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Planapo 60 1.42 na objective

Manufactured by Olympus
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The PlanApo 60× 1.42 NA objective is a high-numerical aperture, plan-apochromatic objective lens designed for advanced microscopy applications. It provides a magnification of 60× and a numerical aperture of 1.42, enabling high-resolution imaging and excellent light-gathering capabilities.

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Lab products found in correlation

Dv elite cmos camera, by Olympus (6 mentions) Deltavision microscope, by Cytiva (3 mentions) γh2ax, by Merck Group (3 mentions) C4742 95, by Hamamatsu Photonics (2 mentions) Axioplan 2 microscope, by Hamamatsu Photonics (2 mentions) Rad51, by Merck Group (2 mentions) Sc 137021, by Santa Cruz Biotechnology (2 mentions) 1.40 na objective, by Olympus (2 mentions) Fluoview fv1000, by Olympus (1 mentions) Igor pro, by Wavemetrics (1 mentions) P1530, by Merck Group (1 mentions) A11070, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

PlanApo 60×, (14 citations) PlanApo N 60×/1.42 NA oil objective (6 citations) 60× 1.42 NA objective (5 citations) PlanApo N 60×/NA 1.42 objective (3 citations) ×60 NA 1.42 objective (2 citations) PlanApo NA 1.35 (2 citations) PlanApo 60×/1.42 NA oil immersion objective (2 citations)

9 protocols using planapo 60 1.42 na objective

1

Telomeric FISH and CO-FISH Analysis

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Telomeric FISH and CO-FISH were conducted as previously described (van Steensel et al., 1998 (link); Celli et al., 2006 (link)) using Alexa Fluor 647-OO-(TTAGGG)3, Cy3-OO-(TTAGGG)three or FITC-OO-(CCCTAA)three and a centromere probe (PNA Bio). Images were captured using a DeltaVision microscope (Applied Precision) equipped with a cooled charge-coupled device camera (DV Elite CMOS Camera) and a PlanApo 60 × 1.42 NA objective (Olympus America), and controlled by and SoftWoRx software.
Schmutz I., Mensenkamp A.R., Takai K.K., Haadsma M., Spruijt L., de Voer R.M., Choo S.S., Lorbeer F.K., van Grinsven E.J., Hockemeyer D., Jongmans M.C, & de Lange T. (2020). TINF2 is a haploinsufficient tumor suppressor that limits telomere length. eLife, 9, e61235.
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2

Immunofluorescence Imaging of DNA Repair Factors

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Previously published procedures were followed for IF and IF-FISH12 (link). IF for myc-tagged RPA32 or Ctc1 (mouse monoclonal, 9B11 or rabbit monoclonal, 71D10, Cell Signaling Technology), HA-tagged Stn1 (3724, Cell Signaling Technology), endogenous Polα (sc-137021, Santa Cruz), and 53BP1 (612522, BD Biosciences) was carried out using the cytoskeleton extraction protocol14 (link). Intensity measurements of RPA32-myc IF were performed in FIJI as follows: nuclei were identified using thresholding, segmented, and identified as regions of interest. The average image background was then subtracted from the image, and the total raw pixel intensity within each area of interest in the channel of interest was calculated. Rad51 (70-001, Bioacademia), and γH2AX (05636, Millipore) were detected in cells fixed in 3% PFA, and foci showing co-localization of Rad51 with γH2AX were quantified. IF imaging was performed on a Zeiss Axioplan II microscope equipped with a Hamamatsu C4742-95 camera using Volocity software or on a DeltaVision (Applied Precision) equipped with a cooled charge-coupled device camera (DV Elite CMOS Camera), a PlanApo 60× 1.42 NA objective or 100× 1.40 NA objective (Olympus America, Inc.), and SoftWoRx software.
Mirman Z., Lottersberger F., Takai H., Kibe T., Gong Y., Takai K., Bianchi A., Zimmermann M., Durocher D, & de Lange T. (2018). 53BP1/Rif1/Shieldin counteract DSB resection through CST/Polα-dependent fill-in. Nature, 560(7716), 112-116.
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3

Immunofluorescence Imaging of DNA Repair Factors

Cited 1 time
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Previously published procedures were followed for IF and IF-FISH12 (link). IF for myc-tagged RPA32 or Ctc1 (mouse monoclonal, 9B11 or rabbit monoclonal, 71D10, Cell Signaling Technology), HA-tagged Stn1 (3724, Cell Signaling Technology), endogenous Polα (sc-137021, Santa Cruz), and 53BP1 (612522, BD Biosciences) was carried out using the cytoskeleton extraction protocol14 (link). Intensity measurements of RPA32-myc IF were performed in FIJI as follows: nuclei were identified using thresholding, segmented, and identified as regions of interest. The average image background was then subtracted from the image, and the total raw pixel intensity within each area of interest in the channel of interest was calculated. Rad51 (70-001, Bioacademia), and γH2AX (05636, Millipore) were detected in cells fixed in 3% PFA, and foci showing co-localization of Rad51 with γH2AX were quantified. IF imaging was performed on a Zeiss Axioplan II microscope equipped with a Hamamatsu C4742-95 camera using Volocity software or on a DeltaVision (Applied Precision) equipped with a cooled charge-coupled device camera (DV Elite CMOS Camera), a PlanApo 60× 1.42 NA objective or 100× 1.40 NA objective (Olympus America, Inc.), and SoftWoRx software.
Mirman Z., Lottersberger F., Takai H., Kibe T., Gong Y., Takai K., Bianchi A., Zimmermann M., Durocher D, & de Lange T. (2018). 53BP1/Rif1/Shieldin counteract DSB resection through CST/Polα-dependent fill-in. Nature, 560(7716), 112-116.
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4

Quantifying Synaptic Protein Levels in C. elegans

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To control for protein expression levels in the extrachromosomal arrays, animals were first immobilized using 2,3-butanedione monoxime (30 mg/ml, Alfa Aesar, Ward Hill, MA), mounted on 2% agarose pads, in M9 buffer (22.0 mM KH2PO4, 42.3 mM Na2HPO4, 85.6 mM NaCl, and 1.0 mM MgSO4), and imaged on an inverted Olympus microscope (IX81), using a laser scanning confocal imaging system (Olympus Fluoview FV1000 with dual confocal scan heads) and an Olympus PlanApo 60× 1.42 NA objective. Rescuing complexin variants were C-terminally tagged with GFP separated by a 12 residue linker (GGSGGSGGSAAA), and synaptic protein levels were estimated by measuring background-subtracted fluorescence between dorsal cord synaptic peaks. Data were analyzed with custom software in IGOR Pro (WaveMetrics, Lake Oswego, OR; Burbea et al., 2002 (link); Dittman and Kaplan, 2006). A fluorescent slide was imaged daily to monitor the laser stability and the dorsal cord fluorescence was normalized to the slide value.
Radoff D.T., Dong Y., Snead D., Bai J., Eliezer D, & Dittman J.S. (2014). The accessory helix of complexin functions by stabilizing central helix secondary structure. eLife, 3, e04553.
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5

Immunofluorescence with Telomeric FISH

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For immunofluorescence in combination with telomeric FISH (IF-FISH), cells grown on coverslips to sub-confluence and were fixed in MeOH for 10 min at −20°C. IF-FISH was carried out as previously described (Takai et al., 2003 (link)). The following affinity purified antibodies were used for IF: rabbit TRF2 (#647), rabbit TRF1 (#371), rabbit TIN2 2 (#864), rabbit 53BP1 (Abcam ab175933). Telomeric DNA was detected with FITC-OO-(CCCTAA)3 PNA probe. Images were captured on a DeltaVision microscope (Applied Precision) equipped with a cooled charge-coupled device camera (DV Elite CMOS Camera), a PlanApo 60 × 1.42 NA objective (Olympus America), and SoftWoRx software.
Schmutz I., Mensenkamp A.R., Takai K.K., Haadsma M., Spruijt L., de Voer R.M., Choo S.S., Lorbeer F.K., van Grinsven E.J., Hockemeyer D., Jongmans M.C, & de Lange T. (2020). TINF2 is a haploinsufficient tumor suppressor that limits telomere length. eLife, 9, e61235.
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6

Quantifying DNA Damage Response Markers

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Previously published procedures were followed for IF (17 (link)) using γH2AX (Millipore), 53BP1 (BD), RIF1, BRCA1, BARD1, RAD51 antibodies. Secondaries used were anti-mouse highly cross-absorbed alexa fluor plus 488 and anti-rabbit highly cross-absorbed alexa fluor plus 647 (Thermo Fisher). Imaging was performed on a DeltaVision (Applied Precision) equipped with a cooled charge-coupled device camera (DV Elite CMOS Camera), a PlanApo 60× 1.42 NA objective (Olympus America, Inc.), and SoftWoRx software. Deconvoluted images were max projected before scoring foci manually or with automated foci counting software (FIJI). For 53BP1 cavity scoring, samples were blinded from the investigators, and foci were determined to be either cavity-containing, globular, or ambiguous. After excluding ambiguous foci, the percent of foci containing a cavity was derived from the total foci scored.
Mirman Z., Sharma K., Carroll T.S, & de Lange T. (2022). Expression of BRCA1, BRCA2, RAD51, and other DSB repair factors is regulated by CRL4WDR70. DNA repair, 113, 103320.
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7

High-Resolution Epifluorescence Imaging

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High quality/high resolution imaging was performed on a GE Healthcare DVLive epifluorescence image restoration microscope using an Olympus PlanApo 60×/1.42 NA objective and a 1.9k × 1.9k sCMOS camera. Z stacks (0.25 μm) covering the whole nucleus (∼10 μm) were acquired before applying a conservative restorative algorithm for quantitative image deconvolution. Max intensity projections were generated and used for image analysis.
Stossi F., Dandekar R.D., Mancini M.G., Gu G., Fuqua S.A., Nardone A., De Angelis C., Fu X., Schiff R., Bedford M.T., Xu W., Johansson H.E., Stephan C.C, & Mancini M.A. (2020). Estrogen-induced transcription at individual alleles is independent of receptor level and active conformation but can be modulated by coactivators activity. Nucleic Acids Research, 48(4), 1800-1810.
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8

Quantifying MAVS Spatial Distribution

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LM2 and SUM159 cells were treated with DMSO, 20nM H3B-8800, or .5ug/mL p (I:C) (Sigma Aldrich, P1530). High resolution imaging was performed on a GE Healthcare DVLive epifluorescence image restoration microscope using an Olympus PlanApo 60×/1.42 NA objective and a 1.9k × 1.9x sCMOS camera. Z stacks (0.25μm) were acquired before applying a conservative restorative algorithm for quantitative image deconvolution. Max intensity projections were generated and used for image analysis. To quantify the spatial distribution of the MAVS signal, we measured a dispersion index using a custom-made MATLAB script. Briefly, z-stacks were transformed into a 2D image by maximal projection. Single cells were then manually segmented using an interactive polygonal ROI. The dispersion index of the MAVS signal for each cell was then determined as follows: first, the MAVS signal was segmented using an Otsu thresholding method, and the weighted centroid location of the resulting mask was determined using the underlying pixel intensities. Then, the distance between each pixel of the MAVS mask to the weighted centroid was calculated and the dispersion index was defined as the mean of these distances. Aggregation was calculated as the inverse of the dispersion index.
Bowling E.A., Wang J.H., Gong F., Wu W., Neill N.J., Kim I.S., Tyagi S., Orellana M., Kurley S.J., Dominguez-Vidaña R., Chung H.C., Hsu T.Y., Dubrulle J., Saltzman A.B., Li H., Meena J.K., Canlas G.M., Chamakuri S., Singh S., Simon L.M., Olson C.M., Dobrolecki L.E., Lewis M.T., Zhang B., Golding I., Rosen J.M., Young D.W., Malovannaya A., Stossi F., Miles G., Ellis M.J., Yu L., Buonamici S., Lin C.Y., Karlin K.L., Zhang X.H, & Westbrook T.F. (2021). Spliceosome-Targeted Therapies Trigger an Antiviral Immune Response in Triple-Negative Breast Cancer. Cell, 184(2), 384-403.e21.
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9

Immunofluorescence Analysis of 53BP1 Foci

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Cells were grown on glass coverslips and fixed in 3% paraformaldehyde and 2% sucrose. Coverslips were permeabilized in 0.5% Triton-X-100/PBS, and blocked in goat block (0.1% BSA, 3% goat serum, 0.1% Triton-X-100, 2 mM EDTA) in PBS. Primary and secondary antibodies (Rabbit anti-53BP1 Abcam #ab-175933 1:1000, F(ab’)2-goat anti-rabbit IgG (H + L) Cross-adsorbed Alexa Fluor 488 ThermoFisher A-11070, 1:500) were diluted in goat block. Slides were counter-stained with DAPI and mounted using prolong gold antifade medium. Images were acquired on a DeltaVision microscope (Applied Precision) equipped with a cooled charge-coupled device camera (DV Elite CMOS Camera), with a PlanApo ×60  1.42 NA objective (Olympus America), and SoftWoRx software. Images were analyzed for foci numbers using a custom-made algorithm written for FIJI, courtesy of Leonid Timashev51 (link).
Dewhurst S.M., Yao X., Rosiene J., Tian H., Behr J., Bosco N., Takai K.K., de Lange T, & Imieliński M. (2021). Structural variant evolution after telomere crisis. Nature Communications, 12, 2093.
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