Fluoview 1000 confocal scanning system

Manufactured by Olympus

Sourced in United States, Canada

The Fluoview-1000 is a confocal scanning system designed for high-resolution fluorescence imaging. It features a compact, modular design and a range of compatible objectives to enable precise, detailed observation of biological samples.

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Lab products found in correlation

Vectorshield mounting medium with dapi, by Vector Laboratories (5 mentions) Nestin, by Thermo Fisher Scientific (2 mentions) Alexa fluor 594 conjugated goat anti rabbit, by Thermo Fisher Scientific (2 mentions) C myc, by Thermo Fisher Scientific (2 mentions) Oct3 4, by Thermo Fisher Scientific (2 mentions) Ssea 1, by Thermo Fisher Scientific (2 mentions) Mklf4, by Thermo Fisher Scientific (2 mentions) Alexa fluor 594 conjugated goat anti rabbit, by Abcam (2 mentions) Alexa fluor 594 conjugated donkey anti goat, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Fluoview 1000 (658 citations) Fluoview 1000 System (10 citations) FluoView 1000 confocal laser scanning system (8 citations) FluoView FV-1000 confocal laser scanning system (6 citations) Fluoview 1000 confocal system (5 citations)

5 protocols using fluoview 1000 confocal scanning system

Immunofluorescence Analysis of Tumor Spheroids

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Mary-X spheroids and Karen-X loose aggregates were subjected to single label immunofluorescence studies using Alexa Fluor 488-conjugated 24E10 (#3199) (Cell Signaling Technology, Inc.) which recognized both E-cad/FL as well as E-cad/NTF1. In order to immobilize the spheroids and loose aggregates, glass-bottom dishes were coated with Cell-TEK adhesive. The adherent spheroids were then fixed with 4% paraformaldehyde, after permeabilizing with TX-100 and blocking with normal goat serum. After washing with PBS 4–5 times, each for 10 min, the spheroids and aggregates were incubated with 24E10. The dishes were finally mounted with Vectorshield mounting medium with DAPI (#H-1200) (Vector Laboratories) and viewed with a Olympus Fluoview-1000 confocal scanning system.
Mary-X and Karen-X were also subjected to double label immunofluorescence studies. Double label immunofluorescence experiments were carried out using the following combinations of antibodies: Alexa Fluor 488-conjugated 24E10 which recognized both E-cad/NTF1 as well as E-cad/FL and goat polyclonal antibody to mouse podoplanin or CD31 (R&D Systems, Inc.) which recognized murine lymphatics or blood vessels followed by Alexa Fluor 594-conjugated donkey anti-goat (#A11058) (Invitrogen, Inc.). Tumor emboli were recognized by E-cadherin positivity surrounded by circumferential podoplanin (lymphatics) or CD31 positivity.

Modi A.P., Nguyen J.P., Wang J., Ahn J.S., Libling W.A., Klein J.M., Mazumder P, & Barsky S.H. (2022). Geometric tumor embolic budding characterizes inflammatory breast cancer. Breast Cancer Research and Treatment, 197(3), 461-478.

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iPSC Characterization by Immunofluorescence

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The iPSC clones were confirmed as IPSCs with a battery of rabbit anti-mouse IPSC markers including Oct3/4, Sox2, c-Myc, mKlf4, Nestin and SSEA-1 (Thermo Fisher Scientific, Waltham, MA, USA). The secondary antibody was an Alexa Fluor 594-conjugated goat anti-rabbit (Thermo Fisher Scientific), all used in accordance with the manufacturer’s conditions. To immobilize the iPSC clones, glass-bottom dishes were coated with Cell-TEK adhesive. The adherent iPSC was then fixed with 4% paraformaldehyde, after permeabilizing with TX-100 and blocking with normal goat serum. The iPSC clones were then incubated with the primary antibodies, washed, and incubated with the secondary antibodies. The dishes were finally mounted with Vectorshield mounting medium with DAPI (#H-1200) (Vector Laboratories, Burlingame, CA, USA) and viewed with an Olympus Fluoview-1000 confocal scanning system under different wavelengths.

Petrova S.C., Ahmad I., Nguyen C., Ferrell SD J.r., Wilhelm S.R., Ye Y, & Barsky S.H. (2020). Regulation of breast cancer oncogenesis by the cell of origin’s differentiation state. Oncotarget, 11(43), 3832-3848.

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Multilineage Differentiation Analysis

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For each of the lineages, proof of successful differentiation was obtained by the detection of specific biomarkers for each lineage: CD31 for endothelial, albumin for hepatic and osteocalcin for osteogenic. Cultures of the differentiated cells that grew as monolayers were subjected to double immunofluorescent studies using the following combinations of antibodies: rabbit anti-mouse CD31, rabbit anti-mouse albumin, rabbit anti-mouse osteocalcin followed by Alexa Fluor 594-conjugated goat anti-rabbit (all antibodies from Abcam, Cambridge, MA, USA). The adherent monolayers were then fixed with 4% paraformaldehyde, after permeabilization with TX-100 and blocking with normal goat serum. The spheroids were then incubated with the respective primary antibodies according to the manufacturer’s specifications, washed with PBS 4–5 times, and incubated with the secondary antibody again according to the manufacturer’s specifications. The dishes were finally mounted with Vectorshield mounting medium with DAPI (#H-1200) (Vector Laboratories) and viewed with an Olympus Fluoview-1000 confocal scanning system.

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Characterization of Mouse iPSC Clones

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The iPSC clones were confirmed as IPSCs with a battery of rabbit anti-mouse IPSC markers including Oct3/4, Sox2, c-Myc, mKlf4, nestin, and SSEA-1 (Thermo Fisher Scientific, Waltham, MA) The secondary antibody was an Alexa Fluor 594-conjugated goat anti-rabbit (Thermo Fisher Scientific), all used with the manufacturer’s conditions. In order to immobilize the iPSC clones, glass-bottom dishes were coated with Cell-TEK adhesive. The adherent iPSC cells were then fixed with 4% paraformaldehyde, after permeabilizing with TX-100 and blocking with normal goat serum. The iPSC clones were then incubated with the primary antibodies, washed, and followed by the secondary antibodies. The dishes were finally mounted with Vectorshield mounting medium with DAPI (#H-1200) (Vector Laboratories, Burlingame, CA) and viewed with an Olympus Fluoview-1000 confocal scanning system under different wave lengths.

Nguyen C., Nguyen J.P., Modi A.P., Ahmad I., Petrova S.C., Ferrell SD J.r., Wilhelm S.R., Ye Y., Schaue D, & Barsky S.H. (2021). Use of constitutive and inducible oncogene-containing iPSCs as surrogates for transgenic mice to study breast oncogenesis. Stem Cell Research & Therapy, 12, 301.

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Multilineage Differentiation Validation

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For each of the lineages, proof of successful differentiation was obtained by the detection of specific biomarkers for each lineage: CD31 for endothelial, albumin for hepatic, and osteocalcin for osteogenic. Cultures of the differentiated cells which grew as monolayers were subjected to double immunofluorescent studies using the following combinations of antibodies: rabbit anti-mouse CD31, rabbit anti-mouse albumin, rabbit anti-mouse osteocalcin followed by Alexa Fluor 594-conjugated goat anti-rabbit (all antibodies from Abcam, Cambridge, MA) The adherent monolayers were then fixed with 4% paraformaldehyde, after permeabilizing with TX-100 and blocking with normal goat serum. The spheroids were then incubated with the respective primary antibodies according to the manufacturer’s specifications, followed by washing with PBS 4–5 times and then followed by the secondary antibody again according to the manufacturer’s specifications. The dishes were finally mounted with Vectorshield mounting medium with DAPI (#H-1200) (Vector Laboratories) and viewed with an Olympus Fluoview-1000 confocal scanning system. Confirmatory studies using the same lineage-specific antibodies were carried out by Western blot.

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