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After laser ablation, individual apical or basal cells were collected manually and immediately washed to avoid possible contamination from the broken cells.
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From the sex separated and age pooled animals about 1000 antennae, 600 legs, 150 mouthparts (as piece of the head capsule anterior of the antennae), 50 heads (the whole head capsule excluding the antennae) and 20 bodies (excluding head and legs) were manually dissected and immediately transferred to ice cold RNA lysis buffer (Zymo Research, Irvine, USA).
Peak fractions were collected manually and tested immediately for lysozyme activity using the lysoplate technique [ 20].
The content of each intact cecum was extruded manually and frozen immediately until further use.
Eluting peaks were collected manually and neutralized immediately with 4 M acetic acid.
After a 90 min washout period, dialysate samples were collected every 6 min and immediately manually injected into a HTEC-500 HPLC unit (Eicom, Japan).
Manually, brachial artery blood pressure was measured before and immediately after each maximal exercise test.
Rats were euthanized using CO2 asphyxiation the day after behavioral testing, the brains removed, and the prefrontal cortex and hippocampus were manually dissected then immediately placed in liquid nitrogen and kept frozen until processed.
And immediately they did.
The stems were manually debarked immediately after felling.
The tubes were shaken manually at 10-min intervals for 1 h at room temperature and then centrifuged at 3,000 rpm for 10 min. Supernatants were transferred into sampling cups and immediately analyzed by observing the reaction with hexokinase.
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