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Fv 1000 filter confocal system

Manufactured by Olympus
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The FV 1000 Filter Confocal system is a high-performance confocal microscope designed for advanced imaging applications. It features a modular design that allows for customization to fit specific research needs. The system utilizes a laser-scanning technology to provide high-resolution imaging of various biological samples.

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Lab products found in correlation

Ds qi2 camera, by National Instruments (2 mentions) Eclipse ni u microscope, by National Instruments (2 mentions) Imagej, by Olympus (1 mentions) Fluoroview software, by Olympus (1 mentions) Fv10 asw software, by Olympus (1 mentions) Eclipse ni u microscope, by Nikon (1 mentions) Ds ri2, by National Instruments (1 mentions) Ds qi2, by National Instruments (1 mentions) Ultra 55, by Zeiss (1 mentions) Ti s epifluorescence microscope, by Nikon (1 mentions) Plan infinity corrected objective, by Mitutoyo (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) Calcien am, by Thermo Fisher Scientific (1 mentions)

8 protocols using fv 1000 filter confocal system

1

Immunofluorescence Localization of Exogenous FOXA2

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Transfected cells plated on coverslips were fixed and permeabilized using standard methods. Exogenous FOXA2 was detected using the 9E10 anti-myc antibody. DAPI was used to stain nuclei. Imaging was performed using a Keyence BZ-9000 florescent microscope (Keyence Corporation, Itasca, IL, USA). An Olympus FV 1000 Filter Confocal system (Olympus America Inc, Melville, NY, USA) was used for confocal imaging.
Neff R., Rush C.M., Smith B., Backes F.J., Cohn D.E, & Goodfellow P.J. (2018). Functional characterization of recurrent FOXA2 mutations seen in endometrial cancers. International journal of cancer, 143(11), 2955-2961.
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2

Immunofluorescence Imaging of Cell Nuclei

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Cells were grown on glass coverslips, fixed in 4% paraformaldehyde, and permeabilized in 0.1% Tween20. Coverslips were blocked in 5% normal goat serum for 1 hour and incubated with primary antibody overnight in a humidified chamber at 4°C. They were then washed with PBS and incubated with Alexaflour 488 secondary antibody for 1 hour. Coverslips were counterstained with DAPI and mounted on glass slides with Vectashield Hardset. All images were taken on an Olympus FV 1000 Filter Confocal System with a super-corrected 60x oil objective. Image adjustment and analysis was performed in Olympus Fluoroview software and ImageJ. Z-stacks were generated with summed intensities and an image mask was generated using an established algorithm (34 ). Median pixel intensity measured for each cell nucleus in the field; values from partial nuclei containing less than 60μm of area were removed. Data was then exported to R for statistical analysis.
Koenig M.J., Agana B.A., Kaufman J.M., Sharpnack M.F., Wang W.Z., Weigel C., Navarro F.C., Amann J.M., Cacciato N., Arasada R.R., Gerstein M.B., Wysocki V.H., Oakes C, & Carbone D.P. (2021). STK11/LKB1 loss of function is associated with global DNA hypomethylation and S-adenosyl-methionine depletion in human lung adenocarcinoma. Cancer research, 81(16), 4194-4204.
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3

Multichannel Fluorescent Imaging Workflow

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Wide-field micrographs were collected using a Nikon Eclipse Ni-U microscope with a DS-Ri2 (immunohistochemistry) or a DS-Qi2 (immunofluorescence) camera and NIS-Elements Advanced Research software. Confocal micrographs were collected using the Olympus FV 1000 Filter Confocal system in Campus Microscopy & Imaging Facility (CMIF) at the Ohio State University. Images were processed for presentation using Olympus FV10-ASW software (Ver. 4.02). For visualization purposes only, background auto-fluorescence and non-specific staining were corrected by adjusting the intensity range using Look-Up Tables (non-destructive modifications) so to primarily display pixels representing specific signal. Tissues of E2f3a−/− and wild-type mice were used to aid in these corrections in the case of E2F3A and MYC-tag immunostaining, respectively. ImageJ (https://imagej.net/Welcome) was used for manual quantification of immunostaining on multichannel fluorescent images.
Cuitiño M.C., Pécot T., Sun D., Kladney R., Okano-Uchida T., Shinde N., Saeed R., Perez-Castro A.J., Webb A., Liu T., Bae S.I., Clijsters L., Selner N., Coppola V., Timmers C., Ostrowski M.C., Pagano M, & Leone G. (2019). Two Distinct E2F Transcriptional Modules Drive Cell Cycles and Differentiation. Cell reports, 27(12), 3547-3560.e5.
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4

Widefield and Confocal Microscopy Techniques

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Widefield micrographs were collected using a Nikon Eclipse Ni-U microscope with a DS-Qi2 camera and NIS-Elements Advanced Research software. Confocal micrographs were collected using the Olympus FV 1000 Filter Confocal system in the Campus Microscopy and Imaging Facility at the Ohio State University.
Pécot T., Cuitiño M.C., Johnson R.H., Timmers C, & Leone G. (2022). Deep learning tools and modeling to estimate the temporal expression of cell cycle proteins from 2D still images. PLoS Computational Biology, 18(3), e1009949.
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5

Characterization of Fluorescent Nanodiamonds

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Samples were prepared by depositing 0.0005% (w/w) FNDs on a coverslip and drying in a vacuum oven for 2 hours. Scanning electron microscopy (SEM) images were acquired on a field emission scanning electron microscope (Zeiss Ultra 55) operating at 5 kV and a working distance of 7.9 mm. Size characterization was performed from the SEM images (n = 168) using Image J. The fluorescence emission spectrum was obtained using a Horiba ARAMIS Raman upright microscope (50× objective) and a 532 nm excitation laser. Confocal microscopy images were collected on an Olympus FV1000-Filter Confocal System using a 543 nm excitation laser and corresponding 655/55 nm barrier filters.
Suarez-Kelly L.P., Campbell A.R., Rampersaud I.V., Bumb A., Wang M.S., Butchar J.P., Tridandapani S., Yu L., Rampersaud A.A, & Carson WE I.I.I. (2016). Fluorescent nanodiamonds engage innate immune effector cells: a potential vehicle for targeted anti-tumor immunotherapy. Nanomedicine : nanotechnology, biology, and medicine, 13(3), 909-920.
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6

Fluorescence Characterization of Fluorescent Nanodiamonds

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Fluorescence emission spectra of FND were collected on a custom-built confocal spectrometer, using a 532 nm laser diode as the light source. The excitation laser was focused through a long working distance NIR microscope (100X objective) with NA = 0.7 (Mitutoyo Plan Infinity Corrected Objective). The fluorescence from the FND was filtered from the reflected laser light using a 532 nm notch filter, then passed through a grating, collected on a CCD camera, and analyzed with customized spectral software. An Olympus FV1000-Filter Confocal System was used to collect microscopy images. The excitation laser wavelength was 543 nm and that for emission detection was 655–755 nm. Images were also collected on a Nikon Ti–S epifluorescence microscope with a Texas Red filter cube and analyzed with NIS Elements imaging software. The size of FND was determined by DLS using a Brookhaven 90Plus nanoparticle size analyzer (BIC, Holtsville, NY). From SEM images (Figure 1b), the diamonds could be characterized as having an irregular, blocky appearance.
Suarez-Kelly L.P., Sun S.H., Ren C., Rampersaud I.V., Albertson D., Duggan M.C., Noel T.C., Courtney N., Buteyn N.J., Moritz C., Yu L., Yildiz V.O., Butchar J.P., Tridandapani S., Rampersaud A.A, & Carson WE I.I.I. (2021). Antibody Conjugation of Fluorescent Nanodiamonds for Targeted Innate Immune Cell Activation. ACS Applied Nano Materials, 4(3), 3122-3139.
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7

Widefield and Confocal Microscopy Protocol

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Widefield micrographs were collected using a Nikon Eclipse Ni-U microscope with a DS-Qi2 camera and NIS-Elements Advanced Research software. Confocal micrographs were collected using the Olympus FV 1000 Filter Confocal system in the Campus Microscopy and Imaging Facility at the Ohio State University.
Pécot T., Cuitiño M.C., Johnson R.H., Timmers C., & Leone G. (2021). Deep learning tools and modeling to estimate the temporal expression of cell cycle proteins from 2D still images.
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8

Vacuum-Hydrodynamic Cell Trapping Platform

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The microfabrication of the 3D-MEP platform is described in Figure S7. A vacuum-hydrodynamic method was adopted for cell trapping on microchannel array. The vacuum was generated from a pump controlled by a valve with a gauge read-out from 1 to 11 psi. The cell suspension was loaded into the reservoir, and a negative pressure was applied as the vacuum was turned on. By additional hydrodynamic force, the buffer flowed into the waste bottle, while the cells were led to the bottom and sat on the microchannels. A gentle pressure of vacuum force (6 psi = 0.41 barye) was used for cell trapping. To increase the trapping efficiency, the trapping procedure was repeated three times on the same chip. To test the trapping efficiency, cells were stained with Hoechst and visualized with a fluorescence microscope (Nikon Eclipse Ti. Nikon, Japan). The interaction between cell and trapping microchannel was further studied using confocal fluorescence microscopy. To reveal the trapped cells, K562 was labeled with Hoechst for nuclei and Calcien-AM for cytosol (Thermofisher Scientific). The whole chip was fixed with 4% paraformaldehyde and observed with the Olympus FV 1000 Filter Confocal system.
Chiang C.L., Hu E.Y., Chang L., Labanowska J., Zapolnik K., Mo X., Shi J., Doong T.J., Lozanski A., Yan P.S., Bundschuh R., Walker L.A., Gallego-Perez D., Lu W., Long M., Kim S., Heerema N.A., Lozanski G., Woyach J.A., Byrd J.C., Lee L.J, & Muthusamy N. (2022). Leukemia-initiating HSCs in chronic lymphocytic leukemia reveal clonal leukemogenesis and differential drug sensitivity. Cell reports, 40(3).
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