Cfi plan apo vc

Manufactured by Nikon

Sourced in Japan

The CFI Plan Apo VC is a high-performance microscope objective lens from Nikon. It is designed to provide excellent optical performance, with features like aberration correction and high numerical aperture. The lens is intended for use in various scientific and research applications requiring high-quality imaging.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (3 mentions) Cfi plan apo 10 0, by Nikon (2 mentions) Plan apo 10 1, by Nikon (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Eclipse 90i, by Nikon (2 mentions) Ixon 897, by Oxford Instruments (1 mentions) Te2000 u, by Nikon (1 mentions) Nis element, by Nikon (1 mentions) Ixon ultra emccd camera, by Oxford Instruments (1 mentions) Cfi plan apochromat, by Nikon (1 mentions) Mlc 400 b laser box, by Agilent Technologies (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) Eclipse ti e inverted microscope, by Nikon (1 mentions) Ti e b microscope, by Nikon (1 mentions) Emccd camera, by Nikon (1 mentions) Metamorph software, by Molecular Devices (1 mentions) C9100 50 camera, by Hamamatsu Photonics (1 mentions) Ultraview vox spinning disk microscope, by Hamamatsu Photonics (1 mentions) Cytoseal 60, by Thermo Fisher Scientific (1 mentions) Temed, by Merck Group (1 mentions)

Close spelling variants of this product

CFI Plan Apo VC 20× CFI Plan Apo VC 100 CFI Plan Apo VC H 60x/1.4 CFI Plan Apo VC 60X WI Nikon CFI Plan Apo VC ×100/1.4 numerical aperture oil objective 100× CFI Plan Apo VC objective CFI Plan Apo VC 20X CFI plan Apo VC 60×, CFI Plan Apo VC 100X objective CFI Plan Apo VC 100X

15 protocols using cfi plan apo vc

Single-Molecule FRET Imaging Setup

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All FRET experiments were carried out on a custom-built prism-type TIRF setup built using a TE2000-U (Nikon) base. The sample was excited by an Ar/Kr ion laser (488 nm, Coherent), a Nd/YAG laser (532 nm, CrystaLaser) and a HeNe laser (633 nm, Laser 2000). Laser intensity as well as excitation duration (15–300 ms) was controlled by an acousto-optical tunable filter (AOTF.nC-TN, Pegasus Optik), which was synchronized to an EM-CCD camera (iXon+ 897, Andor) used for dual-color detection through a custom-built, real-time control unit. Fluorescence was collected by a 60× water immersion objective (CFI Plan Apo VC, Nikon, N.A. 1.2), spectrally separated by a dichroic mirror (630 DCXR, AHF) and filtered by the bandpass filters HQ550/88 and HQ715/150 (AHF). Fluorescence was detected on the EM-CCD camera split into two channels according to wavelength and an image series was recorded (Supplementary Figure S2A).

Heiss G., Ploetz E., Voith von Voithenberg L., Viswanathan R., Glaser S., Schluesche P., Madhira S., Meisterernst M., Auble D.T, & Lamb D.C. (2019). Conformational changes and catalytic inefficiency associated with Mot1-mediated TBP–DNA dissociation. Nucleic Acids Research, 47(6), 2793-2806.

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Immunocytochemistry of Transfected DRG

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Immunocytochemistry was performed on DRG that were trans-fected as described above. Briefly, cells were fixed using 4% paraformaldehyde, 4% sucrose for 20 minutes at RT. Permeabilization was achieved by 30 minutes incubation in PBS, 0.1% Triton X-100, following which nonspecific binding sites were saturated by PBS containing 3% BSA for 30 minutes. Cell staining was performed with primary antibodies in PBS with 3% BSA overnight at 4˚C. Cells were then washed 3 times in PBS and incubated with PBS containing 3% BSA and secondary antibodies (Alexa 488 goat anti-mouse and Alexa 594 goat anti-rabbit from Life Technologies) for 1 hour at RT. Immunofluorescent micrographs were acquired on a Nikon C1si scanning confocal microscope using CFI Plan APO VC 360 oil immersion objective with 1.4 numerical aperture. Camera gain and other relevant settings were kept constant. Colocalization analysis preformed with NiS Elements-Nikon (version 4.20, Nikon Instruments). Membrane-to-cytosol ratio was determined by defining regions of interest on the cytosol and the membrane of each cell using ImageJ. Total fluorescence was normalized to the area analyzed and before calculating the ratios.

Moutal A., Yang X., Li W., Gilbraith K.B., Luo S., Cai S., François-Moutal L., Chew L.A., Yeon S.K., Bellampalli S.S., Qu C., Xie J.Y., Ibrahim M.M., Khanna M., Park K.D., Porreca F, & Khanna R. (2017). CRISPR/Cas9 editing of Nf1 gene identifies CRMP2 as a therapeutic target in neurofibromatosis type 1-related pain that is reversed by (S)-Lacosamide. Pain, 158(12), 2301-2319.

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Confocal Microscopy for Axon Diameter

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Fluorescence confocal microscopy and image stack capturing were carried out as described in previous publications from our laboratory [27 (link),29 (link),30 (link),32 (link)]. In the present study, we used an Andor Revolution WD spinning disk confocal system (Oxford Instruments, Abingdon-on-Thames, UK) based on a Nikon TiE inverted microscope (60× CFI Plan Apo VC water immersion objective and numerical aperture at 1.40). We captured Z-stack images (8-bit TIFF files) at ~0.10 µm steps for the corpus callosum (Bregma between −1.82 and −2.30 mm) and cortical layer VI (Bregma between +1.98 and +1.78 mm). Image stacks were analyzed with NIH ImageJ. Axon diameters were measured on the Z-projection after Dynamic Reslice. The diameter values from the thickest and thinnest portions in the same axonal segment were compared and plotted.

Ma D., Deng B., Sun C., McComb D.W, & Gu C. (2022). The Mechanical Microenvironment Regulates Axon Diameters Visualized by Cryo-Electron Tomography. Cells, 11(16), 2533.

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Laser Scanning Confocal Microscopy of Cells

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Microscopy imaging was performed on a laser scanning confocal microscope (C1 si, Nikon) using a ×10 objective (CFI Plan Apochromat, Nikon) and a ×60 oil immersion objective (CFI Plan Apo VC, Nikon). The following lasers were used for excitation: a 405 nm continuous wave laser (Melles Griot 56ICS/S2695) for DAPI and Hoechst; a 488 nm continuous wave laser (Coherent Sapphire) for FD10, ATTO 488 and Alexa Fluor® 488; a 561 nm continuous wave laser (Melles Griot 85-YCA-010) for Alexa Fluor® 568 and a 640 nm continuous wave laser (Melles Griot 56ICS/S2695) for Alexa Fluor® 647. After photoporation, cells were incubated with 1 µg/mL Hoechst (Life Technology, Belgium) in CCM for 15 min at 37 °C. After the cells were washed twice with DPBS, time-lapse recordings were performed on a spinning disk confocal microscope (Nikon eclipse Ti-e inverted microscope, Nikon) equipped with an MLC 400 B laser box (Agilent technologies), a Yokogawa CSU-22 Spinning Disk scanner (Andor) and an iXon ultra EMCCD camera (Andor Technology, Belfast, UK). HeLa cells were imaged in a stage-top cell incubator (37°C with 5% CO₂ supplied, Tokai Hit) for 1 h with a time interval of 3 min using a ×60 oil immersion objective lens (CFI Plan Apo VC 60 × oil, Nikon, Japan).

Liu J., Xiong R., Brans T., Lippens S., Parthoens E., Zanacchi F.C., Magrassi R., Singh S.K., Kurungot S., Szunerits S., Bové H., Ameloot M., Fraire J.C., Teirlinck E., Samal S.K., Rycke R.D., Houthaeve G., De Smedt S.C., Boukherroub R, & Braeckmans K. (2018). Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy. Light, Science & Applications, 7, 47.

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Quantifying Cell Division Dynamics

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FIJI ImageJ was used to quantify the number of positive cells and the fluorescence intensity of Histone Nb labeled cells. 13 to 18 cells were quantified per data point of normalized mean fluorescence intensity (nMFI). The following exponential equation was used to obtain the cell division time τ:

nMFI = 2^{\land} (- (t / τ))

where t is imaging time in this work. The number of positive cells was quantified from 64 images taken with a 60×/1.4 Plan Apo VC Oil immersion objective (CFI Plan Apo VC, Nikon).

Liu J., Hebbrecht T., Brans T., Parthoens E., Lippens S., Li C., De Keersmaecker H., De Vos W.H., De Smedt S.C., Boukherroub R., Gettemans J., Xiong R, & Braeckmans K. (2020). Long-term live-cell microscopy with labeled nanobodies delivered by laser-induced photoporation. Nano research, 13(2), 485-495.

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TIRF Microscopy of Lifeact or Vinculin Cells

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For TIRF microscopy, Lifeact or vinculin-transfected cells were imaged using a NIKON TI-E/B microscope, equipped with an Evolve EMCCD camera. The illumination system used was from from Roper Scientific (iLas²), and the objective was a Nikon CFI Plan APO VC oil immersion objective (60X, N.A. 1.4). The microscope was controlled with MetaMorph software (Molecular Devices). Temperature, CO₂, and humidity control was performed using a chamlide TC system (TC-A, Quorum technologies). The laser used for excitation of EGFP was a 491 nm cobolt Calypso laser, and the one for excitation of mCHerry was a 561 nm MPBC Green Visible Fiber laser. ET525/50M (Chroma) and ET605/52M (Chroma) emission filters were used for for EGFP and mCherry fluorescence collection, respectively.

Golovkine G., Faudry E., Bouillot S., Elsen S., Attrée I, & Huber P. (2016). Pseudomonas aeruginosa Transmigrates at Epithelial Cell-Cell Junctions, Exploiting Sites of Cell Division and Senescent Cell Extrusion. PLoS Pathogens, 12(1), e1005377.

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Dendritic and Axonal Arborization Imaging

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Neurons (25–30 DIV) were imaged at room temperature in modified Tyrode solution (pH 7.4) on a Perkin-Elmer UltraView VoX Spinning Disk microscope with an autofocus system, a motorized stage, 488 nm/50 mW and 561 nm/50 mW diode lasers and a Hamamatsu C9100-50 camera. This system included a PhotoKinesis feature for fluorescence recovery after photobleaching that was used in the case of photoswitching experiments. All experiments were performed using a 60X/1.4 numerical aperture (NA) oil immersion objective lens (CFI Plan Apo VC, Nikon), and images were acquired at 4 frames/s. One dendritic and one axonal arborization were randomly selected to perform each experiment.

An S.J., Stagi M., Gould T.J., Wu Y., Mlodzianoski M., Rivera-Molina F., Toomre D., Strittmatter S.M., De Camilli P., Bewersdorf J, & Zenisek D. (2022). Multimodal imaging of synaptic vesicles with a single probe. Cell Reports Methods, 2(4), 100199.

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Photobleaching Assay for Fluorescent Proteins

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We constructed a photobleaching chamber using a glass microscope slide (catalog no. 7101, Henso) as the base and then mounting a coverslip (catalog no. 48366-227, VWR) on top of two additional spacer coverslips attached to the base with tape. We then mixed a 1 μM dilution of each FP with 20% (w/v) acrylamide/bis-acrylamide, 30% (w/v) solution (catalog no. A3699, Sigma-Aldrich), 3 μl of 10% ammonium persulfate (catalog no. A3678, Sigma-Aldrich), and 0.5 μl of N,N,N′,N′-tetramethylethylenediamine (TEMED) (catalog no. T9281, Sigma-Aldrich) in a 1.5-ml tube. Right after adding TEMED, we transferred ~80 μl of the solution to the space between the top coverslip and glass slide and waited ~5 min for the gel to polymerize. We then sealed the edges with Cytoseal 60 (catalog no. 8310-4, Thermo Fisher Scientific). After 12 hours, the slides were continuously photobleached for 7 min with 5-s-interval image captures using widefield 510/25-nm light with 62.5 mW/mm² (using a 60× 1.4-NA oil objective; CFI Plan Apo VC, Nikon Instruments). For each FP, nine fields of view were tested. For each field of view, the photobleaching half-life was calculated by determining the time at which the fluorescence reached half of the initial fluorescence. The mean photobleaching half-life values were calculated and plotted.

Lee J., Liu Z., Suzuki P.H., Ahrens J.F., Lai S., Lu X., Guan S, & St-Pierre F. (2020). Versatile phenotype-activated cell sorting. Science Advances, 6(43), eabb7438.

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Immunofluorescence Staining of Lung Tissues

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Lung tissues were collected from mice that had been sacrificed under a CO₂ atmosphere. They were filled with PBS (−):optimal cutting temperature (OCT) compound (1:1), and the resulting lung tissues were embedded in OCT compound. The resulting tissues were sliced at a thickness of 5 μm. After placing the slices on MAS-coated glass slides (Matsunami, Tokyo, Japan), they were then incubated with the first antibody solution using a Cy3-labeled anti-αSMA antibody (Sigma-Aldrich, C6198), anti-mouse CD31 Armenian Hamster immunoglobulin G (IgG) (MA31505, Thermo Fisher Scientific), anti-mouse VEcad rat IgG (138001, BioLegend), and anti-bromodeoxyuridine (BrdU) mouse IgG (364101, BioLegend). The slices were then washed twice with PBS (−) and immersed in a second antibody solution containing DyLight488-labeled anti-Armenian hamster IgG goat IgG (405503, BioLegend) for CD31 and AlexaFluor647-labeled anti-Rat IgG goat IgG (A21247, Thermo Fisher Scientific) for 30 min. The slices were covered with a coverslip (Matsunami, 1s) after washing with PBS (−). The tumor slices were observed with a Nikon A1R system (Nikon, Tokyo, Japan) using CFI Plan Apo Lambda 20× or CFI Plan Apo VC 60× water immersion objective lens.

Sakurai Y., Hada T., Kato A., Hagino Y., Mizumura W, & Harashima H. (2018). Effective Therapy Using a Liposomal siRNA that Targets the Tumor Vasculature in a Model Murine Breast Cancer with Lung Metastasis. Molecular Therapy Oncolytics, 11, 102-108.

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Confocal Microscopy Imaging of FITC-Dextran

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Confocal microscopy imaging was performed on a laser scanning confocal microscope (C2, Nikon, Tokoyo, Japan) using a 10× magnification (CFI Plan Apo VC, Nikon, Japan). The 488 nm continuous wave laser (Coherent Sapphire, Menlo Par, CA, USA) was used for excitation of FITC-dextran.

Liu J., Li C., Brans T., Harizaj A., Van de Steene S., De Beer T., De Smedt S., Szunerits S., Boukherroub R., Xiong R, & Braeckmans K. (2020). Surface Functionalization with Polyethylene Glycol and Polyethyleneimine Improves the Performance of Graphene-Based Materials for Safe and Efficient Intracellular Delivery by Laser-Induced Photoporation. International Journal of Molecular Sciences, 21(4), 1540.

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