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The spontaneous or maximum calcein release of target cells was identified by spontaneous release well (only target cells in complete medium) and maximum release well (only target cells in 2% Triton x-100-containing medium), respectively.
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Disruption of LUV by treatment with 1% Triton, causes total calcein release and a maximum increase in fluorescent intensity.
The maximum release and spontaneous release represent calcein release from the targets in the medium, with and without 2% Triton X-100, respectively.
The cell lysis was calculated as a percentage of the maximum released radioactivity: ((released radioactivity−spontaneous release)/(maximum released radioactivity−spontaneous release)) × 100%.
As shown in Fig. 3a, smaller alginate core size resulted in slower calcein release.
The calcein release profile was next examined for the three different alginate-PLGA microparticle formulations.
Based on the above analysis, alginate-PLGA microparticles containing 10 μm alginate cores demonstrated the highest calcein encapsulation efficiency (Additional file 2: Figure S2) and the slowest calcein release over 26 days.
Although microparticles containing 100 μm alginate cores (mag-mag) had the lowest encapsulation efficiency, their calcein release was the most rapid.
b Comparison of calcein release profiles of alginate-PLGA microparticles made through homo-mag and PLGA microparticles made through homo-mag.
Transmission electron microscopy (TEM), flow cytometry and calcein release assays revealed its excellent antimicrobial potency by inducing membrane permeation and disruption.
The rapid, concentration-dependent membrane depolarization, leakage of intracellular ATP and calcein release from PE/PG LUVs supported the membrane-lytic mechanism of action of the peptides.
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