Plan apochromat 1.3 na oil immersion lens

Manufactured by Zeiss

The 40x Plan-apochromat 1.3 NA oil immersion lens is a high-performance optical lens designed for use in microscopy applications. It features a magnification of 40x and a numerical aperture of 1.3, which allows for the capture of high-resolution, detailed images. This lens is optimized for use with oil immersion techniques, which can improve image quality and resolution. The plan-apochromatic design of the lens helps to minimize chromatic and spherical aberrations, resulting in sharp, well-corrected images across the entire field of view.

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Lab products found in correlation

Lsm 700 microscope, by Zeiss (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Aqua poly mount, by Polysciences (2 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Duolink anti mouse minus and anti rabbit plus in situ pla probes, by Merck Group (1 mentions) Duolink in situ detection reagents red, by Merck Group (1 mentions)

Close spelling variants of this product

63× oil immersion lens 63/1.4 NA Plan Apochromat Plan-Apochromat

4 protocols using plan apochromat 1.3 na oil immersion lens

Visualizing UV-Induced DNA Damage and Protein Recruitment

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Cells were grown on 18 mm coverslips, fixed in 4% paraformaldehyde, and permeabilized in PBS containing 0.5% Triton X-100. For visualization of local UV-induced DNA damage (LUD), DNA was denatured for 5 min with 70 mM NaOH. Next, cells were incubated in blocking buffer (3% BSA and 2.25% glycine in PBS-T (0.1% Tween 20)) for 1 h at room temperature. Primary antibodies were incubated for 1–2 h at room temperature or overnight at 4 °C and secondary antibodies conjugated to Alexa fluorochromes 488 or 555 (Invitrogen) were incubated for 1 h at room temperature. The antibody incubation solution was 1% BSA in PBS-T. DNA was stained with DAPI (Sigma), and slides were mounted using Aqua-Poly/Mount (Polysciences, Inc.). Antibodies used are summarized in Supplementary Tables 1 and 2. Image acquisition was performed using an LSM700 microscope equipped with a 40x Plan-apochromat 1.3 NA oil immersion lens (Carl Zeiss Micro Imaging Inc.). To quantify protein recruitment to lesion sites, the fluorescence signal intensity at LUD was divided by the nuclear intensity, as measured using FIJI image analysis software (version 1.52p). Zero accumulation (nuclear background) was set to 0 and maximum accumulation (above nuclear background) in control or mock-treated conditions was set at 1.0.

Ribeiro-Silva C., Sabatella M., Helfricht A., Marteijn J.A., Theil A.F., Vermeulen W, & Lans H. (2020). Ubiquitin and TFIIH-stimulated DDB2 dissociation drives DNA damage handover in nucleotide excision repair. Nature Communications, 11, 4868.

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Immunofluorescence Staining of DNA Damage

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Cells were grown on 24-mm glass coverslips and fixed for 15 min in PBS supplemented with 2% formaldehyde and 0.1% Triton X-100. Subsequently, cells were permeabilized with PBS containing 0.1% Triton X-100 and blocked with 2% bovine serum albumin (BSA) in PBS for 30 minutes at RT. For staining DNA damage with lesion-specific antibodies, DNA was denatured with 70 mM NaOH in PBS for 10 min. Thereafter, cells were incubated with the appropriate antibodies diluted in PBS overnight at 4 °C. Cells were washed with PBS containing 0.1% Triton X-100 and incubated with AlexaFluor™-conjugated secondary antibodies (Invitrogen) and DAPI for 1 hour at RT. After repeated washing with PBS including 0.1% Triton X-100, cells were mounted using Aqua-Poly/Mount and digital images were acquired using an LSM700 microscope equipped with a 40x Plan-apochromat 1.3 NA oil immersion lens (Carl Zeiss). Fluorescence intensities were determined using Fiji and an ImageJ macro. Fold accumulation of the respective proteins were calculated by dividing the average fluorescent intensity at LUD through the average fluorescent intensity inside the rest of the nucleus. Primary and secondary antibodies used for immunofluorescence are listed in the Key Resources Table, respectively.

van Toorn M., Turkyilmaz Y., Han S., Zhou D., Kim H.S., Salas-Armenteros I., Kim M., Akita M., Wienholz F., Raams A., Ryu E.J., Kang S., Theil A.F., Bezstarosti K., Tresini M., Giglia-Mari G., Demmers J.A., Schärer O.D., Choi J.H., Vermeulen W, & Marteijn J.A. (2022). Active DNA damage eviction by HLTF stimulates nucleotide excision repair. Molecular cell, 82(7), 1343-1358.e8.

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Proximity Ligation Assay for Protein Interactions

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Cells were grown on 12 mm coverslips and fixed with 3.6% formaldehyde in PBS for 15 minutes. Cells were permeabilized with PBS containing 0.1% Triton X-100, blocked with 2% BSA in PBS for 30 minutes at RT, and incubated with the appropriate antibodies overnight at 4 °C. PLA was performed using the Duolink anti-Mouse MINUS and anti-Rabbit PLUS In Situ PLA probes (Sigma) and the Duolink In Situ Detection Reagents Red (Sigma), according to the to manufacturer’s instructions. Digital images were acquired using an LSM700 microscope equipped with a 40x Plan-apochromat 1.3 NA oil immersion lens (Carl Zeiss) and nuclear foci were quantified using an ImageJ macro.

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Immunostaining Protocol for DNA Damage

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Cells were grown on coverslips, fixed in 4% paraformaldehyde and permeabilized in PBS containing 0.5% Triton X-100. DNA was denatured for 5 min with 70 mM NaOH to allow CPD binding by the antibody. Next, cells were incubated for 1 h with blocking solution 3% BSA in PBS-T (0.1% Tween 20) and subsequently incubated with antibodies diluted in 1% BSA with PBS-T (0.1% Tween 20) for 1–2 h at room temperature or overnight at 4 °C. To visualize primary antibodies, cells were incubated for 1 h at room temperature with secondary antibodies conjugated to Alexa fluorochromes 488, 555, or 633 (Invitrogen). DNA was stained with DAPI (Sigma), and slides were mounted using Aqua-Poly/Mount (Polysciences, Inc.). Antibodies used are summarized in Supplementary Tables 1 and 2. Images were acquired using an LSM700 microscope equipped with a 40x Plan-apochromat 1.3 NA oil immersion lens (Carl Zeiss Micro Imaging Inc.). Using FIJI image analysis software, we determined protein accumulation at lesion sites by dividing the overall fluorescence signal intensity at LUDs by the protein overall nuclear intensity. In Fig. 1f and Fig. 6g zero accumulation (nuclear background) was set at 0 and maximum accumulation (above nuclear background) in control conditions at 1.0.

Ribeiro-Silva C., Aydin Ö.Z., Mesquita-Ribeiro R., Slyskova J., Helfricht A., Marteijn J.A., Hoeijmakers J.H., Lans H, & Vermeulen W. (2018). DNA damage sensitivity of SWI/SNF-deficient cells depends on TFIIH subunit p62/GTF2H1. Nature Communications, 9, 4067.

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