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Samples were incubated in staining mix for 15 min prior to flow cytometry.
To evaluate plant cell death, pathogen-infected leaves were treated with lactophenol trypan blue staining mix [ 31].
After 2 washes with TBS for 2 min each time, the slides were incubated with DAB staining mix for 5 min and then washed with water.
Senescent cells were stained by adding 1 ml of the staining mix to each well.
To detect cell death, sections were then incubated with staining mix (in situ cell death detection kit, TMR red, Roche) according to manufacturer's instructions.
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While waiting for incubation, staining mixture was prepared by mixing 1 mL of staining solution 10×, 125 μL of reagent B, 125 μL of reagent C, 250 μL of X-gal solution (pre-warmed for 1 h at 37°C) and 8.5 mL of miliQ water.
If soap and water do not remove the stains, mix up a solution of Oxy Clean® and place the stained area in the solution to soak until the stain disappears.
Fienberg H, Simonds EF, Fantl WJ, Nolan GP, Bodenmiller B. Description: Barcoding cells from different samples with specific isotope combinations allows mixing, staining and measuring multiple samples together in one go, thus eliminating multiple sources of sample-to-sample variability.
Briefly, cells were washed with HBSS, fixed using a fixative solution for 10 15 minutes, again washed with PBS, and then incubated with Staining Solution Mix overnight at 37°C.
Microscopic observation of cells following treatment with the CDP mix and staining with DAPI showed that while HeLa cells treated with DMSO solvent alone did not exhibit nuclear DNA fragmentation), HeLa cells did exhibit nuclear DNA fragmentation after treatment with the CDP mix at a concentration of 10 mg/mL for 24 h); in addition, apoptotic bodies were clearly visible in cells treated with CDPs).
T-cells were stained with BD Fast-Immune mix of CD3/CD4/CD69.
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