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Cfi apo tirf 100

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The CFI Apo TIRF 100X is an objective lens designed for total internal reflection fluorescence (TIRF) microscopy. It provides a high numerical aperture of 1.49 and a magnification of 100X, enabling high-resolution imaging of samples near the coverslip surface.

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Lab products found in correlation

Perfect focus system, by Nikon (6 mentions) Zt488 10, by Chroma Technology (4 mentions) Zet640 20, by Chroma Technology (4 mentions) Zt488rdc zt561rdc zt640rdc, by Chroma Technology (4 mentions) Et525 50m, by Chroma Technology (4 mentions) Et700 75m, by Chroma Technology (4 mentions) Et600 50m, by Chroma Technology (4 mentions) Zet561 10, by Chroma Technology (4 mentions) Tirf illuminator, by Nikon (2 mentions) Eclipse ti, by Nikon (2 mentions) Zt561rdc, by Chroma Technology (2 mentions) Et575lp, by Chroma Technology (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Alexa fluor 647 conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 555 conjugated donkey anti goat igg, by Thermo Fisher Scientific (1 mentions) A1r confocal microscope, by Nikon (1 mentions) Alexa fluor 488 conjugated chicken anti rabbit, by Thermo Fisher Scientific (1 mentions) Eclipse ti e, by Nikon (1 mentions) Electrically heated chamber, by Okolab (1 mentions)

Spelling variants of this product

CFI Apo TIRF (10 citations) Nikon CFI Apo TIRF 100× 1.49 N.A. oil objective (7 citations) CFI Apo TIRF 100×H/1.49 (6 citations) CFI Apo 100× (5 citations) CFI Apo TIRF ×100 oil objective (4 citations) CFI Apo TIRF 100× 1.49 NA objective (4 citations) CFI Apo TIRF 100× Oil (2 citations) CFI Apo TIRF 100×/1.49-NA oil-immersion objective (2 citations)

8 protocols using cfi apo tirf 100

1

High-Resolution Fluorescence Imaging of Biological Samples

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Fluorescence imaging was carried out on an inverted Nikon Eclipse Ti microscope (Nikon Instruments) with the Perfect Focus System, applying an objective-type TIRF configuration using a Nikon TIRF illuminator with an oil-immersion objective (CFI Apo TIRF × 100, NA 1.49, Oil) yielding a pixel size of 160 nm. Two lasers were used for excitation—488 nm (200 mW nominal, Coherent Sapphire) and 647 nm (300 mW nominal, MBP Communications). The laser beam was passed through clean-up filters (ZT488/10 and ZET640/20, Chroma Technology) and coupled into the microscope objective using a multi-band beam splitter (ZT488rdc/ZT561rdc/ZT640rdc, Chroma Technology). Fluorescence light was spectrally filtered with emission filters (ET525/50 m and ET700/75 m, Chroma Technology) and imaged on an EMCCD camera (iXon X3 DU-897, Andor Technologies). Images were acquired with a CCD readout bandwidth of 3 MHz at 14 bit, 5.1 pre-amp gain and no electron-multiplying gain using the centre 256 × 256 px of the CCD chip. Imaging was performed using highly inclined and laminated optical sheet illumination39 (link) with an excitation intensity of ∼50 mW using the 647 nm laser line. A total of 15,000 frames at a frame rate of 10 Hz were collected, resulting in ∼25 min imaging time.
Beliveau B.J., Boettiger A.N., Avendaño M.S., Jungmann R., McCole R.B., Joyce E.F., Kim-Kiselak C., Bantignies F., Fonseka C.Y., Erceg J., Hannan M.A., Hoang H.G., Colognori D., Lee J.T., Shih W.M., Yin P., Zhuang X, & Wu C.T. (2015). Single-molecule super-resolution imaging of chromosomes and in situ haplotype visualization using Oligopaint FISH probes. Nature Communications, 6, 7147.
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2

Multicolor TIRF Microscopy Imaging

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Fluorescence imaging was carried out on an inverted Nikon Eclipse Ti microscope (Nikon Instruments) with the Perfect Focus System, applying an objective-type TIRF configuration using a Nikon TIRF illuminator with an oil-immersion objective (CFI Apo TIRF 100×, NA 1.49, Oil). For 2D imaging an additional 1.5 magnification was used to obtain a final magnification of ≈150-fold, corresponding to a pixel size of 107 nm. Three lasers were used for excitation: 488 nm (200 mW nominal, Coherent Sapphire), 561 nm (200 mW nominal, Coherent Sapphire) and 647 nm (300 mW nominal, MBP Communications). The laser beam was passed through cleanup filters (ZT488/10, ZET561/10, and ZET640/20, Chroma Technology) and coupled into the microscope objective using a multi-band beam splitter (ZT488rdc/ZT561rdc/ZT640rdc, Chroma Technology). Fluorescence light was spectrally filtered with emission filters (ET525/50m, ET600/50m, and ET700/75m, Chroma Technology) and imaged on an EMCCD camera (iXon X3 DU-897, Andor Technologies).
Jungmann R., Avendano M.S., Woehrstein J.B., Dai M., Shih W.M, & Yin P. (2014). Multiplexed 3D Cellular Super-Resolution Imaging with DNA-PAINT and Exchange-PAINT. Nature methods, 11(3), 313-318.
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3

Single-molecule TIRF Microscopy Protocol

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Fluorescence imaging was carried out on an inverted Nikon Eclipse Ti microscope (Nikon Instruments) with the Perfect Focus System, applying an objective-type TIRF configuration using a Nikon TIRF illuminator with an oil-immersion objective (CFI Apo TIRF 100×, NA 1.49, Oil) yielding a pixel size of 160 nm. Two lasers were used for excitation: 488 nm (200 mW nominal, Coherent Sapphire) and 647 nm (300 mW nominal, MBP Communications). The laser beam was passed through cleanup filters (ZT488/10 and ZET640/20, Chroma Technology) and coupled into the microscope objective using a multi-band beam splitter (ZT488rdc/ZT561rdc/ZT640rdc, Chroma Technology). Fluorescence light was spectrally filtered with emission filters (ET525/50m and ET700/75m, Chroma Technology) and imaged on an EMCCD camera (iXon X3 DU-897, Andor Technologies). Images were acquired with a CCD readout bandwidth of 3 MHz at 14 bit, 5.1 pre-amp gain and no electron-multiplying gain using the center 256×256 px of the CCD chip. Imaging was performed using HILO illumination39 (link) with an excitation intensity of ~50 mW using the 647 nm laser line. A total of 15,000 frames at a frame rate of 10 Hz were collected, resulting in ~25 min imaging time.
Beliveau B.J., Boettiger A.N., Avendaño M.S., Jungmann R., McCole R.B., Joyce E.F., Kim-Kiselak C., Bantignies F., Fonseka C.Y., Erceg J., Hannan M.A., Hoang H.G., Colognori D., Lee J.T., Shih W.M., Yin P., Zhuang X, & Wu C.T. (2015). Single-molecule super-resolution imaging of chromosomes and in situ haplotype visualization using Oligopaint FISH probes. Nature Communications, 6, 7147.
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4

Multicolor TIRF Microscopy Imaging

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Fluorescence imaging was carried out on an inverted Nikon Eclipse Ti microscope (Nikon Instruments) with the Perfect Focus System, applying an objective-type TIRF configuration using a Nikon TIRF illuminator with an oil-immersion objective (CFI Apo TIRF 100×, NA 1.49, Oil). For 2D imaging an additional 1.5 magnification was used to obtain a final magnification of ≈150-fold, corresponding to a pixel size of 107 nm. Three lasers were used for excitation: 488 nm (200 mW nominal, Coherent Sapphire), 561 nm (200 mW nominal, Coherent Sapphire) and 647 nm (300 mW nominal, MBP Communications). The laser beam was passed through cleanup filters (ZT488/10, ZET561/10, and ZET640/20, Chroma Technology) and coupled into the microscope objective using a multi-band beam splitter (ZT488rdc/ZT561rdc/ZT640rdc, Chroma Technology). Fluorescence light was spectrally filtered with emission filters (ET525/50m, ET600/50m, and ET700/75m, Chroma Technology) and imaged on an EMCCD camera (iXon X3 DU-897, Andor Technologies).
Jungmann R., Avendano M.S., Woehrstein J.B., Dai M., Shih W.M, & Yin P. (2014). Multiplexed 3D Cellular Super-Resolution Imaging with DNA-PAINT and Exchange-PAINT. Nature methods, 11(3), 313-318.
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5

Imaging Differentiated Human Myotubes

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Differentiated human myotubes grown on a glass coverslip coated with laminin were fixed with 4% paraformaldehyde (made in PBS), permeabilized with 1% Triton in PBS for 20 min, and processed as previously described (Treves et al., 2011 (link)). The following antibodies were used: mouse anti-RyR1 (MA3-925; Thermo Fisher Scientific), goat anti-Cav1.1 (sc-8160; Santa Cruz Biotechnology, Inc.), rabbit anti-Cav1.2 (sc-25686; Santa Cruz Biotechnology, Inc.), mouse anti-dystrophin (ab-7164; Abcam), rabbit anti-utrophin (sc-15377; Santa Cruz Biotechnology, Inc.), Alexa Fluor 488–conjugated chicken anti–rabbit, Alexa Fluor 555–conjugated donkey anti–goat IgG (Thermo Fisher Scientific), and Alexa Fluor 647–conjugated goat anti–mouse IgG (Thermo Fisher Scientific). Cells were stained with 4′, 6-diamidino-2-phenylindole (DAPI; Thermo Fisher Scientific) to visualize nuclei and observed using an A1R confocal microscope (Nikon) with a CFI Apo TIRF 100× (1.49 NA) objective.
Sekulic-Jablanovic M., Ullrich N.D., Goldblum D., Palmowski-Wolfe A., Zorzato F, & Treves S. (2016). Functional characterization of orbicularis oculi and extraocular muscles. The Journal of General Physiology, 147(5), 395-406.
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6

Single-Molecule Live-Cell Imaging

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All single-molecule assays were carried out on an inverted microscope (Nikon Eclipse Ti-E) fitted with a CFI Apo TIRF 100 × oil-immersion objective (NA 1.49, Nikon). The temperature was maintained at 31.2°C by an electrically heated chamber (Okolab). dsDNA was visualized every 10 s for 30 min by exciting the S.O. with a 568-nm laser (Coherent, Sapphire 568–200 CW) at 80 mW/cm2. The red fluorescently labeled proteins were excited at 80 mW/cm2 (800 W/cm2 during a FRAP pulse) with a 647-nm laser (Coherent, Obis 647–100 CW). The AF488–Pol ε was visualized with a 488-nm laser (Coherent, Sapphire 488–200 CW) at 140 mW/cm2. The signals were spectrally separated using appropriate filter sets (Chroma) and fluorescence signals collected on an Evolve 512 Delta EMCCD (Photometics). Typically, nine fields of view (five for the FRAP experiments) were selected for imaging. Single-molecule experimental results were derived from at least three or four technical replicates for each experimental condition.
Lewis J.S., Spenkelink L.M., Schauer G.D., Yurieva O., Mueller S.H., Natarajan V., Kaur G., Maher C., Kay C., O’Donnell M.E, & van Oijen A.M. (2019). Tunability of DNA Polymerase Stability during Eukaryotic DNA Replication. Molecular cell, 77(1), 17-25.e5.
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7

DNA-PAINT Imaging with Gold Fiducials

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Before imaging, 90 nm gold nanoparticle fiducial markers were added and incubated for 5 min (diluted 1:5 in PBS + 5 mM MgCl2). After rinsing with PBS with 5 mM MgCl2, imaging buffer containing DNA-PAINT imaging strands was added. DNA-PAINT imaging was carried out on an inverted Nikon Eclipse Ti microscope (Nikon Instruments) with the Perfect Focus System, applying an objective-type TIRF configuration with an oil-immersion objective (CFI Apo TIRF 100×, NA 1.49, Oil). Samples were excited using a 561 nm laser (200 mW nominal, Coherent Sapphire). The laser beam was passed through a cleanup filter (ZET561/10, Chroma Technology) and coupled into the microscope objective using a beam splitter (ZT561rdc, Chroma Technology). Fluorescence light was spectrally filtered with two emission filters (ET600/50m and ET575lp, Chroma Technology) and imaged on an sCMOS camera (Zyla 4.2, Andor Technologies). Imaging was performed without additional magnification in the detection path and 2×2 camera binning resulting in a pixel size of 130 nm.
Strauss S., Nickels P.C., Strauss M.T., Sabinina V.J., Ellenberg J., Carter J.D., Gupta S., Janjic N, & Jungmann R. (2018). Modified aptamers enable quantitative sub-10-nm cellular DNA-PAINT imaging. Nature methods, 15(9), 685-688.
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8

DNA-PAINT Imaging with Gold Fiducials

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Before imaging, 90 nm gold nanoparticle fiducial markers were added and incubated for 5 min (diluted 1:5 in PBS + 5 mM MgCl2). After rinsing with PBS with 5 mM MgCl2, imaging buffer containing DNA-PAINT imaging strands was added. DNA-PAINT imaging was carried out on an inverted Nikon Eclipse Ti microscope (Nikon Instruments) with the Perfect Focus System, applying an objective-type TIRF configuration with an oil-immersion objective (CFI Apo TIRF 100×, NA 1.49, Oil). Samples were excited using a 561 nm laser (200 mW nominal, Coherent Sapphire). The laser beam was passed through a cleanup filter (ZET561/10, Chroma Technology) and coupled into the microscope objective using a beam splitter (ZT561rdc, Chroma Technology). Fluorescence light was spectrally filtered with two emission filters (ET600/50m and ET575lp, Chroma Technology) and imaged on an sCMOS camera (Zyla 4.2, Andor Technologies). Imaging was performed without additional magnification in the detection path and 2×2 camera binning resulting in a pixel size of 130 nm.
Strauss S., Nickels P.C., Strauss M.T., Sabinina V.J., Ellenberg J., Carter J.D., Gupta S., Janjic N, & Jungmann R. (2018). Modified aptamers enable quantitative sub-10-nm cellular DNA-PAINT imaging. Nature methods, 15(9), 685-688.
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