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On a 16-core computer, an improvement of 14-fold in the analysis speed was noticed compared to sequential single-core approach, reaching an average time of 3 min per section.
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After being washed twice with PBS (5 min per wash), they were incubated with the StreptABComplex solution (1 ml per section) for 1 h at room temperature, washed twice with PBS (5 min per wash) and immersed into the substrate (300 ml PBS, 90 μl hydrogen peroxide and 2.5 ml 3,3-diaminobenzidine) for 3 min, and then rinsed with PBS and cold running tap water (5 min each).
After washing three times with PBS (5 min per wash), the sections were treated with 1% bovine serum albumin for 30 min at room temperature to block nonspecific reactions.
Solution access was limited to 30 min per session.
Then z-series scan (2 μm per section) was performed every 10 min for up to 4 h across each nuclear region.
During 60 min cells are incubated with Alexa 594-phalloidin (1 unit per section), dissolved in 0.2%FSG/PBS at 37°C.
Slides were incubated in 4 drops per section of Image-It FX Signal Enhancer (Invitrogen) for 30 min in a humidity chamber and then rinsed with PBS.
The sections were hybridized with [35S]-labeled oligonucleotide probes (7.5×105 dpm per section), washed, exposed to X-ray film (Kodak) for one week and developed (D-19, Kodak) for 6 min at 20°C.
Limited to 18 per section.
Limited to 20 per section.
Limited to 16 per section.
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CEO of Professional Science Editing for Scientists @ prosciediting.com