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Many presenters adopt the quickest, least creative method and use the poster as a mounting medium for their paper.
UltraCruz® mounting medium for fluorescence studies with DAPI was supplied by Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).
The cells were fixed with 4% formalin for 10min and washed with PBS and mounted with the DAPI mounting medium for nuclear staining.
DAPI (1 ng/mL) was added to Vectashield (Vector Labs) as mounting medium for visualization of DNA.
The sections were dehydrated in a 70% 96% 100%0% alcohol series, washed in xylene for 2 minutes, and then mounted with DPX mounting medium for histology (Sigma-Aldrich).
For confocal analysis, embryos were stained with DAPI-Vectashield and TRITC-phalloidin mounting medium for 20 mins, mounted in 1∶1 Glycerol PBS and analysed with a Leica DM IRB upright confocal microscope.
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Sections were again rinsed and coverslips mounted with Prolong Gold mounting medium (Life Technologies) for microscopic analysis.
The slides were air dried and mounted with an aqueous mounting medium made for fluorescence (Gel Mount), with DAPI (4′,6-diamidino-2-phenylindole) (VectaShield HL1200).
Cells were fixed in 4% paraformaldehyde for 30 min at RT and mounted in Vectashield ® hard mounting medium containing DAPI Vector Laboratoriess) for analysis by confocal microscopy (Zeiss LSM 510).
For fluorescent staining, cryostat sections were rinsed in PBS to remove mounting medium and blocked for 1 h at room temperature in 5% normal donkey or goat serum/0.02% Triton X-100/PBS.
The mean OD was determined by rounding off the stained structure of interest (TNC) and subtracting the OD of the background (slide, mounting medium and coverslip) for each section, considering a total of 12 sections per animal.
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