Shown is a histomorphometric analysis. ( A ) Osteoclast number, surrogate of bone resorption activity, calculated as the number of TRAP-positive osteoclasts per trabecular surface area. ( B - D ) Measurements based on fluorescent calcein labeling of newly formed bone. ( B ) BFR, surrogate of total bone formation activity, calculated as the arithmetic product of MAR and Min.Peri. ( C ) Min.Peri, surrogate of osteoblast number, calculated as the percentage of calcein-labeled trabecular surface. ( D ) MAR, surrogate of osteoblast activity, calculated as the average distance between two calcein-labeled lines divided by the time interval between the two calcein injections. Data are mean ± SE in nine animals per condition. * , t test; P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Central IL-1 receptor signaling regulates bone growth and mass

    doi: 10.1073/pnas.0502562102

    Figure Lengend Snippet: Elevated osteoclast number in IL-1rKO mice. Shown is a histomorphometric analysis. ( A ) Osteoclast number, surrogate of bone resorption activity, calculated as the number of TRAP-positive osteoclasts per trabecular surface area. ( B - D ) Measurements based on fluorescent calcein labeling of newly formed bone. ( B ) BFR, surrogate of total bone formation activity, calculated as the arithmetic product of MAR and Min.Peri. ( C ) Min.Peri, surrogate of osteoblast number, calculated as the percentage of calcein-labeled trabecular surface. ( D ) MAR, surrogate of osteoblast activity, calculated as the average distance between two calcein-labeled lines divided by the time interval between the two calcein injections. Data are mean ± SE in nine animals per condition. * , t test; P

    Article Snippet: Four days and 1 day before killing, the animals were given the fluorochrome calcein intraperitoneally (15 mg/kg) (Sigma).

    Techniques: Mouse Assay, Activity Assay, Labeling

    Synergistic inhibitory effect of gallic acid and calycosin on LTB4-induced chemotaxis of neutrophils. (a) Schematic representation of neutrophil chemotaxis assay. (b) Quantitation of neutrophil chemotaxis. Human neutrophils were Isolated and treated with gallic acid and calycosin, individually or in combination, at 37°C for 24 h. At the end of treatment, neutrophils were loaded with fluorescent probe Calcein AM. Chemotaxis of neutrophils from the upper chamber towards LTB4 in lower chamber was assayed as outlined in (a). Migrated neutrophils were quantitated by measuring the fluorescence intensity on a Zeiss fluorescence spectroscopy (Jena, Germany). Data were expressed as the mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey's test. ∗∗∗ p

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Plant Natural Products Calycosin and Gallic Acid Synergistically Attenuate Neutrophil Infiltration and Subsequent Injury in Isoproterenol-Induced Myocardial Infarction: A Possible Role for Leukotriene B4 12-Hydroxydehydrogenase?

    doi: 10.1155/2015/434052

    Figure Lengend Snippet: Synergistic inhibitory effect of gallic acid and calycosin on LTB4-induced chemotaxis of neutrophils. (a) Schematic representation of neutrophil chemotaxis assay. (b) Quantitation of neutrophil chemotaxis. Human neutrophils were Isolated and treated with gallic acid and calycosin, individually or in combination, at 37°C for 24 h. At the end of treatment, neutrophils were loaded with fluorescent probe Calcein AM. Chemotaxis of neutrophils from the upper chamber towards LTB4 in lower chamber was assayed as outlined in (a). Migrated neutrophils were quantitated by measuring the fluorescence intensity on a Zeiss fluorescence spectroscopy (Jena, Germany). Data were expressed as the mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey's test. ∗∗∗ p

    Article Snippet: For the assay of chemotaxis, neutrophils were labelled by 5 μ M Calcein AM (Sigma, St. Louis, MO, USA).

    Techniques: Chemotaxis Assay, Quantitation Assay, Isolation, Fluorescence, Spectroscopy

    Impairment of periosteal bone formation in truncated BMPR-IB transgenic mice. (A and B) The responsiveness of tg(Col-2.3) homozygous transgenic mice to BMP-2. BMP-2 (20 μg/kg/d, daily for 5 d) in 20 μl PBS containing 0.1% BSA were injected into mice subcutaneously, adjacent to the calvarial bones. Significant amounts of new woven bone, indicated by red arrows and a black arrow head, were formed on the periosteal surface of calvariae in wild-type mice receiving BMP-2, but not in homozygous transgenic mice receiving BMP-2. The existing bones were indicated by a black arrow (A). Similarly, BMP-2 (20 μg/kg/d, ×5d) was administered subcutaneously and Calcein and Alizarin red were injected 7 and 2 d before the animals were killed. The new bone formation during the two fluorochrome injection (5-d period) was observed in wild-type mice (indicated by white arrows and a black arrowhead) but not in tg(Col-2.3) mice (B). (C) Effects of TGFβ on periosteal bone formation. TGFβ (20 μg/kg/d, daily for 5 d) was injected subcutaneously over calvariae. Significant amounts of new woven bone, indicated by red arrows and a black arrowhead, were formed on the periosteal surface of calvariae in wild-type and homozygous transgenic mice. (D and E) In vitro bone formation assay. Calvariae were isolated from 4-d-old wild-type and tg(Col-2.3) mice and cultured for 7 d in BGJ medium. At the end of the incubation, calvariae were fixed and processed for histomorphometric analyses. Osteoblasts on the BS were indicated by red arrows and existing bones were indicated by yellow arrows (D). The thickness of calvariae were measured and presented in (E).

    Journal: The Journal of Cell Biology

    Article Title: Bone morphogenetic protein receptor signaling is necessary for normal murine postnatal bone formation

    doi: 10.1083/jcb.200109012

    Figure Lengend Snippet: Impairment of periosteal bone formation in truncated BMPR-IB transgenic mice. (A and B) The responsiveness of tg(Col-2.3) homozygous transgenic mice to BMP-2. BMP-2 (20 μg/kg/d, daily for 5 d) in 20 μl PBS containing 0.1% BSA were injected into mice subcutaneously, adjacent to the calvarial bones. Significant amounts of new woven bone, indicated by red arrows and a black arrow head, were formed on the periosteal surface of calvariae in wild-type mice receiving BMP-2, but not in homozygous transgenic mice receiving BMP-2. The existing bones were indicated by a black arrow (A). Similarly, BMP-2 (20 μg/kg/d, ×5d) was administered subcutaneously and Calcein and Alizarin red were injected 7 and 2 d before the animals were killed. The new bone formation during the two fluorochrome injection (5-d period) was observed in wild-type mice (indicated by white arrows and a black arrowhead) but not in tg(Col-2.3) mice (B). (C) Effects of TGFβ on periosteal bone formation. TGFβ (20 μg/kg/d, daily for 5 d) was injected subcutaneously over calvariae. Significant amounts of new woven bone, indicated by red arrows and a black arrowhead, were formed on the periosteal surface of calvariae in wild-type and homozygous transgenic mice. (D and E) In vitro bone formation assay. Calvariae were isolated from 4-d-old wild-type and tg(Col-2.3) mice and cultured for 7 d in BGJ medium. At the end of the incubation, calvariae were fixed and processed for histomorphometric analyses. Osteoblasts on the BS were indicated by red arrows and existing bones were indicated by yellow arrows (D). The thickness of calvariae were measured and presented in (E).

    Article Snippet: Tetracycline (15 mg/kg i.p.; Sigma-Aldrich) was administered to 1-mo-old wild-type and transgenic mice, and this was followed by injection of a calcein label (20 mg/kg i.p.; Sigma-Aldrich) 4 d later, mice were sacrificed 48 h after the second label was injected, and bone tissues were removed and fixed in 70% ethanol for 48 h. The specimens were dehydrated through a graded series of ethanol (70–100%) and embedded in methylmethacrylate without prior decalcification.

    Techniques: Transgenic Assay, Mouse Assay, Injection, In Vitro, Tube Formation Assay, Isolation, Cell Culture, Incubation

    Calcein release from liposome-microbubble complexes in response to ultrasound treatment. Calcein fluorescence (Mean±Standard Deviation) of untreated pendants (left bar), of ultrasound-treated pendants (center bar), and of completely ruptured liposomes following the addition of Triton X-100 (right bar).

    Journal: Journal of controlled release : official journal of the Controlled Release Society

    Article Title: Ultrasound-triggered release of materials entrapped in microbubble-liposome constructs: a tool for targeted drug delivery

    doi: 10.1016/j.jconrel.2010.07.115

    Figure Lengend Snippet: Calcein release from liposome-microbubble complexes in response to ultrasound treatment. Calcein fluorescence (Mean±Standard Deviation) of untreated pendants (left bar), of ultrasound-treated pendants (center bar), and of completely ruptured liposomes following the addition of Triton X-100 (right bar).

    Article Snippet: Briefly, lipid solution of phosphatidylcholine, cholesterol and biotin-amidocaproyl-phosphatidylethanolamine in chloroform and ethyl ether was mixed with the aqueous medium containing 0.2 M calcein dye (Sigma, St. Louis, MO) at the 3:1 volume ratio between organic and aqueous phase.

    Techniques: Fluorescence, Standard Deviation

    Inhibition of calcineurin (FK506-treatment) influences joint formation and regenerate length. (A, B) Calcein staining detects bone matrix and reveals joint failure and increased regenerate length in FK506-treated wild-type fish compared to control DMSO-treated wild-type fish. Panels on the left show fluorescence, while panels on the right show fluorescence plus bright field to better illustrate the end of the regenerating fin (the dotted line indicates the amputation plane). Arrowheads point to joints. Double-headed arrows identify regenerate length. (C) Regenerate length is significantly increased FK506-treated (n = 18) versus DMSO-treated (n = 18, negative control) fish. Statistically significant differences were determined by the student’s t-test where p

    Journal: Developmental dynamics : an official publication of the American Association of Anatomists

    Article Title: Cx43 suppresses evx1 expression to regulate joint initiation in the regenerating fin

    doi: 10.1002/dvdy.24531

    Figure Lengend Snippet: Inhibition of calcineurin (FK506-treatment) influences joint formation and regenerate length. (A, B) Calcein staining detects bone matrix and reveals joint failure and increased regenerate length in FK506-treated wild-type fish compared to control DMSO-treated wild-type fish. Panels on the left show fluorescence, while panels on the right show fluorescence plus bright field to better illustrate the end of the regenerating fin (the dotted line indicates the amputation plane). Arrowheads point to joints. Double-headed arrows identify regenerate length. (C) Regenerate length is significantly increased FK506-treated (n = 18) versus DMSO-treated (n = 18, negative control) fish. Statistically significant differences were determined by the student’s t-test where p

    Article Snippet: A 0.2% calcein solution was prepared by dissolving 2 g calcein powder (Sigma) in 1 L deionized water, and 0.5M NaOH was added to bring the pH to neutral.

    Techniques: Inhibition, Staining, Fluorescence In Situ Hybridization, Fluorescence, Negative Control

    In vitro hMSC viability and spreading within hydrogels. (A) Fluorescent image stacks of 200 μm of the outer and inner part of the hydrogels showing staining of the cytoplasm of live cells with calcein (green) and cell nuclei with Hoechst (blue) on day 1 and 3 after cell encapsulation. Scale bar: 100 μm. (B) Quantification of cell number within the hydrogels on day 1, 3 and 7 post-encapsulation. Data represent the mean of three experiments with the S.D. *p

    Journal: Acta biomaterialia

    Article Title: PEG Hydrogel Containing Calcium-Releasing Particles and Mesenchymal Stromal Cells Promote Vessel Maturation

    doi: 10.1016/j.actbio.2017.12.009

    Figure Lengend Snippet: In vitro hMSC viability and spreading within hydrogels. (A) Fluorescent image stacks of 200 μm of the outer and inner part of the hydrogels showing staining of the cytoplasm of live cells with calcein (green) and cell nuclei with Hoechst (blue) on day 1 and 3 after cell encapsulation. Scale bar: 100 μm. (B) Quantification of cell number within the hydrogels on day 1, 3 and 7 post-encapsulation. Data represent the mean of three experiments with the S.D. *p

    Article Snippet: Briefly, after being washed with Dulbecco’s phosphate-buffered saline (DPBS; Invitrogen), cell-containing hydrogels were incubated for 1 h at 37 °C in 5% (v/v) CO2 with 1 mL of non-supplemented αMEM containing 7.5 μM calcein reagent and 2.5 mM probenecid (Sigma-Aldrich).

    Techniques: In Vitro, Staining

    Postoperative fluorescence microphotograph (magnification ×100) at 7 weeks: Alizarin labels at 1 week (red, right-above), Calcein labels at 3 weeks (green, left-above), Oxytetracycline labels at 5 weeks (yellow, left-below) and merged images. The fluorescent images were taken around the center areas where new bone layer were found under Ti membranes.

    Journal: Journal of Tissue Engineering

    Article Title: In vivo bone regeneration by differently designed titanium membrane with or without surface treatment: a study in rat calvarial defects

    doi: 10.1177/2041731419831466

    Figure Lengend Snippet: Postoperative fluorescence microphotograph (magnification ×100) at 7 weeks: Alizarin labels at 1 week (red, right-above), Calcein labels at 3 weeks (green, left-above), Oxytetracycline labels at 5 weeks (yellow, left-below) and merged images. The fluorescent images were taken around the center areas where new bone layer were found under Ti membranes.

    Article Snippet: Red Alizarin complexone of 25 mg/kg (Sigma, St. Louis, MO, USA), green Calcein of 25 mg/kg (Sigma, St. Louis, MO, USA), and yellow Oxytetracycline of 20 mg/kg (Sigma, St. Louis, MO, USA) were administered at 1, 3, and 5 weeks after surgery by subcutaneous injection, respectively.

    Techniques: Fluorescence

    Mineralization dynamics of differentiating osteoblasts treated with osteogenic cocktail (OGC), detected by Calcein. a Fluorescent images of the cell culture obtained by IncuCyte ZOOM at day 7, day 10 and day 14 of differentiation. Last row: vehicle (ethanol) treated cells, showing no differentiation, representative pictures are shown, n = 3. b Mineralization curve generated automatically by the IncuCyte ZOOM based on the green object count per image. The cells were scanned automatically every 3 h for two weeks, representative experiment is shown, n = 3, SE

    Journal: Biological Procedures Online

    Article Title: Real-Time Vital Mineralization Detection and Quantification during In Vitro Osteoblast Differentiation

    doi: 10.1186/s12575-018-0079-4

    Figure Lengend Snippet: Mineralization dynamics of differentiating osteoblasts treated with osteogenic cocktail (OGC), detected by Calcein. a Fluorescent images of the cell culture obtained by IncuCyte ZOOM at day 7, day 10 and day 14 of differentiation. Last row: vehicle (ethanol) treated cells, showing no differentiation, representative pictures are shown, n = 3. b Mineralization curve generated automatically by the IncuCyte ZOOM based on the green object count per image. The cells were scanned automatically every 3 h for two weeks, representative experiment is shown, n = 3, SE

    Article Snippet: From powder calcein (catalogue N C0875 , Sigma-Aldrich) 10 mM calcein solution was prepared in 0.1 M of NaOH; it was further diluted 1:10 in ddH2 O in order to have a 1 mM working stock.

    Techniques: Cell Culture, Generated

    Comparison of quantification of mineralization obtained either by Alizarin Red ( a ) or calcein ( b ). MSCs were transfected by an siRNA against PPP2R2C and the mineralization was quantified on day 12 of osteoblast differentiation a Measurement of extracted Alizarin Red, b Measurement of calceinin cultures in the IncuCyte ZOOM. MSCs transfected with negative control siRNA were used as a control, n = 2, SE

    Journal: Biological Procedures Online

    Article Title: Real-Time Vital Mineralization Detection and Quantification during In Vitro Osteoblast Differentiation

    doi: 10.1186/s12575-018-0079-4

    Figure Lengend Snippet: Comparison of quantification of mineralization obtained either by Alizarin Red ( a ) or calcein ( b ). MSCs were transfected by an siRNA against PPP2R2C and the mineralization was quantified on day 12 of osteoblast differentiation a Measurement of extracted Alizarin Red, b Measurement of calceinin cultures in the IncuCyte ZOOM. MSCs transfected with negative control siRNA were used as a control, n = 2, SE

    Article Snippet: From powder calcein (catalogue N C0875 , Sigma-Aldrich) 10 mM calcein solution was prepared in 0.1 M of NaOH; it was further diluted 1:10 in ddH2 O in order to have a 1 mM working stock.

    Techniques: Transfection, Negative Control

    Comparison of the efficiency of calcein versus Alizarin Red to detect mineralized areas of osteoblast culture. a mineralization pattern detected by Olympus IX2-UCB Fluorescent microscope, phase-contrast (left panel, calcium hydroxyapatite deposits appear like yellowish structures at phase-contrast); the same section on the plate visualized by calcein by fluorescent setting (middle panel); overlap between the phase-contrast and fluorescent images in composite setting (right panel), scale bars 2000 μm. Representative pictures are shown, n = 3. b Calcein staining and Alizarin Red staining of the same mineralized areas detected by IncuCyte ZOOM: note that the phase-contrast objective of the IncuCyte ZOOM does not show the red color of Alizarin Red, however the pattern of the staining is clearly visible in gray scale and can be compared to the calceinpattern; representative pictures are shown, n = 3

    Journal: Biological Procedures Online

    Article Title: Real-Time Vital Mineralization Detection and Quantification during In Vitro Osteoblast Differentiation

    doi: 10.1186/s12575-018-0079-4

    Figure Lengend Snippet: Comparison of the efficiency of calcein versus Alizarin Red to detect mineralized areas of osteoblast culture. a mineralization pattern detected by Olympus IX2-UCB Fluorescent microscope, phase-contrast (left panel, calcium hydroxyapatite deposits appear like yellowish structures at phase-contrast); the same section on the plate visualized by calcein by fluorescent setting (middle panel); overlap between the phase-contrast and fluorescent images in composite setting (right panel), scale bars 2000 μm. Representative pictures are shown, n = 3. b Calcein staining and Alizarin Red staining of the same mineralized areas detected by IncuCyte ZOOM: note that the phase-contrast objective of the IncuCyte ZOOM does not show the red color of Alizarin Red, however the pattern of the staining is clearly visible in gray scale and can be compared to the calceinpattern; representative pictures are shown, n = 3

    Article Snippet: From powder calcein (catalogue N C0875 , Sigma-Aldrich) 10 mM calcein solution was prepared in 0.1 M of NaOH; it was further diluted 1:10 in ddH2 O in order to have a 1 mM working stock.

    Techniques: Microscopy, Staining

    Morphological changes and reduced bone content in the long bones of BgnFmod KO mice. ( a ) 3D µCT reconstruction of femora, showing the misalignment of the patellar groove with the long axis of the femoral shaft in the BgnFmod KO mice. Images are from median samples. WT femur from 11w old female. ( b ) Representative 2D axial projections of distal metaphysis of 32w old male WT and BgnFmod KO mice, showing the changes in morphology as well as the bony protuberance. Bar = 1 mm. ( c ) Representative histological section of calcein double labeling of distal femoral metaphysis of 11w old mice. Arrow pointing at bony lateral projection with dense trabecular architecture in the femur of BgnFmod KO mice. ( d ) Von-Kossa staining of 5w old femora presenting bony lateral projection and less mineralized bone in the BgnFmod KO mice compared with WT. Bar = 1 mm. ( e ) Bone mineral density (BMD) and (f) bone mineral content (BMC) of 11w old WT and BgnFmod KO mice. Obtained by DEXA analysis of whole body. Data are mean ± SE obtained from N = 5–7 mice per group, analyzed by unpaired, 2-tailed Student’s T test. *p

    Journal: Scientific Reports

    Article Title: Small leucine rich proteoglycans, a novel link to osteoclastogenesis

    doi: 10.1038/s41598-017-12651-6

    Figure Lengend Snippet: Morphological changes and reduced bone content in the long bones of BgnFmod KO mice. ( a ) 3D µCT reconstruction of femora, showing the misalignment of the patellar groove with the long axis of the femoral shaft in the BgnFmod KO mice. Images are from median samples. WT femur from 11w old female. ( b ) Representative 2D axial projections of distal metaphysis of 32w old male WT and BgnFmod KO mice, showing the changes in morphology as well as the bony protuberance. Bar = 1 mm. ( c ) Representative histological section of calcein double labeling of distal femoral metaphysis of 11w old mice. Arrow pointing at bony lateral projection with dense trabecular architecture in the femur of BgnFmod KO mice. ( d ) Von-Kossa staining of 5w old femora presenting bony lateral projection and less mineralized bone in the BgnFmod KO mice compared with WT. Bar = 1 mm. ( e ) Bone mineral density (BMD) and (f) bone mineral content (BMC) of 11w old WT and BgnFmod KO mice. Obtained by DEXA analysis of whole body. Data are mean ± SE obtained from N = 5–7 mice per group, analyzed by unpaired, 2-tailed Student’s T test. *p

    Article Snippet: Dynamic histomorphometry Prior to sacrifice WT and BgnFmod KO mice were injected with a calcein fluorochrome (Sigma-Aldrich, St. Louis, MO), 15 mg/Kg intraperitoneally.

    Techniques: Mouse Assay, Labeling, Staining

    Young BgnFmod KO mice have enhanced bone formation and overactive osteoblasts. ( a – c ) Dynamic histomorphometric parameters based on fluorescent visualization of calcein fluorochrome in trabecular compartment of distal metaphysis of femur. ( a ) Bone formation rate (BFR); ( b ) mineralizing perimeter (Min.Pm.); ( c ) Mineral appositional Rate (MAR); Data are mean ± SE obtained from N = 4–6 mice per group. ( d ) Osteocalcin serum levels. Serum was obtained from 5 w old mice before euthanization and measured using commercial ELISA kit. ( e – j ) In-vitro BMSC culture derived from 9 w old WT and BgnFmod KO mice. ( e ) Representative image of colony-forming efficiency showing the number of colonies initiated by a single colony-forming unit-fibroblast formed. ( f ) Enumeration of colonies shown in ( e ). Data are mean ± SE obtained from N = 3 cultures (3 different animals in 3 different cultures) per genotype. ( g ) Alizarin red S staining of BMSCs cultures grown in osteogenic medium for designated periods of time. ( h – j ) Quantification of the relative expression levels of osteogenic differentiation markers measured by real-time PCR in mRNA extracted from BgnFmod KO and WT BMSCs grown in osteogenic medium for up to 30d. ( h ) Osterix ( Ostx ); ( i ) alkaline phosphatase ( ALP ); ( j ) Osteocalcin ( Osc ). Data are mean ± SE obtained from N = 3 cultures obtained from 3 separate mice per genotype. *p

    Journal: Scientific Reports

    Article Title: Small leucine rich proteoglycans, a novel link to osteoclastogenesis

    doi: 10.1038/s41598-017-12651-6

    Figure Lengend Snippet: Young BgnFmod KO mice have enhanced bone formation and overactive osteoblasts. ( a – c ) Dynamic histomorphometric parameters based on fluorescent visualization of calcein fluorochrome in trabecular compartment of distal metaphysis of femur. ( a ) Bone formation rate (BFR); ( b ) mineralizing perimeter (Min.Pm.); ( c ) Mineral appositional Rate (MAR); Data are mean ± SE obtained from N = 4–6 mice per group. ( d ) Osteocalcin serum levels. Serum was obtained from 5 w old mice before euthanization and measured using commercial ELISA kit. ( e – j ) In-vitro BMSC culture derived from 9 w old WT and BgnFmod KO mice. ( e ) Representative image of colony-forming efficiency showing the number of colonies initiated by a single colony-forming unit-fibroblast formed. ( f ) Enumeration of colonies shown in ( e ). Data are mean ± SE obtained from N = 3 cultures (3 different animals in 3 different cultures) per genotype. ( g ) Alizarin red S staining of BMSCs cultures grown in osteogenic medium for designated periods of time. ( h – j ) Quantification of the relative expression levels of osteogenic differentiation markers measured by real-time PCR in mRNA extracted from BgnFmod KO and WT BMSCs grown in osteogenic medium for up to 30d. ( h ) Osterix ( Ostx ); ( i ) alkaline phosphatase ( ALP ); ( j ) Osteocalcin ( Osc ). Data are mean ± SE obtained from N = 3 cultures obtained from 3 separate mice per genotype. *p

    Article Snippet: Dynamic histomorphometry Prior to sacrifice WT and BgnFmod KO mice were injected with a calcein fluorochrome (Sigma-Aldrich, St. Louis, MO), 15 mg/Kg intraperitoneally.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay, In Vitro, Derivative Assay, Staining, Expressing, Real-time Polymerase Chain Reaction, ALP Assay