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"If we use two parallel surfaces, it is very hard to push to nanometer scale without some parts touching each other," Chen explains, but by using a bead there is just a single point of near-contact, which is much easier to maintain.
Liver tissue (≈3 mg) was homogenized by using a bead mill.
To assess TGase activity in articular homogenates, tissue lysates were prepared from freshly excised control and CIA rats (21 days after induction) synovial membranes, homogenized by using a bead mill (Retsch, Newtown, PA, USA) in TGase reaction buffer (50 m M Tris, pH 7.5; 150 m M NaCl; 5 m M CaCl2; Complete (Roche)).
Brain and liver extracts for enzyme assays were prepared from ∼50 mg frozen tissue under native conditions [lysis buffer: 50 mmol/L potassium phosphate, 0.3 mmol/L EDTA, supplemented with protease (Roche) and phosphatase inhibitors (Thermo Scientific)] by using a bead mill and cleared by centrifugation.
DNA was isolated from frozen stool specimens by using a bead beater with 3-mm diameter solid glass beads (Sigma-Aldrich, St . Louis MO, USA) and, subsequently, with 0.1-mm zirconium beads (BioSpec Products, Inc., Bartlesville, OK, USA) to disrupt cells.
The two easiest ways to finish are by either tying the ends or by using a bead, button, or knot clasp.
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Wagner et al. performed the most detailed analysis so far by using a bead-based array that included a larger number of citrullinated antigen (n = 16) in a small cohort of patients with RA, disease controls and healthy controls (n = 78) [ 10].
Tissue pools were homogenized by using a Precellys bead mill homogenizer (Bertin Technologies, Ann Arbor, MI, USA), and DNA was extracted by using the Wizard Genomic DNA Purification Kit (Promega, Southampton, UK) according to manufacturer's instructions.
Serum IL-27 (EMD Millipore Corporation, Billerica, MA, USA) and procalcitonin (Bio-Rad, Hercules, CA, USA) protein concentrations were measured by using a magnetic bead multiplex platform and a Luminex 100/200 System (Luminex Corporation, Austin, TX, USA), according the manufacturers' specifications.
A small section of each tissue was collected, weighed, and homogenized by using a metal bead in 1 mL of DMEM containing 100 U penicillin and 100 μg/mL streptomycin.
We achieved ∼1.1 nm for the target registration error for mapping transformation between the two color probes (Supporting Information notes and Supporting Information Figure 2b and c), three times better than that obtained by using a fluorescent bead and moving the microscope stage, or by using nonuniform nanoholes.
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