Metafluor imaging software

Manufactured by Universal Imaging

Sourced in Canada, United States

Metafluor is an imaging software designed for fluorescence microscopy. It provides tools for image acquisition, processing, and analysis. The software supports a range of camera and microscope configurations, enabling researchers to capture and analyze fluorescent signals in their experiments.

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Lab products found in correlation

Sutter dg 4 175w xenon arc lamp, by Sutter Instruments (2 mentions) Axiovert 100 tv, by Zeiss (1 mentions) Micromax, by Teledyne (1 mentions) Cell tak, by BD (1 mentions) Lambda 10 2, by Sutter Instruments (1 mentions) Rc 42lp, by Warner Instruments (1 mentions) 175 w xenon arc lamp, by Sutter Instruments (1 mentions) Ix71 inverted microscope, by Olympus (1 mentions) Fura 2 am, by Merck Group (1 mentions) Ccd camera, by Olympus (1 mentions)

6 protocols using metafluor imaging software

Mitochondrial Membrane Potential Imaging

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Muscle fibers were isolated from FDB muscle and mitochondrial membrane potential measured by epifluorescence microscopy on the basis of the accumulation of TMRM fluorescence, as previously described (Lo Verso et al., 2014 (link)). Fibers were considered as depolarizing when they lost more than 10% of the initial value of TMRM fluorescence. Imaging was performed with a Zeiss Axiovert 100 TV inverted microscope equipped with a 12-bit digital cooled charge-coupled device camera (Micromax, Princeton Instruments). The results were analyzed with MetaFluor imaging software (Universal Imaging).

Mansueto G., Armani A., Viscomi C., D’Orsi L., De Cegli R., Polishchuk E.V., Lamperti C., Di Meo I., Romanello V., Marchet S., Saha P.K., Zong H., Blaauw B., Solagna F., Tezze C., Grumati P., Bonaldo P., Pessin J.E., Zeviani M., Sandri M, & Ballabio A. (2017). Transcription Factor EB Controls Metabolic Flexibility during Exercise. Cell Metabolism, 25(1), 182-196.

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Fluorescence Imaging of Rat mTAL

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Rats were anesthetized with isoflurane (2–5%) and isolation of mTAL tissue strips performed as described previously^{31 (link)}. Thin tissue strips containing mTALs were placed on a glass coverslip coated with the tissue adhesive Cell-Tak (BD Biosciences) for fluorescence imaging. Tissue strips containing mTALs were loaded with either dihydroethidium DHE (50 mmol/L) or 2',7'-Bis-(2-Carboxyethyl) -5-(And-6) - carboxyfluorescein (BCECF; 6 µmol/L) in HBSS for 1 hour at room temperature. Coverslips were placed on a heated imaging chamber maintained at 37°C (Warner Instruments) that allowed the rapid exchange of superfusion buffer and mounted on the stage of an inverted microscope (IX81 Olympus). mTAL were visualized with a ×40 water immersion objective lens. The signal was detected using a high-resolution digital camera (Photometrics Evolve, Roper Scientific). Excitation was provided by a Sutter DG-4 175W xenon arc lamp (Sutter Instruments) that allowed high-speed excitation wavelength switching. 3 to 10 mTAL epithelial cells were selected within each tissue strip to quantify changes in fluorescent intensity of dyes using Metafluor imaging software (Universal Imaging). Intracellular [Na⁺] was set at either 5 or 30mM using the same solutions as that used to set [Na⁺]_I in MØ.

Jin C., Sun J., Stilphen C.A., Smith S.M., Ocasio H., Bermingham B., Darji S., Guha A., Patel R., Geurts A.M., Jacob H.J., Lambert N.A, & O’Connor P.M. (2014). HV1 ACTS AS A SODIUM SENSOR AND PROMOTES SUPEROXIDE PRODUCTION IN MEDULLARY THICK ASCENDING LIMB OF DAHL SALT-SENSITIVE RATS. Hypertension, 64(3), 541-550.

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Fluorescence Imaging of Mesangial Cells

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Fluorescence measurements were obtained using IX71 inverted microscope (Olympus, Tokyo, Japan) attached with a 60× (numerical aperture 0.9) water-immersion objective lens as previously reported^{14 (link)}. The signal was detected by a cool CCD video camera Ixon (Andor Co., Tokyo, Japan) coupled to Lambda-10.2 (Sutter Instruments, CA), and excitation was provided by Sutter DG-4 and 175-W Xenon-Arc lamp (Sutter Instruments, Navato, CA). For the experiments, cover slips were placed in an imaging chamber (RC-42LP and RC-43C; Warner Instruments, Hamden, CT) mounted on the stage of the inverted microscope and maintained at 37 °C with TC-344B thermo-warmer for chamber and TA-29 thermo-warmer for perfusate (Warner Instruments, Hamden, CT). The changes in fluorescence intensity were quantified using MetaFluor imaging software (Universal Imaging Co., Ypsilanti, MI) for each experiment. To exclude the possibility of selection bias of the measured MCs, the fluorescence intensity of all living mesangial cells in each cover glass was measured.

Akaishi T., Abe M., Okuda H., Ishizawa K., Abe T., Ishii T, & Ito S. (2019). High glucose level and angiotensin II type 1 receptor stimulation synergistically amplify oxidative stress in renal mesangial cells. Scientific Reports, 9, 5214.

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Measuring Astrocyte Nitric Oxide Levels

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Astrocytes plated on glass coverslips were loaded with 5 µM DAF-FM in DMEM without serum at 37 °C for 45 min and then washed three times in Locke’s solution in mM: 154 NaCl, 5.4 KCl, 2.3 CaCl₂, 5 HEPES, pH 7.4; followed by de-esterification at 37 °C for 15 min. Coverslips were then mounted in an Olympus BX 51W1I upright microscope with a 40x water immersion objective. Pictures were captured at 495 excitation wavelength and the emission was filtered at 505–550 nm using a Retiga 1300I fast-cooling monochrome digital monochrome camera (12-bit; Q Imaging, Burnaby, BC, Canada) controlled by Metafluor imaging software (Universal Imaging, Downingtown, PA, USA). Fluorescence intensity captured from at least 25 regions of interest (representing 25 cells per cultured coverslip) was calculated with the following formula: Corrected total cell fluorescence = Integrated Density – ([Area of selected cell] x [Mean fluorescence of background readings]).

Gómez G.I., Alvear T.F., Roa D.A., Farias-Pasten A., Vergara S.A., Mellado L.A., Martinez-Araya C.J., Prieto-Villalobos J., García-Rodríguez C., Sánchez N., Sáez J.C., Ortíz F.C, & Orellana J.A. (2024). Cx43 hemichannels and panx1 channels contribute to ethanol-induced astrocyte dysfunction and damage. Biological Research, 57, 15.

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Calcium Imaging of DRG Neurons

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Primary DRG neurons were dissociated at the L4 and L5 levels 14–16 days after inoculation and cultured as previously described.^{24 (link)} The cells were plated on glass coverslips coated with 0.1 mg/mL polyornithine and 1 mg/mL laminin (Boehringer Mannheim, Mannheim, Germany) and were incubated for 1 h at 37℃ with 5 μM Fura2-AM (Sigma) in HEPES-buffered Krebs solution (pH 7.4). The cells were washed and allowed to stand in a custom-designed perfusion chamber for 20 min at room temperature in the dark. Then, the chamber was mounted on the stage of an inverted microscope equipped with a CCD camera (Olympus, Japan). Cells were treated with α,β-meATP (20 μM) alone, or α,β-meATP (20 μM) followed by treatment with VPC32183 (20 μM), LPA (4 μM, Sigma), or BoTXC3 (5 µg/mL) and then α,β-meATP (20 μM). Fura-2 fluorescence signals were sequentially measured with excitation at 340 and 380 nm and emission at 510 nm to obtain a 340/380 ratio as a surrogate for intracellular calcium concentrations. Data were collected every 5 s. Images were acquired and analyzed using Metafluor imaging software (Universal Imaging Corp., Downingtown, PA, USA).

Wu J.X., Yuan X.M., Wang Q., Wei W, & Xu M.Y. (2016). Rho/ROCK acts downstream of lysophosphatidic acid receptor 1 in modulating P2X3 receptor-mediated bone cancer pain in rats. Molecular Pain, 12, 1744806916644929.

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Intracellular Sodium Measurement using SBFI

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Intracellular sodium concentration was measured using the sodium sensitive dye SBFI (Invitrogen) according to manufacturer’s instructions. In brief, 10nM stock solution of the dye in DMSO was further diluted in PBS (10ml) before loading onto the cells for 45 min. The signal was detected using a high-resolution digital camera (Photometrics Evolve, Roper Scientific). Excitation was provided by a Sutter DG-4 175W xenon arc lamp (Sutter Instruments). Regions of interest containing cells were selected using Metafluor imaging software (Universal Imaging). A FURA2 fluorescence lens kit was utilized to collect the fluorescent signal (Chroma Technology Corp, Bellows falls, VT). The ratio of fluorescent intensities at excitation 340nM/380nM/emission 510nM were recorded and the ratio calculated.

Kvirkvelia N., McMenamin M., Warren M., Jadeja R., Kodeboyina S.K., Sharma A., Zhi W., O’Conor P.M., Raju R., Lucas R, & Madaio M.P. (2018). Kidney targeted inhibition of protein kinase C-α ameliorates nephrotoxic nephritis with restoration of mitochondrial dysfunction. Kidney international, 94(2), 280-291.

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