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"Basically I think the data look like contamination," Dr. Manuelidis said, possibly stemming from "inadequately washed instruments".
Before analysis, samples from occipital lobe were placed on a hydrocarbon wax film (Parafilm M®, SPI Supplies Division, Structure Probe Inc., West Chester, PA, USA) and the gray matter (cortex) was separated from the white matter of the samples using acid washed instruments.
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At least two cycles of 5 minutes with PBS-Tween 0.05% followed by two cycles of 5 minutes with PBS in an automatic micro-plate washing instrument are recommended.
At least two cycles of 5 minutes with PBS-Tween 0.05% followed by two cycles of 5 minutes with PBS in an automatic micro-plate washing instrument are recommended to remove unattached protein.
Next, wash instruments to remove any debris or matter that might be left behind.
Before starting to clean your instruments, you must be sterile so you won't transfer any bacteria or germs to the instruments.You should wear a sterile gown while washing the instruments.
Wash your instruments and hands to avoid contamination.
A minimum number of 1000 events for each CSF staining was acquired for analysis with lyse-wash instrument settings, threshold FL-3 (PerCP) channel at 300 and the gate adjusted SSC versus FL-3.
After incubating for one hour at room temperature and washing repeatedly (at least two cycles of 5 minutes with PBS-Tween 0.05% followed by two cycles of 5 minutes with PBS in an automatic micro-plate washing instrument), a 100 µL volume of substrate solution (1-Step Ultra TMB-ELISA; Lot. 34028, Pierce®) was added to each well.
There was a report of plastic instruments being washed with Dial soap and reused.
Testing instruments were washed with double-distilled water and dried before use.
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