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Pnp tr tips

Manufactured by NanoWorld

PNP-TR tips are a type of disposable pipette tips designed for use with various laboratory equipment. The core function of these tips is to serve as a disposable interface between the pipette and the sample, facilitating accurate and reliable liquid transfer during various laboratory procedures.

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2 protocols using pnp tr tips

1

Measuring FLNC-Knockdown Cardiomyocyte Stiffness

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Control and FLNC-knockdown NMVCMs were prepared as described in Section 4.7 and cultured on laminin-coated micropatterned PDMS [43 (link)]. Atomic force microscopy (AFM) was performed on an MFP-3D Bio Atomic Force Microscope (Oxford Instruments) mounted in a Ti-U fluorescent inverted scope (Nikon Instruments, Melville, NY) and used Asylum Research 13, Igor Pro 6.34A software. Nanoworld PNP-TR tips were calibrated for their spring constant using the thermal noise method and used for probing the FLNC-knockdown NMVCMs and their controls. A trigger force of 2 nN, an approach Velocity constant of at 2 µm/s, and a force-distance of 6 µm were used to generate a force map with 12 points across 5 µm2 [45 ]. Asylum Research 13, Igor Pro 6.34A software was used to calculate the slope of the deflection-indentation relationship using the Hertz equation (Hertz, H. Ueber den kontakt elastischer koerper. J. fuer die Reine Angew. Math.92, 156 (1881)) and taken as the Young’s Modulus, or stiffness, (in kPa) of the cell membrane/cortical cytoskeleton. Poor fits to the deflection-indentation were excluded and the average Young’s modulus was calculated from the remaining force map points. One cell was measured per force map.
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2

Probing Nuclear Mechanics via AFM

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For atomic force microscopy, isolated nuclei in PBS were spun (500g, 3 minutes) on to 12mm coverslips coated with Poly-D-Lysine (1 μg/μl was used to coat coverslips for 5 minutes, then rinsed with purified water and left to dry overnight). Coverslips were transferred to a glass slide, secured with vacuum grease, and covered in a PBS droplet for AFM. Indentation experiments were performed on an MFP-3D Bio Atomic Force Microscope (Oxford Instruments) mounted in a Ti-U fluorescent inverted scope (Nikon Instruments, Melville, NY) and used Asylum Research 13, Igor Pro 6.34A software. Nanoworld PNP-TR tips were calibrated for their spring constant using the thermal noise method and used for probing isolated nuclei. A trigger force of 2nN, an approach Velocity constant of at 2 μm/s and a force distance of 6 μm were used to generate a force map with 12 points across 2μm2. Hand4.2-Gal4 was used to drive expression of GFP, and thus only GFP-positive nuclei were selected for indentation. The software was used to calculate the Young’s modulus using the Hertz equation89 . Any poor fits to the indentation curve were excluded. Then, the average Young’s modulus was calculated from the force map.
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