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Microscopic evaluation of the macrophage infiltration was done semiquantitatively using bright-field light microscopy coupled with digital camera.
In our study, we used tissue microarrays with core biopsies taken randomized from the tumor area and the quantification of lymphocytic infiltration was done by image analysis.
Infiltration was done with LR white resin (Ted Pella Inc., USA), first adding 1 : 1 LR White: Propylene Oxide for 2 h to the pellets and then stored overnight in 2 : 1 LR White: Propylene Oxide.
A time study of oil and starch content as well as RT-qPCR and photosynthesis measurements in leaves expressing Arabidopsis WRI1 between one to five days after infiltration was done to further characterize and complement the transcriptional transitions observed at five days after infiltration.
Microwave resin infiltration was done using a 1 1 mix of Spurr's resin to ethanol at 250 W at 20 °C for 3 min in a vacuum; two additional infiltrations were done in pure Spurr's resin, 25 W at 20 °C for 3 min in a vacuum.
An assessment of neutrophil infiltration was done by a conventional H&E staining and in our material showed an increased numbers of neutrophils from day 6 of exposure to DSS. Specific immunohistochemical staining was not done in this study as it has been done by others.
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All infiltrations were done in plastic syringes.
All infiltrations were done using 50-mL syringes and 7-cm long 20-G needles.
Infiltration of the samples was done by placing them in a mixture of absolute acetone and the final spurr resin mixture (1 1) for 1 h at room temperature.
Fabrics were layered and compacted using a thermoplastic veil while infiltration of the preforms was done using the vacuum assisted process.
The subsequent infiltration of PEG 1500 was done at 50 °C as follows, 30 min 100%% EtOH, 60 min 25 % PEG in EtOH, 60 min 50 % PEG in EtOH, 60 min 75%% PEG in EtOH, 60 min 100%% PEG in EtOH.
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