Plan apochromat objective

Manufactured by Nikon

Sourced in Japan

The Plan-Apochromat objective is a high-performance lens designed for precision microscopy applications. It provides excellent optical performance, delivering uniform image quality across the entire field of view. The objective is engineered to minimize chromatic and spherical aberrations, ensuring accurate color reproduction and sharp, detailed images.

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Lab products found in correlation

Ti e inverted microscope, by Nikon (2 mentions) Ds qi1mc u3, by Nikon (2 mentions) Nis elements 4.3 acquisition software, by Nikon (2 mentions) Eclipse 80i, by Nikon (2 mentions) Ixonem 897e back illuminated emccd camera, by Oxford Instruments (1 mentions) Te2000 u inverted microscope, by Nikon (1 mentions) Elements software, by Nikon (1 mentions) μ slide, by Ibidi (1 mentions) Coolsnap es2 hq, by Nikon (1 mentions) Eclipse ti, by Nikon (1 mentions) Eclipse ti inverted microscope, by Nikon (1 mentions) Fluorescence mounting medium, by Agilent Technologies (1 mentions) Duo92101, by Merck Group (1 mentions) Ds qi2, by Nikon (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) Nis elements ar, by Nikon (1 mentions) Imagexpress micro confocal high content microscope, by Molecular Devices (1 mentions) 700 confocal microscope, by Zeiss (1 mentions) Zen imaging software, by Zeiss (1 mentions)

17 protocols using plan apochromat objective

Microscopic Imaging of Cellular Structures

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Images were captured using a Nikon TE-2000U inverted microscope (Nikon, Melville, NY) coupled to an iXonEM+ 897E back illuminated EMCCD camera (Andor, Belfast, U.K.) outfitted with a Bioptechs (Butler, PA) stage heater and 20x, Nikon Plan Apochromat objective. Bright field images were captured using Elements software (Nikon). For fluorescence microscopy, a xenon lamp illuminated cells through a 33-mm ND4 filter and 20x Nikon Plan Apochromat objective using a Nikon B2-A long pass emission filter set cube.

Johnson C.M., O’Brien X.M., Byrd A.S., Parisi V.E., Loosely A.J., Li W., Witt H., Faridi M.H., LeFort C.T., Gupta V., Kim M, & Reichner J.S. (2016). Integrin cross-talk regulates the human neutrophil response to fungal beta-glucan in the context of extracellular matrix: A prominent role for VLA3 in the anti-fungal response,. Journal of immunology (Baltimore, Md. : 1950), 198(1), 318-334.

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Live-cell Imaging of DiO-labeled Vesicles

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For live-cell imaging, cells were plated in 2-well slides (μ-slides; Ibidi, Madison, WI, USA) and imaged using a widefield system (Deltavision Core DV; GE Healthcare Life Sciences, Pittsburgh, PA, USA) at 37°C in a humidified environment with 5% CO₂. This system consists of an inverted microscope (IX71; Olympus) with a ×60 Plan-Apochromat objective (1.42 NA) and a camera (Coolsnap ES2 HQ; Nikon Corp., Tokyo, Japan). Images were captured every 2 minutes for a total of 4 hours on 3 z-planes and fluorescence was detected in the GFP (488 nm) and Cy5 (647 nm) channels. A differential interference contrast (DIC) image was captured at the central z-plane at each time point. Following acquisition, z-planes were stacked and the images were animated into a time-lapse movie at 12 frames per second. For some movies, the 488-nm channel was overlaid onto the DIC image. In addition, image analysis software (Imaris; Bitplane, Concord, MA, USA) was used to analyze the movies frame by frame and a dot was manually placed on DiO-labeled vesicles in each frame. The software then added a “dragon tail” to denote from where the vesicles had come.

Keller K.E., Bradley J.M., Sun Y.Y., Yang Y.F, & Acott T.S. (2017). Tunneling Nanotubes are Novel Cellular Structures That Communicate Signals Between Trabecular Meshwork Cells. Investigative Ophthalmology & Visual Science, 58(12), 5298-5307.

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Fluorescent Imaging of Cell Nuclei

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Fluorescence images were acquired using a Nikon Ti-E inverted microscope with a 20× (0.75 NA) Plan Apochromat objective with an optovar for an additional 1.5× magnification. DNA was detected using mCherry tagged histone H2B for cell lines and Hoechst staining for primary human neutrophils. All cell types were stained with Sytox Blue to determine viability. Viability scoring, nuclear segmentation and calculation of nuclear areas were carried out in MATLAB using custom software. Briefly, nuclei were segmented using an intensity threshold. A minimum area threshold of 100 pixels (~44uM) was implemented to filter out debris. Dead cells (nuclei positive for Sytox stain) were also excluded.

Rincón E., Rocha-Gregg B.L, & Collins S.R. (2018). A map of gene expression in neutrophil-like cell lines. BMC Genomics, 19, 573.

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Synchronizing HeLa Cells in G2 Phase

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HeLa cells transfected with GFP-H2B and other appropriate plasmids were synchronized at the G₁/S boundary by the double thymidine block method as described previously50 (link). At 8 h after release from the thymidine block, the cells were treated with 9 μM of RO3306 for 2 h to arrest cells at G₂ phase. The synchronized HeLa cells at G₂ phase regained the cell cycle progression in a microscope stage incubator at 37 °C in a humidified atmosphere of 5% CO₂ throughout the experiment. Fluorescence images were acquired every 5 min using a Nikon eclipse Ti with a 20 × 1.4 NA Plan-Apochromat objective. Images were captured with an iXonEM +897 Electron Multiplying charge-coupled device camera and analysed using NIS elements Ar microscope imaging software.

Lee H.S., Park Y.Y., Cho M.Y., Chae S., Yoo Y.S., Kwon M.H., Lee C.W, & Cho H. (2015). The chromatin remodeller RSF1 is essential for PLK1 deposition and function at mitotic kinetochores. Nature Communications, 6, 7904.

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Multiplexed Imaging with Quantum Dots

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Imaging was performed on an epi-fluorescence microscope (Nikon Ti Eclipse inverted microscope), using a planApo-chromat objective (60×, 1.45 N.A., Nikon). The samples were excited using DPSS lasers at 405 and 488 nm through a filter cube consisting of a quad band excitation filter (405, 488, 532, and 640 nm), a quad band dichroic filter, and a 505 long-pass emission filter (all from Chroma Technology). We used an additional emission filter for each QD such that the filter was a 20 nm band-pass centered at the nominal QD emission peak, except for the 705 QDs that used a 680 nm long-pass filter. Images were acquired using an EMCCD camera (iXon DV897, Andor Technologies) with an EM gain of 250 and an exposure time of 50 ms. For the multiplexing experiment in Figure 4B, we used an excitation laser at 405 nm and the emission filters 565/20, 610/40, and 685/70. We do observe ∼10% cross talk of QD565 and QD655 into the 610/40 nm filter, which is expected with this filter set, based on the spectral properties of these QDs.

Saurabh S., Beck L.E., Maji S., Baty C.J., Wang Y., Yan Q., Watkins S.C, & Bruchez M.P. (2014). Multiplexed Modular Genetic Targeting of Quantum Dots. ACS Nano, 8(11), 11138-11146.

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Fluorescence Microscopy Imaging Protocol

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Lipofectamine 2000 (Invitrogen) was used to transfect cells cultured on gelatin-coated coverslips with various constructs. After transfection, cells were incubated for 24 h, following which, they were fixed using 3.7% formaldehyde, and then permeabilized with phosphate-buffered saline (PBS) containing 0.1% Triton X-100 (PBST). Cells were treated with the blocking solution (1% BSA, 22.52 mg/mL glycine, and 0.1% gelatin in PBST), and were stained first with respective primary antibodies and subsequently with fluorophore-conjugated secondary antibodies diluted in blocking solution. Coverslips were mounted on microscope slides with fluorescence mounting medium (Dako) with or without Hoechst (Invitrogen). Images were acquired using a fluorescence microscope (Ti-E, Nikon) equipped with a 100× (1.4 N.A) Plan-Apochromat objective lens and a charge-coupled camera device (DS-Qi2, Nikon). The images were processed using NIS-Elements AR (Nikon) and/or ImageJ (NIH) software. A PLA assay was performed using a Duolink in situ red starter kit mouse/rabbit (Sigma-Aldrich, DUO92101), according to manufacturer’s instructions.

Yoon M.J., Choi B., Kim E.J., Ohk J., Yang C., Choi Y.G., Lee J., Kang C., Song H.K., Kim Y.K., Woo J.S., Cho Y., Choi E.J., Jung H, & Kim C. (2021). UXT chaperone prevents proteotoxicity by acting as an autophagy adaptor for p62-dependent aggrephagy. Nature Communications, 12, 1955.

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Immunofluorescence Microscopy Protocol

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Immunofluorescence was performed after fixing cells grown on glass coverslips, or directly in 96-well dish plates for high-throughput microscopy, for 20 min with 3% PFA in PBS. All steps of the immunofluorescence procedure were performed at room temperature. When indicated, cells were permeabilized for 5 min with 0.01% saponin in PBS before PFA fixation. After fixation, cells were incubated for 45 min in 1% fish skin gelatin and 0.1% saponin in PBS, followed by 30-min incubation with the primary antibody in 1% fish gelatin in PBS. After washing the primary antibody with PBS, the cells were incubated for 30 min with the secondary antibody (Cy2-, Cy3-, or Cy5-conjugated fluorescent antibodies) in 1% fish gelatin in PBS, followed by PBS washes. The cells were mounted in Mowiol 40-88 medium containing 10 µg/ml Hoechst and imaged with a Zeiss 700 confocal microscope (Carl Zeiss) using a 63× 1.4-NA oil differential interference contrast (DIC) Plan-Apochromat objective (Nikon) and Zen imaging software (Carl Zeiss). For high-throughput microscopy, 96-well plates were imaged with an ImageXpress Micro XLS Widefield High-content microscope (Molecular Devices) or an ImageXpress Micro Confocal High-content microscope (Molecular Devices; used in the wide-field mode) using a 40× 0.95-NA objective (Nikon).

Larios J., Mercier V., Roux A, & Gruenberg J. (2020). ALIX- and ESCRT-III–dependent sorting of tetraspanins to exosomes. The Journal of Cell Biology, 219(3), e201904113.

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Live-cell Imaging of CDC42 Dynamics

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dHL-60 cells expressing a previously characterized CDC42-FRET sensor (19 (link), 23 (link)) were resuspended in warmed modified L-15 media containing 2% FBS at a density of 500,000 cells/ml. To avoid washout, cells were plated on 96-well optical-glass bottomed imaging plates (Cellvis, catalog # P96–1.5H-N) coated with poly-D-lysine (“PDL”, Sigma; catalog # P6407). In a sterile tissue-culture hood, 50 μl of 200 mg/ml PDL was added to each well and incubated at room temperature for 30 minutes. After incubation, wells were washed twice with 50 μl sterile DPBS (Life Technologies; catalog # 14190250) and dried thoroughly at 65°C for at least 30 minutes before 100 μl of cells were added (results in ~40–50k cells/well). Before imaging, plated cells were incubated at 37°C for 30 minutes to allow adherence. Time-lapse microscopy was performed using a Nikon Ti-E inverted microscope with a 20x (0.75 NA) Plan Apochromat objective. Images were taken at 5 second intervals. A flash of red light immediately preceding the 4th frame was used to signal the timing for the addition of 100 μl of chemoattractant or media by pipet. To achieve rapid mixing, chemoattractants were added to media at a 1:1 ratio. Images were captured using 2×2 binning. For FRET experiments, Dual Zyla-4.2-USB3 sCMOS cameras were used to capture CFP and YFP channels simultaneously.

, & Weir E. (2000). Anthrax: of bison and bioterrorism. CMAJ: Canadian Medical Association Journal, 163(5), 608.

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Visualizing Cellular Vacuoles with CMAC

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Growing cells were laid on a thin layer of 1% agarose and viewed at room temperature with a fluorescence microscope (Eclipse E600; Nikon) equipped with a 100 differential interference contrast, numerical aperture (NA) 1.40 Plan-Apochromat objective (Nikon) and appropriate fluorescence light filter sets. Images were captured with a digital camera (DXM1200; Nikon) and ACT-1 acquisition software (Nikon) and processed with Photoshop CS (Adobe Systems). In each figure, we typically show only a few cells, representative of the whole population. Labeling of the vacuolar membrane with CMAC fluorescent dye was performed by adding 1 µl of the dye to 5 ml of culture at least 30 min prior to visualization.

Saliba E., Evangelinos M., Gournas C., Corrillon F., Georis I, & André B. (2018). The yeast H+-ATPase Pma1 promotes Rag/Gtr-dependent TORC1 activation in response to H+-coupled nutrient uptake. eLife, 7, e31981.

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Photoconversion of Kaede-labeled RPE

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Wild-type embryos were injected with Kaede mRNA. Embryos at 15 hpf with homogeneous green fluorescence were selected, mounted, and visualized under the Nikon AR1+ Confocal Microscope using a 20×/0.75 Plan-Apochromat objective. A region of interest (ROI) was drawn in the outer layer, corresponding to the putative position of the RPE progenitors, at a specific z-position and irradiated with the 405 nm laser at 21% of power for 10 loops to switch Kaede emission from green to red fluorescence. Due to confocality, photoconversion occasionally extended further than the selected plane, so that the tissues present above or below (i.e. ectoderm) also underwent photoconversion. After photoconversion embryos were let develop up to approximately 30 hpf stage, fixed and analysed by confocal microscopy for red fluorescence distribution.

Moreno-Mármol T., Ledesma-Terrón M., Tabanera N., Martin-Bermejo M.J., Cardozo M.J., Cavodeassi F, & Bovolenta P. (2021). Stretching of the retinal pigment epithelium contributes to zebrafish optic cup morphogenesis. eLife, 10, e63396.

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