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Fd70s 100mg

Manufactured by Merck Group
Sourced in United States

The FD70S-100MG is a laboratory equipment product offered by Merck Group. It is a device designed for use in scientific and research environments. The core function of this product is to provide a precise and controlled environment for various laboratory processes. However, a detailed and unbiased description of the product's specific features and capabilities is not available at this time.

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6 protocols using fd70s 100mg

1

Retro-orbital Fluorescent Tracer Injection

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All the fluorescent molecules were diluted in saline solution (sodium chloride 9 g/L) and retro-orbitally injected in the blood, under isoflurane anesthesia (5%). In our hands, this route of administration is safer (more rapid, efficient and reproducible) than other routes of i.v. administration. Sham mice were injected with vehicle solution. Mice received only one injection with one fluorescent tracer. Phosphorothioate antisense oligonucleotide directed against angiopoietin-2 (Angpt2, named AS, GCG-TTA-GAC-ATG-TAG-GG, 6084.9 g/mol, Eurogentec) was coupled to 5-carboxyfluorescein (excitation: 492 nm, blue light; emission: 518 nm, green light) and injected (18 mg/Kg). Fluorescein isothiocyanate-dextrans D40, D70 and D150 (FD40-100MG, FD70S-100MG, FD150S-1G, respectively, Sigma-Aldrich, St. Louis, MI, USA) were solubilized in vehicle (2× g/100 mL to be injected retro-orbitally at 150 mg/Kg, near 200 µL/mouse). Fluorescein isothiocyanate-dextrans were maximally excited at 490 nm (blue) and maximally emitted at 525 nm (green).
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2

Measuring Pore Diffusivity in Microgel Scaffolds

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MAP gels were incubated with 100 µM 70 kDa dextran-FITC (FD70S-100MG, Sigma-Aldrich) solution in PBS or a 100 nM fluorescein solution in PBS. 20µL of microgels were pipetted and annealed in a 3-mm diameter PDMS well on a glass coverslip to form a MAP scaffold. Fluorescence recovery after photobleaching (FRAP) was conducted using a Leica TCS SP5 confocal microscope. A 20x dry objective and argon laser were used for bleaching and imaging. For pore diffusivity measurements, bleaching was performed with 30% laser power and 100% transmission, with imaging at 15% transmission to limit additional bleaching. For the single-phase bleaching measurements in non-porous hydrogel and PBS, 70% laser power and 100% transmission were used for bleaching, with 6% transmission used for imaging. After bleaching for 8 seconds, at least 50 images were taken with the interval of 390 ms (Figure S2). A circle of 100 μm diameter centered on the bleach spot was taken as the analysis region of interest (ROI) in all cases using ImageJ. The diffusivity was calculated via the approach of Soumpasis [44 (link)] (1):
D=.224w2t1/2
Where w is the ROI radius, t1/2 is the halftime calculated by fitting the mean intensity of the ROI in time to an exponential equation (2):
F(t)=a+b2t/t1/2
Where a and b were obtained from the fitted curve.
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3

Hydrogel Diffusivity Quantification

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8-mm disc gels were prepared as previously described in the rheology technique section. Gels were swollen in PBS overnight and placed between two slide glasses in PBS with 100 µM 70 kDa dextran-FITC (FD70S-100MG, Sigma-Aldrich). The fluorescent images of gels (FITC) were taken every day and the intensity profiles over time were used to calculate the diffusivity using Fick’s law.
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4

Fabrication of Porous MAP Scaffolds

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Fully swollen and equilibrated MAP building blocks (20 µL) were activated by with 5 U mL−1 FXIIIa (Sigma) and 1 U mL−1 thrombin, and the mixture was pipetted into a 3 mm diameter PDMS well on a glass coverslip and annealed in a humidified incubator at 37 °C for 1.5 h to form porous MAP scaffolds. Thereafter, the scaffolds were placed into HEPES buffer (pH 7.4) with 70 kDa dextran‐FITC (FD70S‐100MG, Sigma‐Aldrich) overnight to reach equilibrium. Samples were 3D imaged using a Leica TCS SP8 confocal microscope with 10× objective.
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5

Intracellular Dextran Tracking in Embryos

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Aliquots of 250-kDa dextran conjugated with FITC (FITC-250-kDa-dextran; Sigma-Aldrich, FD250S-100MG), 70-kDa dextran conjugated with FITC (FITC-70-kDa-dextran; Sigma-Aldrich, FD70S-100MG) and 3-5-kDa dextran conjugated with FITC (FITC-3-5-kDa-dextran; Sigma-Aldrich, FD4-100MG) were diluted to 12.5 mg/mL using ultrapure water (Thermo Scientific Barnstead Smart2Pure). Each solution was microinjected into the cytoplasm of both cells of 2-cell stage embryos, which had been preinjected with DNA-beads or control-beads at the 1-cell stage, using a piezo-activated manipulator with a narrow glass micropipette (1 μm diameter). DNA in the embryos was stained with 10 μg/mL Hoechst 33342 for 30 min. For the quantification of the fluorescence signals around DNA-beads, ImageJ software (http://rsb.info.nih.gov/ij/) was used. We first define the region of interest (ROI) to be quantified by converting the Hoechst 33342-stained image (DNA region) to the binary images using the threshold algorithm “Intermodes”. The FITC signals in the ROI and a cytoplasmic region of the same size were measured. The values were expressed as the ratio of the FITC signal of the ROI to that of the cytoplasmic region.
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6

Visualizing Diffusion Dynamics in Organoids

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To examine the spatiotemporal patterns of diffusion in OCTOPUS and hydrogel drops (Extended Data Fig. 2a-c and Fig. 5), we used 4-kDa FITC–dextran (FD70S-100MG, Sigma) as a fluorescent tracer for visualization. For this assay, the organoid culture medium was replaced with a FITC-dextran solution (50 μg ml−1 in PBS). Dextran diffusion was monitored and visualized using a laser scanning confocal microscope (LSM 800, Carl Zeiss). Time-lapse images were acquired for 120 min and processed using ZEN software (Zeiss) to measure temporal changes in fluorescence intensity at defined locations within the hydrogel scaffolds.
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