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  • 90
    Nikon nikon inverted fluorescent microscope
    Global nascent protein synthesis is repressed in Zscan4 + state cells and has elevated levels of Eif1a. (A) To determine global protein synthesis, mouse ES cells were cultured in methionine-free ES cell media for 30 min, labelled with AHA, a methionine analogue, for 1 h and visualized with Alexa Fluor 488 alkyne through Click-iT reaction and co-stained with Zscan4 antibody. Zscan + ES cells (i, arrowhead) showed dramatic repression in AHA staining (ii and iii). Eif1a levels were determined in Zscan4 + cells by co-staining with the respective antibodies. Zscan4 + cells (iv) showed enhanced Eif1a expression (v and vi). Pictures were taken at ×40 magnification using a Nikon Eclipse <t>TE300</t> microscope (scale bar indicates 50 mm). (B) pZscan4 -puro mouse ES cells were treated with (+) or without (−) 1.5 mg/ml puromycin for 1 or 2 days. Cells were labelled with AHA as described above and protein harvested using RIPA buffer containing protease inhibitors. AHA-labelled samples were then detected using TAMRA alkyne via Click-iT reaction. Samples were run on a 4–12% Bis-Tris SDS–PAGE acrylamide gel, and TAMRA was visualized under UV illuminescence. (C) Western blot analysis was performed on the same gel (C) to determine the levels of Zscan4 and Eif1a. Protein from the acrylamide gel was transferred onto the polyvinylidene difluoride (PVDF) membrane and blotted with antibodies against Zscan4, Eif1a, and beta-actin.
    Nikon Inverted Fluorescent Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 90/100, based on 363 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon te2000 nikon inverted fluorescence microscope
    Cd-induced extracellular Ca 2 + influx elevates [Ca 2 + ] i contributing to neuronal apoptosis via activation of MAPK and mTOR pathways. Indicated cells were pretreated with or without 100 µM EGTA for 30 min, and then exposed to Cd (10 and/or 20 µM) for 24 h (A–D) or 4 h (E, F). (A and B) [Ca 2+ ] i fluorescent intensities were evaluated as described in Materials and Methods . (C) Morphology of PC12 cells was assessed using a Nikon Eclipse <t>TE2000-U</t> inverted phase-contrast microscope (200×) equipped with digital camera. (D) Cell viability in SH-SY5Y cells and primary neurons was evaluated by one solution assay. (E and F) Indicated cell lysates were subjected to Western blotting using indicated antibodies. The blots were probed for β-tubulin as a loading control. Similar results were observed in at least three independent experiments. Results (A, D) are presented as mean ± SE; n = 6. b P
    Te2000 Nikon Inverted Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 77/100, based on 93 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    81
    Nikon inverted te300 nikon fluorescence microscope
    Cd-induced extracellular Ca 2 + influx elevates [Ca 2 + ] i contributing to neuronal apoptosis via activation of MAPK and mTOR pathways. Indicated cells were pretreated with or without 100 µM EGTA for 30 min, and then exposed to Cd (10 and/or 20 µM) for 24 h (A–D) or 4 h (E, F). (A and B) [Ca 2+ ] i fluorescent intensities were evaluated as described in Materials and Methods . (C) Morphology of PC12 cells was assessed using a Nikon Eclipse <t>TE2000-U</t> inverted phase-contrast microscope (200×) equipped with digital camera. (D) Cell viability in SH-SY5Y cells and primary neurons was evaluated by one solution assay. (E and F) Indicated cell lysates were subjected to Western blotting using indicated antibodies. The blots were probed for β-tubulin as a loading control. Similar results were observed in at least three independent experiments. Results (A, D) are presented as mean ± SE; n = 6. b P
    Inverted Te300 Nikon Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 81/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon te200 nikon inverted fluorescent microscope
    Establishment of an in vivo xenograft TAZ-overexpressing mouse model ( A ) Overexpression of TAZ-S89A in mouse immortalized lung epithelial cells (E10) caused highly malignant NSCLC tumor formation. Tumorigenesis assay was performed by subcutaneously injecting about 3 × 10 6 E10-TAZ-S89A cells into two-sides of nude mice. E10-TAZ-S89A cells caused large tumors (i) in two weeks. Two week later, the tumors were fixed, sectioned, and subjected to H E staining and IHC. H E staining with antibody incubation of E10-TAZ-S89A tumor section showed high-grade, poorly-differentiated carcinoma (ii). IHC staining for TAZ expression using TAZ antibody (1:300 dilution, BD Biosciences) showed that TAZ was overexpressed in the nuclei (iii). Pictures were taken using <t>TE200</t> Nikon Inverted Fluorescent Microscope (Nikon, Montreal, Canada) as 20× magnification. ( B ) Western blot analysis of TAZ-S89A expression. Ten μg of cell lysate extracted from E10-TAZS89A-P or E10-TAZ-S89A-T cells in the absence (−) or presence (+) of Dox were subjected to WB analysis using anti-TAZ (1:1000, BD Biosciences) and anti β-actin (1:10,000 Sigma, Oakville, Canada) antibodies. β-actin was used as an internal loading control. ( C ) Cell proliferation assay. Triplicates of 1.5 × 10 4 E10-TAZ-S89A-P or tumorigenic E10-TAZ-S89A-T cells were seeded into each well of 12-well plates, then untreated (−) or treated (+) with Dox. Cell numbers were counted on days 1, 2, 3, 4, 5, and 6 after plating. The experiments were repeated at least three times. Data are shown as means ± S.D. “*” represent significant difference. ( D – E ) Soft agar assay. Triplicates of 2×10 3 TAZ-S89A-P or E10-TAZ-S89A-T cells were mixed with 0.4% agarose in growth media and overlaid on 0.8% agarose in 6-well plates, followed by incubation in the absence or presence of Dox (2 μg/ml). Colony formation was examined after culturing for 18 days (D). Colony numbers were counted by Bio-Rad Gel Doc System (Bio-Rad, Mississauga, Canada). Data are shown as means ± S.D. “*” represent significant difference ( P
    Te200 Nikon Inverted Fluorescent Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 83/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon fluorescence microscopy a nikon ti e inverted fluorescence microscope
    Establishment of an in vivo xenograft TAZ-overexpressing mouse model ( A ) Overexpression of TAZ-S89A in mouse immortalized lung epithelial cells (E10) caused highly malignant NSCLC tumor formation. Tumorigenesis assay was performed by subcutaneously injecting about 3 × 10 6 E10-TAZ-S89A cells into two-sides of nude mice. E10-TAZ-S89A cells caused large tumors (i) in two weeks. Two week later, the tumors were fixed, sectioned, and subjected to H E staining and IHC. H E staining with antibody incubation of E10-TAZ-S89A tumor section showed high-grade, poorly-differentiated carcinoma (ii). IHC staining for TAZ expression using TAZ antibody (1:300 dilution, BD Biosciences) showed that TAZ was overexpressed in the nuclei (iii). Pictures were taken using <t>TE200</t> Nikon Inverted Fluorescent Microscope (Nikon, Montreal, Canada) as 20× magnification. ( B ) Western blot analysis of TAZ-S89A expression. Ten μg of cell lysate extracted from E10-TAZS89A-P or E10-TAZ-S89A-T cells in the absence (−) or presence (+) of Dox were subjected to WB analysis using anti-TAZ (1:1000, BD Biosciences) and anti β-actin (1:10,000 Sigma, Oakville, Canada) antibodies. β-actin was used as an internal loading control. ( C ) Cell proliferation assay. Triplicates of 1.5 × 10 4 E10-TAZ-S89A-P or tumorigenic E10-TAZ-S89A-T cells were seeded into each well of 12-well plates, then untreated (−) or treated (+) with Dox. Cell numbers were counted on days 1, 2, 3, 4, 5, and 6 after plating. The experiments were repeated at least three times. Data are shown as means ± S.D. “*” represent significant difference. ( D – E ) Soft agar assay. Triplicates of 2×10 3 TAZ-S89A-P or E10-TAZ-S89A-T cells were mixed with 0.4% agarose in growth media and overlaid on 0.8% agarose in 6-well plates, followed by incubation in the absence or presence of Dox (2 μg/ml). Colony formation was examined after culturing for 18 days (D). Colony numbers were counted by Bio-Rad Gel Doc System (Bio-Rad, Mississauga, Canada). Data are shown as means ± S.D. “*” represent significant difference ( P
    Fluorescence Microscopy A Nikon Ti E Inverted Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 78/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon fluorescence inverse microscope nikon eclipse80i
    Establishment of an in vivo xenograft TAZ-overexpressing mouse model ( A ) Overexpression of TAZ-S89A in mouse immortalized lung epithelial cells (E10) caused highly malignant NSCLC tumor formation. Tumorigenesis assay was performed by subcutaneously injecting about 3 × 10 6 E10-TAZ-S89A cells into two-sides of nude mice. E10-TAZ-S89A cells caused large tumors (i) in two weeks. Two week later, the tumors were fixed, sectioned, and subjected to H E staining and IHC. H E staining with antibody incubation of E10-TAZ-S89A tumor section showed high-grade, poorly-differentiated carcinoma (ii). IHC staining for TAZ expression using TAZ antibody (1:300 dilution, BD Biosciences) showed that TAZ was overexpressed in the nuclei (iii). Pictures were taken using <t>TE200</t> Nikon Inverted Fluorescent Microscope (Nikon, Montreal, Canada) as 20× magnification. ( B ) Western blot analysis of TAZ-S89A expression. Ten μg of cell lysate extracted from E10-TAZS89A-P or E10-TAZ-S89A-T cells in the absence (−) or presence (+) of Dox were subjected to WB analysis using anti-TAZ (1:1000, BD Biosciences) and anti β-actin (1:10,000 Sigma, Oakville, Canada) antibodies. β-actin was used as an internal loading control. ( C ) Cell proliferation assay. Triplicates of 1.5 × 10 4 E10-TAZ-S89A-P or tumorigenic E10-TAZ-S89A-T cells were seeded into each well of 12-well plates, then untreated (−) or treated (+) with Dox. Cell numbers were counted on days 1, 2, 3, 4, 5, and 6 after plating. The experiments were repeated at least three times. Data are shown as means ± S.D. “*” represent significant difference. ( D – E ) Soft agar assay. Triplicates of 2×10 3 TAZ-S89A-P or E10-TAZ-S89A-T cells were mixed with 0.4% agarose in growth media and overlaid on 0.8% agarose in 6-well plates, followed by incubation in the absence or presence of Dox (2 μg/ml). Colony formation was examined after culturing for 18 days (D). Colony numbers were counted by Bio-Rad Gel Doc System (Bio-Rad, Mississauga, Canada). Data are shown as means ± S.D. “*” represent significant difference ( P
    Fluorescence Inverse Microscope Nikon Eclipse80i, supplied by Nikon, used in various techniques. Bioz Stars score: 87/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon fluorescent nikon ts 100 inverted microscope
    Establishment of an in vivo xenograft TAZ-overexpressing mouse model ( A ) Overexpression of TAZ-S89A in mouse immortalized lung epithelial cells (E10) caused highly malignant NSCLC tumor formation. Tumorigenesis assay was performed by subcutaneously injecting about 3 × 10 6 E10-TAZ-S89A cells into two-sides of nude mice. E10-TAZ-S89A cells caused large tumors (i) in two weeks. Two week later, the tumors were fixed, sectioned, and subjected to H E staining and IHC. H E staining with antibody incubation of E10-TAZ-S89A tumor section showed high-grade, poorly-differentiated carcinoma (ii). IHC staining for TAZ expression using TAZ antibody (1:300 dilution, BD Biosciences) showed that TAZ was overexpressed in the nuclei (iii). Pictures were taken using <t>TE200</t> Nikon Inverted Fluorescent Microscope (Nikon, Montreal, Canada) as 20× magnification. ( B ) Western blot analysis of TAZ-S89A expression. Ten μg of cell lysate extracted from E10-TAZS89A-P or E10-TAZ-S89A-T cells in the absence (−) or presence (+) of Dox were subjected to WB analysis using anti-TAZ (1:1000, BD Biosciences) and anti β-actin (1:10,000 Sigma, Oakville, Canada) antibodies. β-actin was used as an internal loading control. ( C ) Cell proliferation assay. Triplicates of 1.5 × 10 4 E10-TAZ-S89A-P or tumorigenic E10-TAZ-S89A-T cells were seeded into each well of 12-well plates, then untreated (−) or treated (+) with Dox. Cell numbers were counted on days 1, 2, 3, 4, 5, and 6 after plating. The experiments were repeated at least three times. Data are shown as means ± S.D. “*” represent significant difference. ( D – E ) Soft agar assay. Triplicates of 2×10 3 TAZ-S89A-P or E10-TAZ-S89A-T cells were mixed with 0.4% agarose in growth media and overlaid on 0.8% agarose in 6-well plates, followed by incubation in the absence or presence of Dox (2 μg/ml). Colony formation was examined after culturing for 18 days (D). Colony numbers were counted by Bio-Rad Gel Doc System (Bio-Rad, Mississauga, Canada). Data are shown as means ± S.D. “*” represent significant difference ( P
    Fluorescent Nikon Ts 100 Inverted Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Nikon inverted nikon ti e fluorescence microscope
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Inverted Nikon Ti E Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 79/100, based on 1059 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    82
    Nikon tie inverted widefield fluorescence nikon microscope
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Tie Inverted Widefield Fluorescence Nikon Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 82/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon te2000e nikon inverted epi fluorescence microscope
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Te2000e Nikon Inverted Epi Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Nikon fluorescence inverted nikon confocal microscope
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Fluorescence Inverted Nikon Confocal Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 90/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Nikon fluorescence microscopy images nikon tis inverted fluorescence microscopy
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Fluorescence Microscopy Images Nikon Tis Inverted Fluorescence Microscopy, supplied by Nikon, used in various techniques. Bioz Stars score: 77/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Nikon inverse nikon ti e fluorescence microscope
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Inverse Nikon Ti E Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 93/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Nikon inverse fluorescence microscope nikon diaphot tmd
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Inverse Fluorescence Microscope Nikon Diaphot Tmd, supplied by Nikon, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon custom automated nikon inverted fluorescence microscope
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Custom Automated Nikon Inverted Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 90/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon wide field inverted nikon te2000u fluorescence microscope
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Wide Field Inverted Nikon Te2000u Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 78/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon eclipse ts 100f nikon inverted fluorescent microscope
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Eclipse Ts 100f Nikon Inverted Fluorescent Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 84/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon inverted te300 nikon wide field fluorescence microscope
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Inverted Te300 Nikon Wide Field Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 78/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon widefield fluorescent nikon eclipse inverted microscope
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
    Widefield Fluorescent Nikon Eclipse Inverted Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nikon fluorescence imaging a nikon te2000 inverted microscope
    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
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    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon <t>Eclipse</t> <t>Ti</t> <t>inverted</t> <t>fluorescence</t> <t>microscope.</t> White bar indicates length of 100 μm.
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    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence <t>NIKON</t> eclipse <t>T2000-U</t> microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p
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    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence <t>NIKON</t> eclipse <t>T2000-U</t> microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p
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    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence <t>NIKON</t> eclipse <t>T2000-U</t> microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p
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    Nikon bright field nikon eclipse ts100 inverted fluorescent microscope
    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence <t>NIKON</t> eclipse <t>T2000-U</t> microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p
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    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence <t>NIKON</t> eclipse <t>T2000-U</t> microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p
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    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence <t>NIKON</t> eclipse <t>T2000-U</t> microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p
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    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence <t>NIKON</t> eclipse <t>T2000-U</t> microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p
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    Nikon laminar flow • cell counter chamber • nikon eclipse inverted fluorescence microscope
    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence <t>NIKON</t> eclipse <t>T2000-U</t> microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p
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    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence <t>NIKON</t> eclipse <t>T2000-U</t> microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p
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    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence <t>NIKON</t> eclipse <t>T2000-U</t> microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p
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    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence <t>NIKON</t> eclipse <t>T2000-U</t> microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p
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    Image Search Results


    Global nascent protein synthesis is repressed in Zscan4 + state cells and has elevated levels of Eif1a. (A) To determine global protein synthesis, mouse ES cells were cultured in methionine-free ES cell media for 30 min, labelled with AHA, a methionine analogue, for 1 h and visualized with Alexa Fluor 488 alkyne through Click-iT reaction and co-stained with Zscan4 antibody. Zscan + ES cells (i, arrowhead) showed dramatic repression in AHA staining (ii and iii). Eif1a levels were determined in Zscan4 + cells by co-staining with the respective antibodies. Zscan4 + cells (iv) showed enhanced Eif1a expression (v and vi). Pictures were taken at ×40 magnification using a Nikon Eclipse TE300 microscope (scale bar indicates 50 mm). (B) pZscan4 -puro mouse ES cells were treated with (+) or without (−) 1.5 mg/ml puromycin for 1 or 2 days. Cells were labelled with AHA as described above and protein harvested using RIPA buffer containing protease inhibitors. AHA-labelled samples were then detected using TAMRA alkyne via Click-iT reaction. Samples were run on a 4–12% Bis-Tris SDS–PAGE acrylamide gel, and TAMRA was visualized under UV illuminescence. (C) Western blot analysis was performed on the same gel (C) to determine the levels of Zscan4 and Eif1a. Protein from the acrylamide gel was transferred onto the polyvinylidene difluoride (PVDF) membrane and blotted with antibodies against Zscan4, Eif1a, and beta-actin.

    Journal: DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes

    Article Title: Repression of Global Protein Synthesis by Eif1a-Like Genes That Are Expressed Specifically in the Two-Cell Embryos and the Transient Zscan4-Positive State of Embryonic Stem Cells

    doi: 10.1093/dnares/dst018

    Figure Lengend Snippet: Global nascent protein synthesis is repressed in Zscan4 + state cells and has elevated levels of Eif1a. (A) To determine global protein synthesis, mouse ES cells were cultured in methionine-free ES cell media for 30 min, labelled with AHA, a methionine analogue, for 1 h and visualized with Alexa Fluor 488 alkyne through Click-iT reaction and co-stained with Zscan4 antibody. Zscan + ES cells (i, arrowhead) showed dramatic repression in AHA staining (ii and iii). Eif1a levels were determined in Zscan4 + cells by co-staining with the respective antibodies. Zscan4 + cells (iv) showed enhanced Eif1a expression (v and vi). Pictures were taken at ×40 magnification using a Nikon Eclipse TE300 microscope (scale bar indicates 50 mm). (B) pZscan4 -puro mouse ES cells were treated with (+) or without (−) 1.5 mg/ml puromycin for 1 or 2 days. Cells were labelled with AHA as described above and protein harvested using RIPA buffer containing protease inhibitors. AHA-labelled samples were then detected using TAMRA alkyne via Click-iT reaction. Samples were run on a 4–12% Bis-Tris SDS–PAGE acrylamide gel, and TAMRA was visualized under UV illuminescence. (C) Western blot analysis was performed on the same gel (C) to determine the levels of Zscan4 and Eif1a. Protein from the acrylamide gel was transferred onto the polyvinylidene difluoride (PVDF) membrane and blotted with antibodies against Zscan4, Eif1a, and beta-actin.

    Article Snippet: Fluorescence was visualized and captured using a Nikon Eclipse TE300 inverted microscope (Nikon, Melville, NY, USA).

    Techniques: Cell Culture, Staining, Expressing, Microscopy, SDS Page, Acrylamide Gel Assay, Western Blot

    Global nascent protein synthesis observed in Zscan4+ state cells not due to stress-induced response or cells being blocked in the mitosis stage of cell cycle. (A) Mouse ES cells were co-immunostained with antibodies against Zscan4 (i) and Eif2α phosphoserine 51 antibodies (ii) and visualized using Alexa Fluor 568 and 488, respectively. DNA was visualized by 4′,6-diamidino-2-phenylindole, dihydrochloride (DAPI) staining (iii). White arrows indicate the position of Zscan4 + state cells. Pictures were taken at ×40 magnification using a Nikon Eclipse TE300 microscope (scale bar indicates 50 mm). White arrows indicate Zscan4-positive cells. (B) To determine global protein synthesis, mouse ES cells were cultured in methionine-free ES cell media for 30 min, labelled with AHA, a methionine analogue, for 1 h and visualized with Alexa Fluor 488 alkyne through Click-iT reaction (i) and co-stained with histone H3 phosphoserine 10 (H3S10ph) antibody (ii). DNA was visualized by DAPI staining (iv and viii). White arrows indicate the position of H3S10ph stained cells in i–iv. To investigate whether cells under Zscan4 + state are blocked in the mitotic phase of the cell cycle, mouse ES cells were co-immunostained with Zscan4 (v) and H3S10ph (vi) antibodies. Pictures were taken at ×40 magnification using a Nikon Eclipse TE300 microscope (scale bar indicates 50 mm).

    Journal: DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes

    Article Title: Repression of Global Protein Synthesis by Eif1a-Like Genes That Are Expressed Specifically in the Two-Cell Embryos and the Transient Zscan4-Positive State of Embryonic Stem Cells

    doi: 10.1093/dnares/dst018

    Figure Lengend Snippet: Global nascent protein synthesis observed in Zscan4+ state cells not due to stress-induced response or cells being blocked in the mitosis stage of cell cycle. (A) Mouse ES cells were co-immunostained with antibodies against Zscan4 (i) and Eif2α phosphoserine 51 antibodies (ii) and visualized using Alexa Fluor 568 and 488, respectively. DNA was visualized by 4′,6-diamidino-2-phenylindole, dihydrochloride (DAPI) staining (iii). White arrows indicate the position of Zscan4 + state cells. Pictures were taken at ×40 magnification using a Nikon Eclipse TE300 microscope (scale bar indicates 50 mm). White arrows indicate Zscan4-positive cells. (B) To determine global protein synthesis, mouse ES cells were cultured in methionine-free ES cell media for 30 min, labelled with AHA, a methionine analogue, for 1 h and visualized with Alexa Fluor 488 alkyne through Click-iT reaction (i) and co-stained with histone H3 phosphoserine 10 (H3S10ph) antibody (ii). DNA was visualized by DAPI staining (iv and viii). White arrows indicate the position of H3S10ph stained cells in i–iv. To investigate whether cells under Zscan4 + state are blocked in the mitotic phase of the cell cycle, mouse ES cells were co-immunostained with Zscan4 (v) and H3S10ph (vi) antibodies. Pictures were taken at ×40 magnification using a Nikon Eclipse TE300 microscope (scale bar indicates 50 mm).

    Article Snippet: Fluorescence was visualized and captured using a Nikon Eclipse TE300 inverted microscope (Nikon, Melville, NY, USA).

    Techniques: Staining, Microscopy, Cell Culture

    Over-expression of Eif1a-like genes represses global nascent protein translation. (A) mESC were transiently transfected with FLAG-tagged Eif1a (i–iv) or FLAG- Eif1a -like genes, Eif1al6 (v–viii) or Eif1al9 (ix–xii), constructs for 18 h before selection with puromycin for 24 h. Cells were then cultured in methionine-free ES cell media for 30 min and labelled with AHA for 1 h. AHA-labelled cells were visualized with Alexa Fluor 488 alkyne through Click-iT reaction (ii, vi, and x) and co-stained with FLAG antibody (i, v, and ix). DNA was visualized by DAPI staining (iv, viii, and xii). Pictures were taken at ×40 magnification using a Nikon Eclipse TE300 microscope (scale bar indicates 50 mm). White arrows indicated cells with high levels of FLAG- Eif1a or FLAG- Eif1a - like genes over-expression. (B) Cells were labelled with AHA as described above and protein harvested using RIPA buffer containing protease inhibitors. AHA-labelled samples were then detected using TAMRA alkyne via Click-iT reaction (i). Samples were run on a 4–12% Bis-Tris SDS–PAGE acrylamide gel, transferred on the PVDF membrane, and blotted with antibodies against Zscan4, Eif1a and beta-actin (ii). (C) The model of global protein synthesis repression in Zscan4 + and Zscan4 − states in mouse ES cells. Diagram modified from Zalzman et al. 6

    Journal: DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes

    Article Title: Repression of Global Protein Synthesis by Eif1a-Like Genes That Are Expressed Specifically in the Two-Cell Embryos and the Transient Zscan4-Positive State of Embryonic Stem Cells

    doi: 10.1093/dnares/dst018

    Figure Lengend Snippet: Over-expression of Eif1a-like genes represses global nascent protein translation. (A) mESC were transiently transfected with FLAG-tagged Eif1a (i–iv) or FLAG- Eif1a -like genes, Eif1al6 (v–viii) or Eif1al9 (ix–xii), constructs for 18 h before selection with puromycin for 24 h. Cells were then cultured in methionine-free ES cell media for 30 min and labelled with AHA for 1 h. AHA-labelled cells were visualized with Alexa Fluor 488 alkyne through Click-iT reaction (ii, vi, and x) and co-stained with FLAG antibody (i, v, and ix). DNA was visualized by DAPI staining (iv, viii, and xii). Pictures were taken at ×40 magnification using a Nikon Eclipse TE300 microscope (scale bar indicates 50 mm). White arrows indicated cells with high levels of FLAG- Eif1a or FLAG- Eif1a - like genes over-expression. (B) Cells were labelled with AHA as described above and protein harvested using RIPA buffer containing protease inhibitors. AHA-labelled samples were then detected using TAMRA alkyne via Click-iT reaction (i). Samples were run on a 4–12% Bis-Tris SDS–PAGE acrylamide gel, transferred on the PVDF membrane, and blotted with antibodies against Zscan4, Eif1a and beta-actin (ii). (C) The model of global protein synthesis repression in Zscan4 + and Zscan4 − states in mouse ES cells. Diagram modified from Zalzman et al. 6

    Article Snippet: Fluorescence was visualized and captured using a Nikon Eclipse TE300 inverted microscope (Nikon, Melville, NY, USA).

    Techniques: Over Expression, Transfection, Construct, Selection, Cell Culture, Staining, Microscopy, SDS Page, Acrylamide Gel Assay, Modification

    TM effects insulin-stimulated Glut4 translocation and expression and glucose uptake in insulin-resistant C2C12 myotubes. A and B : TM enhanced insulin-stimulated Glut4 translocation and expression in palmitate-free (control [CON]) and palmitate-treated (PA) C2C12 myotubes, as shown by immunofluorescence, which was attenuated by PPARδ and PI3K inhibitors. The green fluorescence indicates Glut4. Nuclei in all groups were stained in blue with DAPI. The images were collected using a Nikon TE2000-U inverted fluorescence microscope ( A ) and total internal reflection microscopy (TIRFM) ( B ). Experiments were repeated three times. C : TM increased insulin-stimulated glucose uptake in palmitate-treated C2C12 myotubes. TM did not increase insulin-stimulated glucose uptake without palmitate exposure. The [ 3 H]-2-deoxyglucose uptake assays were performed in C2C12 myotubes. * P

    Journal: Diabetes

    Article Title: Telmisartan Improves Insulin Resistance of Skeletal Muscle Through Peroxisome Proliferator-Activated Receptor-? Activation

    doi: 10.2337/db12-0570

    Figure Lengend Snippet: TM effects insulin-stimulated Glut4 translocation and expression and glucose uptake in insulin-resistant C2C12 myotubes. A and B : TM enhanced insulin-stimulated Glut4 translocation and expression in palmitate-free (control [CON]) and palmitate-treated (PA) C2C12 myotubes, as shown by immunofluorescence, which was attenuated by PPARδ and PI3K inhibitors. The green fluorescence indicates Glut4. Nuclei in all groups were stained in blue with DAPI. The images were collected using a Nikon TE2000-U inverted fluorescence microscope ( A ) and total internal reflection microscopy (TIRFM) ( B ). Experiments were repeated three times. C : TM increased insulin-stimulated glucose uptake in palmitate-treated C2C12 myotubes. TM did not increase insulin-stimulated glucose uptake without palmitate exposure. The [ 3 H]-2-deoxyglucose uptake assays were performed in C2C12 myotubes. * P

    Article Snippet: The images were collected using a Nikon TE2000-U inverted fluorescence microscope (Nikon, Tokyo, Japan) and total internal reflection microscopy (Nikon) ( ) ( Supplementary Data ).

    Techniques: Translocation Assay, Expressing, Immunofluorescence, Fluorescence, Staining, Microscopy

    Phase contrast and fluorescence images showing the internalization and intracellular distribution of cationic G5-(Fl)-(NH 2 ) 122 , acetylated G5-(Fl)-(Ac) 110 , and G5-(Fl)-(Ac) 83 -(NAcGal) 14 conjugates into HepG2 cells visualized at a 20× magnification (scale bar = 100 μm) using a Nikon Eclipse TE2000-U inverted microscope with a Photometrics EMCCD camera and EXFO fluorescent lamp ( λ ex = 488 nm, λ em = 512 nm).

    Journal: Biomaterials

    Article Title: N-acetylgalactosamine-functionalized dendrimers as hepatic cancer cell-targeted carriers

    doi: 10.1016/j.biomaterials.2010.11.068

    Figure Lengend Snippet: Phase contrast and fluorescence images showing the internalization and intracellular distribution of cationic G5-(Fl)-(NH 2 ) 122 , acetylated G5-(Fl)-(Ac) 110 , and G5-(Fl)-(Ac) 83 -(NAcGal) 14 conjugates into HepG2 cells visualized at a 20× magnification (scale bar = 100 μm) using a Nikon Eclipse TE2000-U inverted microscope with a Photometrics EMCCD camera and EXFO fluorescent lamp ( λ ex = 488 nm, λ em = 512 nm).

    Article Snippet: Uptake of non-acetylated G5-(Fl)-(NH2 )122 , acetylated G5-(Fl)-(Ac)110 , and NAcGal-targeted G5-(Fl)-(Ac)83 -(NAcGal)14 conjugates into HepG2 cells (1 × 105 cells/well) upon incubation with 200 n m solution of each conjugate for 4 h was visualized using a Nikon Eclipse TE2000-U inverted microscope equipped with a photometrics EMCCD camera and an EXFO fluorescent lamp ( λ ex = 488 nm, λ em = 512 nm).

    Techniques: Fluorescence, Inverted Microscopy

    Cd-induced extracellular Ca 2 + influx elevates [Ca 2 + ] i contributing to neuronal apoptosis via activation of MAPK and mTOR pathways. Indicated cells were pretreated with or without 100 µM EGTA for 30 min, and then exposed to Cd (10 and/or 20 µM) for 24 h (A–D) or 4 h (E, F). (A and B) [Ca 2+ ] i fluorescent intensities were evaluated as described in Materials and Methods . (C) Morphology of PC12 cells was assessed using a Nikon Eclipse TE2000-U inverted phase-contrast microscope (200×) equipped with digital camera. (D) Cell viability in SH-SY5Y cells and primary neurons was evaluated by one solution assay. (E and F) Indicated cell lysates were subjected to Western blotting using indicated antibodies. The blots were probed for β-tubulin as a loading control. Similar results were observed in at least three independent experiments. Results (A, D) are presented as mean ± SE; n = 6. b P

    Journal: PLoS ONE

    Article Title: Calcium Signaling Is Involved in Cadmium-Induced Neuronal Apoptosis via Induction of Reactive Oxygen Species and Activation of MAPK/mTOR Network

    doi: 10.1371/journal.pone.0019052

    Figure Lengend Snippet: Cd-induced extracellular Ca 2 + influx elevates [Ca 2 + ] i contributing to neuronal apoptosis via activation of MAPK and mTOR pathways. Indicated cells were pretreated with or without 100 µM EGTA for 30 min, and then exposed to Cd (10 and/or 20 µM) for 24 h (A–D) or 4 h (E, F). (A and B) [Ca 2+ ] i fluorescent intensities were evaluated as described in Materials and Methods . (C) Morphology of PC12 cells was assessed using a Nikon Eclipse TE2000-U inverted phase-contrast microscope (200×) equipped with digital camera. (D) Cell viability in SH-SY5Y cells and primary neurons was evaluated by one solution assay. (E and F) Indicated cell lysates were subjected to Western blotting using indicated antibodies. The blots were probed for β-tubulin as a loading control. Similar results were observed in at least three independent experiments. Results (A, D) are presented as mean ± SE; n = 6. b P

    Article Snippet: Finally, calcium imaging was acquired with a Nikon Eclipse TE2000-U inverted fluorescence microscope (Melville, NY, USA) equipped with a digital camera.

    Techniques: Activation Assay, Microscopy, Western Blot

    Establishment of an in vivo xenograft TAZ-overexpressing mouse model ( A ) Overexpression of TAZ-S89A in mouse immortalized lung epithelial cells (E10) caused highly malignant NSCLC tumor formation. Tumorigenesis assay was performed by subcutaneously injecting about 3 × 10 6 E10-TAZ-S89A cells into two-sides of nude mice. E10-TAZ-S89A cells caused large tumors (i) in two weeks. Two week later, the tumors were fixed, sectioned, and subjected to H E staining and IHC. H E staining with antibody incubation of E10-TAZ-S89A tumor section showed high-grade, poorly-differentiated carcinoma (ii). IHC staining for TAZ expression using TAZ antibody (1:300 dilution, BD Biosciences) showed that TAZ was overexpressed in the nuclei (iii). Pictures were taken using TE200 Nikon Inverted Fluorescent Microscope (Nikon, Montreal, Canada) as 20× magnification. ( B ) Western blot analysis of TAZ-S89A expression. Ten μg of cell lysate extracted from E10-TAZS89A-P or E10-TAZ-S89A-T cells in the absence (−) or presence (+) of Dox were subjected to WB analysis using anti-TAZ (1:1000, BD Biosciences) and anti β-actin (1:10,000 Sigma, Oakville, Canada) antibodies. β-actin was used as an internal loading control. ( C ) Cell proliferation assay. Triplicates of 1.5 × 10 4 E10-TAZ-S89A-P or tumorigenic E10-TAZ-S89A-T cells were seeded into each well of 12-well plates, then untreated (−) or treated (+) with Dox. Cell numbers were counted on days 1, 2, 3, 4, 5, and 6 after plating. The experiments were repeated at least three times. Data are shown as means ± S.D. “*” represent significant difference. ( D – E ) Soft agar assay. Triplicates of 2×10 3 TAZ-S89A-P or E10-TAZ-S89A-T cells were mixed with 0.4% agarose in growth media and overlaid on 0.8% agarose in 6-well plates, followed by incubation in the absence or presence of Dox (2 μg/ml). Colony formation was examined after culturing for 18 days (D). Colony numbers were counted by Bio-Rad Gel Doc System (Bio-Rad, Mississauga, Canada). Data are shown as means ± S.D. “*” represent significant difference ( P

    Journal: Oncotarget

    Article Title: TAZ induces lung cancer stem cell properties and tumorigenesis by up-regulating ALDH1A1

    doi: 10.18632/oncotarget.16430

    Figure Lengend Snippet: Establishment of an in vivo xenograft TAZ-overexpressing mouse model ( A ) Overexpression of TAZ-S89A in mouse immortalized lung epithelial cells (E10) caused highly malignant NSCLC tumor formation. Tumorigenesis assay was performed by subcutaneously injecting about 3 × 10 6 E10-TAZ-S89A cells into two-sides of nude mice. E10-TAZ-S89A cells caused large tumors (i) in two weeks. Two week later, the tumors were fixed, sectioned, and subjected to H E staining and IHC. H E staining with antibody incubation of E10-TAZ-S89A tumor section showed high-grade, poorly-differentiated carcinoma (ii). IHC staining for TAZ expression using TAZ antibody (1:300 dilution, BD Biosciences) showed that TAZ was overexpressed in the nuclei (iii). Pictures were taken using TE200 Nikon Inverted Fluorescent Microscope (Nikon, Montreal, Canada) as 20× magnification. ( B ) Western blot analysis of TAZ-S89A expression. Ten μg of cell lysate extracted from E10-TAZS89A-P or E10-TAZ-S89A-T cells in the absence (−) or presence (+) of Dox were subjected to WB analysis using anti-TAZ (1:1000, BD Biosciences) and anti β-actin (1:10,000 Sigma, Oakville, Canada) antibodies. β-actin was used as an internal loading control. ( C ) Cell proliferation assay. Triplicates of 1.5 × 10 4 E10-TAZ-S89A-P or tumorigenic E10-TAZ-S89A-T cells were seeded into each well of 12-well plates, then untreated (−) or treated (+) with Dox. Cell numbers were counted on days 1, 2, 3, 4, 5, and 6 after plating. The experiments were repeated at least three times. Data are shown as means ± S.D. “*” represent significant difference. ( D – E ) Soft agar assay. Triplicates of 2×10 3 TAZ-S89A-P or E10-TAZ-S89A-T cells were mixed with 0.4% agarose in growth media and overlaid on 0.8% agarose in 6-well plates, followed by incubation in the absence or presence of Dox (2 μg/ml). Colony formation was examined after culturing for 18 days (D). Colony numbers were counted by Bio-Rad Gel Doc System (Bio-Rad, Mississauga, Canada). Data are shown as means ± S.D. “*” represent significant difference ( P

    Article Snippet: Pictures were taken using TE200 Nikon Inverted Fluorescent Microscope at 40× magnification.

    Techniques: In Vivo, Over Expression, Mouse Assay, Staining, Immunohistochemistry, Incubation, Expressing, Microscopy, Western Blot, Proliferation Assay, Soft Agar Assay

    Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon Eclipse Ti inverted fluorescence microscope. White bar indicates length of 100 μm.

    Journal: Antiviral research

    Article Title: In vitro antiviral activity of adenosine analog NITD008 against tick-borne flaviviruses

    doi: 10.1016/j.antiviral.2016.03.013

    Figure Lengend Snippet: Immunofluorescence Assay (IFA). A549 cells were pretreated with NITD008, infected with indicated viruses for 24 h. Cells were fixed for 15 min in 10% formalin supplemented with 0.2% Triton-X detergent, then stained with a primary anti-flavivirus HMAF and a secondary donkey anti-mouse antibody conjugated with DyLight 488 (green). Cells were counterstained with DAPI (blue), and images were captured at 40 × magnification using a Nikon Eclipse Ti inverted fluorescence microscope. White bar indicates length of 100 μm.

    Article Snippet: Cells were counterstained with 4′,6-diamidino-2-phenylindole (DAPI) (Invitrogen, Carlsbad, CA USA), and images were captured at 40 × magnification using a Nikon Eclipse Ti inverted fluorescence microscope (Nikon, Melville, NY, USA).

    Techniques: Immunofluorescence, Infection, Staining, Fluorescence, Microscopy

    Distinct morphological changes in hMSCs co-cultured with cancer cell lines. a – l GFP-labeled hMSCs were co-cultured with the indicated tumor cell lines at a 1:2 ratio, and on day 7 of the co-culture images were taken using a Nikon® ECLIPSE Ti-U inverted fluorescence microscope (4×). Data are representative of at least three independent experiments. m hMSCs co-cultured with normal breast epithelial cell line (MCF10A). n hMSCs (GFP-labeled) co-cultured with MCF7 (mcherry-labeled) as above. Cells were stained with nuclear staining DAPI ( blue ). o AT-MSCs labeled with DiO ( green ) and co-cultured with MCF7 cells labeled with DiD ( red ). At day 7, photomicrographs were taken using a Nikon® ECLIPSE Ti-U inverted fluorescence microscope (magnification 4×). p MCF7 cells were co-cultured with hMSCs at the indicated ratios (tumor:MSCs), and on day 7 images were captured in the green channel (MSCs) using 4× magnification. DAPI was used to stain nuclei

    Journal: Stem Cell Research & Therapy

    Article Title: CDH1 and IL1-beta expression dictates FAK and MAPKK-dependent cross-talk between cancer cells and human mesenchymal stem cells

    doi: 10.1186/s13287-015-0123-0

    Figure Lengend Snippet: Distinct morphological changes in hMSCs co-cultured with cancer cell lines. a – l GFP-labeled hMSCs were co-cultured with the indicated tumor cell lines at a 1:2 ratio, and on day 7 of the co-culture images were taken using a Nikon® ECLIPSE Ti-U inverted fluorescence microscope (4×). Data are representative of at least three independent experiments. m hMSCs co-cultured with normal breast epithelial cell line (MCF10A). n hMSCs (GFP-labeled) co-cultured with MCF7 (mcherry-labeled) as above. Cells were stained with nuclear staining DAPI ( blue ). o AT-MSCs labeled with DiO ( green ) and co-cultured with MCF7 cells labeled with DiD ( red ). At day 7, photomicrographs were taken using a Nikon® ECLIPSE Ti-U inverted fluorescence microscope (magnification 4×). p MCF7 cells were co-cultured with hMSCs at the indicated ratios (tumor:MSCs), and on day 7 images were captured in the green channel (MSCs) using 4× magnification. DAPI was used to stain nuclei

    Article Snippet: The scratch area was imaged on day 0 and day 2 using 4× magnification with the Nikon® ECLIPSE Ti-U inverted fluorescence microscope.

    Techniques: Cell Culture, Labeling, Co-Culture Assay, Fluorescence, Microscopy, Staining

    Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence NIKON eclipse T2000-U microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p

    Journal: ASN NEURO

    Article Title: Both Creatine and Its Product Phosphocreatine Reduce Oxidative Stress and Afford Neuroprotection in an In Vitro Parkinson’s Model

    doi: 10.1177/1759091414554945

    Figure Lengend Snippet: Creatine and PCr inhibit oxidative stress induced by 6-OHDA in rat striatal slices. ROS generation was estimated with the fluorescent probe, 2′,7′-dichlorofluorescein diacetate (H 2 DCFDA). DCF fluorescence images in the putamen (a–d) and globus pallidus (e–h) under the different experimental conditions indicated in each panel taken at 100×. Quantification of DCF fluorescence on rat striatal slices homogenates was measured in a Tecan microplate reader and expressed as nmol of oxidized DCF per mg protein and normalized to control 100%. The quantification of DCF fluorescence in the putamen (j) and globus pallidus (k) was measured in an epifluorescence NIKON eclipse T2000-U microscope. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the mean + SEM of six to nine experiments. ** p

    Article Snippet: Fluorescence was measured in a fluorescence-inverted NIKON eclipse T2000-U microscope (Nikon Instruments Europe, Badhoevedorp, the Netherlands).

    Techniques: Polymerase Chain Reaction, Fluorescence, Microscopy

    Creatine or PCr prevent loss of mitochondrial membrane potential in the rat striatal slices subjected to 6-OHDA. TMRE fluorescence in the putamen (a–d) and globus pallidus (e–h) was analyzed 4 h after coincubation with 6-OHDA plus creatine or PCr, as detailed in the Materials and Methods section. The quantification of TMRE fluorescence was measured in an epifluorescence NIKON eclipse T2000-U microscope (i, j). The top part of the figure illustrates representative photomicrographs of putamen and globus pallidus at 100×. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the means + SEM of six to eight experiments carried out in triplicates. ** p

    Journal: ASN NEURO

    Article Title: Both Creatine and Its Product Phosphocreatine Reduce Oxidative Stress and Afford Neuroprotection in an In Vitro Parkinson’s Model

    doi: 10.1177/1759091414554945

    Figure Lengend Snippet: Creatine or PCr prevent loss of mitochondrial membrane potential in the rat striatal slices subjected to 6-OHDA. TMRE fluorescence in the putamen (a–d) and globus pallidus (e–h) was analyzed 4 h after coincubation with 6-OHDA plus creatine or PCr, as detailed in the Materials and Methods section. The quantification of TMRE fluorescence was measured in an epifluorescence NIKON eclipse T2000-U microscope (i, j). The top part of the figure illustrates representative photomicrographs of putamen and globus pallidus at 100×. Quantification of the mean fluorescence obtained under each experimental condition in putamen and globus pallidus is normalized respect to control (100%). Each column represents the means + SEM of six to eight experiments carried out in triplicates. ** p

    Article Snippet: Fluorescence was measured in a fluorescence-inverted NIKON eclipse T2000-U microscope (Nikon Instruments Europe, Badhoevedorp, the Netherlands).

    Techniques: Polymerase Chain Reaction, Fluorescence, Microscopy