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Hcx pl apo 63 1.40 0 60 oil λ bl objective

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The HCX PL APO 63×/1.40–0.60 OIL λ BL objective is a high-performance microscope objective designed for Leica microscopes. It provides a magnification of 63x and a numerical aperture range of 1.40 to 0.60, making it suitable for use with oil immersion techniques. The objective is part of the Leica PL APO series and features a lambda (λ) corrected optical design and a brightline (BL) coating for optimal performance.

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Lab products found in correlation

Hcx plan fluotar 10 0.30 na objective, by Nikon (2 mentions) Te2000 microscope, by Nikon (2 mentions) Glass bottom dishes, by MatTek (1 mentions)

5 protocols using hcx pl apo 63 1.40 0 60 oil λ bl objective

1

3D CDM Migration Assay Protocol

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Three-dimensional CDMs were generated as previously described13 (link). The thickness of the generated CDMs was measured using a Leica SP5 scanning confocal with a HCX PL APO 63×/1.40–0.60 OIL λ BL objective. The orientation of the fibronectin stained fibers was quantified using the OrientationJ plugin for ImageJ52 (link). The modal angle, as defined as the angle that had the highest percentage of fibers oriented, was set to 0 to allow for direct comparison between images.
For migration analysis, parental MDA-MB-231 human breast cancer cells (ATCC) were seeded into the generated CDMs for 4 hours in the presence of serum. After 4 hours, time lapse imaging was performed on a Nikon TE2000 microscope equipped with an environmental chamber and imaged using a HCX Plan Fluotar 10×/0.30 NA objective with images taken every 10 minutes for 16 hours. The percentage of cells exhibiting plasticity was assessed by scoring the number of cells undergoing at least one amoeboid to mesenchymal phenotypic switch. The Manual tracking plugin in ImageJ was used to track the cell centroid over the length of the movie. The Chemotaxis and Migration plugin in ImageJ was used to calculate directionality and migration velocity.
Goreczny G.J., Ouderkirk-Pecone J.L., Olson E.C., Krendel M, & Turner C.E. (2016). Hic-5 Remodeling of the Stromal Matrix Promotes Breast Tumor Progression. Oncogene, 36(19), 2693-2703.
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2

Adhesion Dynamics Analysis with Vinculin and Zyxin

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For adhesion dynamics analysis, vinculin-YFP or zyxin-GFP expressing cells were transfected with a control siRNA or siRNAs targeting cdGAP. Cells were re-plated onto soft or hard PDMS-coated MatTek glass bottom dishes (MatTek Corp, Ashland, MA) overnight before being imaged on a Leica SP5 confocal microscope using an HCX PL APO 63×/1.40–0.60 OIL λ BL objective (Leica, Bannockburn, IL) equipped with an environment chamber maintained at 37°C with regulated CO2. Time-lapse movies were compiled and background subtracted before being analyzed in ImageJ. Only individual adhesions that could be followed from the point at which they were initially visible all the way through until complete disassembly were quantified for lifetime analysis.
Wormer D.B., Davis K.A., Henderson J.H, & Turner C.E. (2014). The Focal Adhesion-Localized CdGAP Regulates Matrix Rigidity Sensing and Durotaxis. PLoS ONE, 9(3), e91815.
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3

Quantifying Fibronectin Matrix Clearance

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CAFs were plated onto 10μg/mL fibronectin coated glass coverslips and allowed to spread/clear the fibronectin matrix for 4 hours. Images of the cells were acquired using a Leica SP5 scanning confocal with a HCX PL APO 63×/1.40–0.60 OIL λ BL objective. Area of fibronectin fibers was quantified relative to the cell area using ImageJ.
Goreczny G.J., Ouderkirk-Pecone J.L., Olson E.C., Krendel M, & Turner C.E. (2016). Hic-5 Remodeling of the Stromal Matrix Promotes Breast Tumor Progression. Oncogene, 36(19), 2693-2703.
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4

3D CDM Migration Assay Protocol

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Three-dimensional CDMs were generated as previously described13 (link). The thickness of the generated CDMs was measured using a Leica SP5 scanning confocal with a HCX PL APO 63×/1.40–0.60 OIL λ BL objective. The orientation of the fibronectin stained fibers was quantified using the OrientationJ plugin for ImageJ52 (link). The modal angle, as defined as the angle that had the highest percentage of fibers oriented, was set to 0 to allow for direct comparison between images.
For migration analysis, parental MDA-MB-231 human breast cancer cells (ATCC) were seeded into the generated CDMs for 4 hours in the presence of serum. After 4 hours, time lapse imaging was performed on a Nikon TE2000 microscope equipped with an environmental chamber and imaged using a HCX Plan Fluotar 10×/0.30 NA objective with images taken every 10 minutes for 16 hours. The percentage of cells exhibiting plasticity was assessed by scoring the number of cells undergoing at least one amoeboid to mesenchymal phenotypic switch. The Manual tracking plugin in ImageJ was used to track the cell centroid over the length of the movie. The Chemotaxis and Migration plugin in ImageJ was used to calculate directionality and migration velocity.
Goreczny G.J., Ouderkirk-Pecone J.L., Olson E.C., Krendel M, & Turner C.E. (2016). Hic-5 Remodeling of the Stromal Matrix Promotes Breast Tumor Progression. Oncogene, 36(19), 2693-2703.
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5

Quantifying Fibronectin Matrix Clearance

Check if the same lab product or an alternative is used in the 5 most similar protocols
CAFs were plated onto 10μg/mL fibronectin coated glass coverslips and allowed to spread/clear the fibronectin matrix for 4 hours. Images of the cells were acquired using a Leica SP5 scanning confocal with a HCX PL APO 63×/1.40–0.60 OIL λ BL objective. Area of fibronectin fibers was quantified relative to the cell area using ImageJ.
Goreczny G.J., Ouderkirk-Pecone J.L., Olson E.C., Krendel M, & Turner C.E. (2016). Hic-5 Remodeling of the Stromal Matrix Promotes Breast Tumor Progression. Oncogene, 36(19), 2693-2703.
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