Plan apochromat

Manufactured by Hamamatsu Photonics

Sourced in United States, France

The Plan-Apochromat is an optical lens designed for use in microscopy. It is a type of objective lens that provides high-quality, distortion-free images with excellent color correction across the visible spectrum. The Plan-Apochromat is known for its flat field of view and uniform sharpness, making it well-suited for a variety of microscopy applications.

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Lab products found in correlation

Metamorph 7, by Molecular Devices (2 mentions) Cell observer sd microscope, by Yokogawa (1 mentions) C13440, by Hamamatsu Photonics (1 mentions) Alexa fluor secondary antibody, by Thermo Fisher Scientific (1 mentions) Polyvinyl alcohol (pva), by Merck Group (1 mentions) Dmr microscope, by Leica (1 mentions) Orca 100, by Hamamatsu Photonics (1 mentions) Flash 4, by Hamamatsu Photonics (1 mentions) Illustrator cs6, by Adobe (1 mentions) Orca flash4.0 v2, by Hamamatsu Photonics (1 mentions) Plan apochromat, by Zeiss (1 mentions) Anti fade dapi fluoromount g, by Southern Biotech (1 mentions) Axio observer a1, by Zeiss (1 mentions) V3 confocal spinning disk head 89 north, by Hamamatsu Photonics (1 mentions) Orcafusion geniii scmos camera, by Hamamatsu Photonics (1 mentions) Orca flash 4.0 lt plus digital cmos camera, by Hamamatsu Photonics (1 mentions) Te2000 e pfs, by Nikon (1 mentions) Pen strep, by Thermo Fisher Scientific (1 mentions) Incucyte live cell imaging system, by Sartorius (1 mentions) Phenol red, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Plan-Apochromat 20×/0.8 objective (4 citations) Plan-Apochromat objective (4 citations) Plan-Apochromat 63X phase (3 citations) Plan-Apochromat 100 × /1.4 oil DIC objective (3 citations) Plan-Apochromat 206/0.8 M27 air objective (3 citations) Plan Apochromat 100× 1.45 NA oil objective (2 citations) 63x 1.4NA Plan Apochromat (2 citations) CFI60 Plan Apochromat Lambda 40× Objectives (2 citations) Plan-Apochromat oil objective (2 citations) Plan Apochromat 100 × /1.4 oil phase objective (2 citations)

9 protocols using plan apochromat

Imaging Mouse Embryonic Stem Cells

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For the experiment in Fig 1D, mouse ES cells expressing DONSON‐GFP and mCherry‐PSF1 from the endogenous loci were grown on “μ‐Slide 4‐well” (Ibidi, 80426) with “no phenol red DMEM medium” (ThermoFisher Scientific, 21063029) supplemented as described above. For p97 inhibition, cells were treated with 5 μM CB‐5083 for 3 h before imaging.
Confocal images of live cells were acquired with a Zeiss Cell Observer SD microscope with a Yokogawa CSU‐X1 spinning disk, using a HAMAMATSU C13440 camera with a PECON incubator, a 60× 1.4‐NA Plan‐Apochromat oil immersion objective, and excitation and emission filter sets for GFP and mCherry. Images of live mouse ES cells were acquired using “ZEN blue” software (Zeiss) and processed with ImageJ software (National Institutes of Health) as previously described (Sonneville et al, 2017 (link)).

Evrin C., Alvarez V., Ainsworth J., Fujisawa R., Alabert C, & Labib K.P. (2023). DONSON is required for CMG helicase assembly in the mammalian cell cycle. EMBO Reports, 24(11), e57677.

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Immunofluorescence Microscopy Quantification

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Cells grown on glass coverslips were washed once with PBS and fixed using 4% PFA or ice-cold methanol. Cells were then permeabilized with 0.2% Triton X-100 in PBS. Primary antibodies were added to coverslips and incubated at 37°C for 1 h followed by multiple washes in PBS. Alexa Fluor secondary antibodies (Thermo Fisher Scientific) were added and incubated for 30 min, followed by PBS washes and mounting of coverslips in polyvinyl alcohol (Sigma-Aldrich). Samples were visualized using a DMR microscope (Leica Microsystems) fitted with 40-Å (NA 1.0, Plan Fluotar) and 63-Å (NA 1.32, Plan Apochromat) objective lenses, and images were captured with an Orca 100 charge-coupled device camera (model C4742-95; Hamamatsu Photonics) and MetaMorph 7.7 imaging software (Molecular Devices). Mean fluorescence pixel intensity was quantified using raw, unsaturated 12-bit images with line or area tracing as stated using ImageJ. For quantification of junctional/ID fluorescence intensity, cell borders as marked by Pg signal using ImageJ line tracing and fluorescence intensity of Pg and the costained protein were assessed from the same line position. Subsequently, signal intensity values of the protein of interest were normalized to Pg intensity.

Kam C.Y., Dubash A.D., Magistrati E., Polo S., Satchell K.J., Sheikh F., Lampe P.D, & Green K.J. (2018). Desmoplakin maintains gap junctions by inhibiting Ras/MAPK and lysosomal degradation of connexin-43. The Journal of Cell Biology, 217(9), 3219-3235.

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Visualizing SETD2 RNA Localization in ccRCC

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FG2 ccRCC cells were transiently transfected with a wt SETD2-GFP expression plasmid (Carvalho et al., 2014 (link)) and cultured on glass coverslip for 24 hr before hybridization with RNA FISH probes (Biosearch Technologies, CA, USA) following the manufacturer’s protocol. Briefly, cells were washed with PBS, fixed for 10 min at room temperature, washed twice with PBS, and permeabilized at 4°C in 70% (vol/vol) EtOH. Probes diluted in hybridization buffer were added to permeabilized cells before overnight incubation in a dark chamber at 37°C. After washing, DAPI was added to stain the nuclei. Epi-fluorescence microscopy was performed using a Zeiss Z1 microscope equipped with Z-piezo (Prior, MA, USA), a 63x 1.4 NA Plan-Apochromat objective and a sCMOS camera (Hamamatsu Flash 4.0). FISH probes were designed to target a segment of the RNA transcript encoded by the intergenic region downstream of the canonical termination sites of MRPL23 and SEL1L3. The sequences of the probes are shown in Supplementary file 1J.

Grosso A.R., Leite A.P., Carvalho S., Matos M.R., Martins F.B., Vítor A.C., Desterro J.M., Carmo-Fonseca M, & de Almeida S.F. (2015). Pervasive transcription read-through promotes aberrant expression of oncogenes and RNA chimeras in renal carcinoma. eLife, 4, e09214.

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Immunofluorescence Staining Protocol

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Cells were fixed with phosphate-buffered saline containing 4% paraformaldehyde (for 15 min), excepting that for the staining of ATR, cells were fixed with 100% methanol (at -20°C for 10 min). Cells were permeabilized with 0.1% Triton X-100 (for 60 min) or 0.5% Triton X-100 (for 20 min) after methanol- or paraformaldehyde-fixation, respectively. The fixed cells were stained with primary antibodies for 1 h and then incubated with Alexa Fluor 488- or 546-conjugated secondary antibodies or phalloidin for 1 h. Cells were mounted on the slides with mounting medium (Anti-Fade Dapi-Fluoromount-G, Southern Biotech). The images (2048x2048 pixels) were acquired using an upright fluorescence microscope (Axio imager, M2 Apotome2 system, Carl Zeiss), which was equipped with 63x and 100x oil-immersion objective lens (1.4 NA, Plan Apochromat) and a camera (ORCA-Flash4.0 V2; Hamamatsu). For Figure S8, images were acquired by a Carl Zeiss LSM880 confocal microscope imaging system with a Zeiss Plan-Apochromat 63x/NA 1.4 oil immersion objective. The representative images were cropped by Photoshop CS6 (Adobe) and assembled by Illustrator CS6 (Adobe).

Fan J.R., Chang S.N., Chu C.T, & Chen H.C. (2023). AKT2-mediated nuclear deformation leads to genome instability during epithelial-mesenchymal transition. iScience, 26(6), 106992.

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Neuronal Imaging and Ascaroside Stimulation

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Neuronal imaging was performed using a Zeiss AxioObserver.A1 inverted microscope with Zeiss Plan-Apochromat objective lens (10x/0.45 NA) and a Hamamatsu Orca Flash 4.0 sCMOS camera mounted with a 1.0x C-mount adapter. MicroManager software was used to acquire image stacks (10 frames s⁻¹ for 30 s per trial) and to control liquid stimulus delivery (5 – 15 s) via a microscope controller (Nobska Imaging) that actuated Parker solenoid valves via a Automate ValveLink 8.2 controller. Excitation illumination pulses (10 ms per frame) were delivered from a 50W blue LED (Mightex) or Lumencor SOLA through an EGFP filter set. For testing individual ascarosides in sequence, the inlet tube was transferred manually to each ascaroside tube and allowed to flow for 1 min between stimuli.

Fagan K.A., Luo J., Lagoy R.C., Schroeder F.C., Albrecht D.R, & Portman D.S. (2018). A single-neuron chemosensory switch determines the valence of a sexually dimorphic sensory behavior. Current biology : CB, 28(6), 902-914.e5.

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Optimized DAPI Staining Protocol

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DAPI staining was done by modifying standard protocols (Francis and Nayack, 2000), with the cold methanol fixation done for a shorter time (2.5 min) and the concentration of DAPI higher at 1 μg/ml in 0.01% Tween in PBS in the dark for 5 min, washed once with 0.1% Tween in PBS. Samples were briefly stored at 4°C in 75% glycerol and imaged directly in glycerol solution on a Leica DMI8 with an xLIGHT V3 confocal spinning disk head (89 North) with a ×63 Plan-Apochromat (1.4 NA) objective and an ORCAFusion GenIII sCMOS camera (Hamamatsu Photonics) controlled by microManager (Edelstein et al., 2010). DAPI was excited with a 405 nm laser. Worms were mounted on agar pads in 75% glycerol.

Misare K.R., Ampolini E.A., Gonzalez H.C., Sullivan K.A., Li X., Miller C., Sosseh B., Dunne J.B., Voelkel-Johnson C., Gordon K.L, & Hartman J.H. (2023). The consequences of tetraploidy on Caenorhabditis elegans physiology and sensitivity to chemotherapeutics. bioRxiv.

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Time-Lapse Imaging of Frz-Signaling Modulation

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Protocol followed is as reported previously by Guzzo et al. [11 (link)]. For this assay, cells were grown at 32°C in CYE medium. After overnight incubation, 1 ml of cells was centrifuged for 5 minutes at 7,000 rpm, and pellets were resuspended in TPM medium (10 mM Tris [pH 7.6], 8 mM MgSO₄, 1 mM KH₂PO₄) to OD₆₀₀ of 2 units. A total of 2 μl of cells were spotted on coverslip below a TPM 1.5% agar pad with addition of 0.075% IAA in melted agar before pouring the pad, allowing modulation of Frz-signaling intensity with IAA. IAA solutions were made in TPM buffer containing 1mM CaCl₂. After 10 minutes of incubation at room temperature, time-lapse experiments were performed using an automated and inverted epifluorescence microscope TE2000-E- PFS (Nikon, France), with a 40x/0.75 DLL “Plan-Apochromat” objective and an ORCA-Flash4.0 LT PLUS Digital CMOS camera (#C11440-42U30 Hamamatsu Photonics). The microscope is equipped with a PFS that automatically maintains focus. Images were recorded with NIS-Eléments AR 4.60 software (Nikon).

Baranwal J., Lhospice S., Kanade M., Chakraborty S., Gade P.R., Harne S., Herrou J., Mignot T, & Gayathri P. (2019). Allosteric regulation of a prokaryotic small Ras-like GTPase contributes to cell polarity oscillations in bacterial motility. PLoS Biology, 17(9), e3000459.

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Live-cell imaging of mitotic duration

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To quantify mitotic duration and cell division status (Fig1B and C), cells were seeded in 12-well plates (Nunc). An hour prior to imaging, the medium was changed to CO₂-independent medium without phenol red (Invitrogen) supplemented with 10% FCS, 0.2 mM l-glutamine, PenStrep, and 1 mM Na-pyruvate (all Invitrogen). Phase contrast images of cells were acquired every 5 min at 37°C using a Zeiss Axio Observer Z1 microscope controlled by SimplePCI software (Hamamatsu) equipped with an Orca 03GO1 camera (Hamamatsu) and a Plan-Apochromat 10×/0.45 objective.
To quantify mitotic duration for Supplementary Fig S8A, cells were reverse transfected with the indicated siRNAs in 24-well Imagelock plates (Essen BioScience). 12 h after seeding, the plate was imaged using an Incucyte live cell imaging system (Essen BioScience). Images were processed using ImageJ software and analysed using the mitotic duration plugin. For high resolution imaging (Fig1D), HeLa Kyoto cells stably expressing H2B-mCherry and α-tubulin-GFP were seeded in Labtek chambers (Nunc, Thermo Scientific) and imaged at intervals of 5 min using a Zeiss Axio Observer Z1 microscope controlled by SimplePCI software (Hamamatsu) with a 40× /1.3 DIC H oil objective.

Sundaramoorthy S., Vázquez-Novelle M.D., Lekomtsev S., Howell M, & Petronczki M. (2014). Functional genomics identifies a requirement of pre-mRNA splicing factors for sister chromatid cohesion. The EMBO Journal, 33(22), 2623-2642.

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Fluorescence Microscopy Imaging of Chromosomal Structures

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Images were acquired with a microscope stand (Eclipse TE2000-U; Nikon) with a 100× Plan Apochromat, 1.4 NA 100× digital interference contrast oil immersion lens with a camera (ORCA-ER; Hamamatsu Photonics) at room temperature (25°C). MetaMorph 7.1 (Molecular Devices) was used to acquire unbinned images of in a seven-step z series with a 300-nm step size of Smc4-GFP, kinetochore proteins Nuf2-GFP and Cse4-2×GFP, 12.5-kb LacO, Tfc1-GFP, Cbf5-GFP, and Lrs4-GFP. Images were taken with 600-ms exposure times for RFP and GFP in water on 0.135-mm coverslips. Images of 1.7-kb LacO arrays in tDNAΔ and WT were acquired in 10 200-nm steps. Population images of the CEN15 LacO/LacI-GFP and CEN11 TetO/TetR-CFP strain were binned 2× and acquired in 10 200-nm steps, whereas time-lapse images were acquired in single planes at 15- and 30-s time intervals. Live cells were imaged in synthetic growth medium.

Snider C.E., Stephens A.D., Kirkland J.G., Hamdani O., Kamakaka R.T, & Bloom K. (2014). Dyskerin, tRNA genes, and condensin tether pericentric chromatin to the spindle axis in mitosis. The Journal of Cell Biology, 207(2), 189-199.

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