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1.49 na tirf objective

Manufactured by Olympus
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The 60 × 1.49 NA TIRF objective is a high-numerical aperture objective designed for total internal reflection fluorescence (TIRF) microscopy. It has a magnification of 60× and a numerical aperture of 1.49, which allows for efficient light collection and high-resolution imaging of fluorescently labeled samples near the coverslip surface.

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Lab products found in correlation

Ixon887, by Oxford Instruments (1 mentions) Ix70 microscope, by Olympus (1 mentions) Cage incubator, by Okolab (1 mentions) Emccd camera, by Olympus (1 mentions) Prime95b cmos camera, by Teledyne (1 mentions) Eclipse ti2 microscope, by Nikon (1 mentions) Stage top incubator, by Okolab (1 mentions) Ixon 897 emccd camera, by Oxford Instruments (1 mentions) μ slide 8 well glass bottom, by Ibidi (1 mentions) Till photonics imic tirf microscope, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

TIRF NA 1.49 DIC objective 60 × 1.49 NA objective

4 protocols using 1.49 na tirf objective

1

Multi-Angle TIRFM Imaging of Cells

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A custom-built multi-angle TIRFM system was used to image the cells12 (link),13 (link). This TIRFM setup was based on an Olympus IX-70 microscope, equipped with 405-, 488- and 568-nm laser lines, a 60 × 1.49 NA TIRF objective (Olympus) and an EMCCD camera (iXon887; Andor Technology). The angles of the illumination laser beam can be rapidly adjusted by two-dimensional galvometers under the control of custom-written software, which allows for both Epi- and TIRF imaging of cells. For imaging of live cells, TIRF images were acquired under the control of Andor iQ software at 0.2–0.5 Hz with exposure times in the rage of 200–500 ms.
Zhou X., Wang J., Chen J., Qi Y., Di N.a.n., Jin L., Qian X., Wang X., Chen Q., Liu X, & Xu Y. (2018). Optogenetic control of epithelial-mesenchymal transition in cancer cells. Scientific Reports, 8, 14098.
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2

Live-cell TIRF Microscopy of PI(3,4,5)P3 Dynamics

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Live-cell imaging was done as previously described (16 (link)) using a custom TIRF microscope equipped with 488- and 568-nm solid state lasers (Melles Griot), an EMCCD camera, and a 60 × 1.49 NA TIRF objective (Olympus). All experiments were done at 37°C (using a custom incubator chamber) in phenol red-free DMEM with 25 mM HEPES, pH 7.4, with or without 10% FBS. Image frames were acquired in time-lapse recordings at 2 or 0.2 Hz with 150-ms exposures. Pixel size was 160 nm. For PI(3,4,5)P3 imaging with PHAKT–stagRFP, cells were imaged at 37°C and 5% CO2 in a cage incubator (OkoLab) housing a Nikon Eclipse Ti2 microscope (Nikon) equipped with a motorized Ti-LA-HTIRF module with 15-mW LU-N4 488- and 561-nm lasers, using a CFI Plan Apochromat Lambda 100×/1.45 Oil TIRF objective and a Prime95B cMOS camera (110-nm pixel size; Teledyne Photometrics). Images were acquired using a 100-ms exposure time at 1 Hz.
An S.J., Anneken A., Xi Z., Choi C., Schlessinger J, & Toomre D. (2022). Regulation of EGF-stimulated activation of the PI-3K/AKT pathway by exocyst-mediated exocytosis. Proceedings of the National Academy of Sciences of the United States of America, 119(48), e2208947119.
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3

TIRF Microscopy of NPY-mRFP and LifeAct-GFP

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RBL-CXCR1 cells transiently cotransfected with NPY-mRFP and LifeAct-GFP were seeded at 2 × 105 cells per chamber in 8-well chamber borosilicate coverglass systems (Ibidi μ-slide 8-well glass bottom, catalog no. 80827). The next day, cells were activated under the microscope, and images were acquired withaZeiss510 (Zeiss, Oberkochen, Germany) or Leica SP5 microscope equipped with a HyD detector by using a ×63 oil/1.4 NA objective equipped with a top-stage incubator (Okolab, Ottaviano, Italy) set to 37°C and 5% CO2.
For total internal reflection fluorescence (TIRF) microscopy, images were acquired with a TILL Photonics iMIC TIRF microscope (FEI, Munich, Germany) using an Olympus 100× NA 1.49 TIRF objective. Images were acquired with an Andor iXon 897 EMCCD camera (Andor, Belfast, United Kingdom), and the imaging protocol was controlled by using TILL Photonics Live Acquisition software. TIRF angle was set to provide minimal penetration of the evanescent wave while still producing measurable signal from the LifeAct and NPY structures. We used the 360° TIRF feature, which azimuthally spins the laser beam on the circumference of the objective back focal plane, creating homogenous TIRF illumination across the field. Data were analyzed by using ImageJ software (National Institutes of Health, Bethesda, Md).
Klein O., Krier-Burris R.A., Lazki-Hagenbach P., Gorzalczany Y., Mei Y., Ji P., Bochner B.S, & Sagi-Eisenberg R. (2019). Mammalian diaphanous-related formin 1 (mDia1) coordinates mast cell migration and secretion through its actin-nucleating activity. The Journal of allergy and clinical immunology, 144(4), 1074-1090.
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4

Live-cell Imaging of RABV and NGF

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DRG explants were imaged with TILL Photonics iMIC TIRF microscope (FEI, Munich, Germany), using Olympus 100× NA 1.49 TIRF objective. Images were acquired with an Andor iXon 897 EMCCD camera (Belfast, UK) and imaging protocol was controlled by TILL Photonics LiveAcquisition software. Throughout the experiment, explants were imaged inside an environmental chamber maintaining 37°C and 5% CO2. TIRF angle was set to give minimal penetration of the evanescent wave still giving measurable signal from the RABV-EGFP/NGF-QD particles. We used the 360° TIRF feature which azimuthally spins the laser beam on the circumference of the objective back focal plane, creating homogenous TIRF illumination across the field. Broad neuronal tips were selected for imaging after RABV/NGF addition, selected fields were imaged for 5–10 minutes each, for up to 2 hours. Exposure times were 50 msec, and camera gain was set to 300. Final frame rate was 1 and 1.12 seconds per frame in RABV/NGF alone and RABV+p75NTR experiments, respectively. A frame is comprised of RABV/NGF/p75 fluorescent channels over a bright-field image to localize fluorescent particles in context. For RABV-p75 imaging, explant cultures were incubated with fluorescent anti-p75NTR (ANT-007-AO, Alomone Labs) for 10 minutes and washed 3 times in poor neurobasal medium prior to imaging.
Gluska S., Zahavi E.E., Chein M., Gradus T., Bauer A., Finke S, & Perlson E. (2014). Rabies Virus Hijacks and Accelerates the p75NTR Retrograde Axonal Transport Machinery. PLoS Pathogens, 10(8), e1004348.
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