A major obstacle for early warning of process failure at high FAN is that the additional ammonium/ammonia buffer covers up acidification.
Sections were mounted onto gelatin-coated microscope slides using aqueous mounting medium (3:1 glycerol/phosphate buffer), cover-slipped, and imaged with a Leica DM-RB upright microscope using Openlab software in the Biological Imaging Facility at Northwestern University.
The tube was inverted so the buffer covers the cap, vortexed at a medium speed for one minute and incubated at room temperature for 5 minutes.
Gravel roads with a 30 m buffer covered on average 5±0.5% (2 SE, range 2.9 7.2%) of the land area of the home ranges (n = 33).
Landscape composition (percentage of a given buffer covered by each habitat type) was calculated using hawth's tools (Beyer 2004) in arcgis 9.2.
Invert the tube and tap on the bench top several times to ensure the buffer covers the surface of the cap.
Liver tissue slices were placed individually into wells of 12-well plates containing 3 ml of ice-cold isolation buffer, covered and gently agitated (30 rpm) on ice using a metabolic shaker.
Some villages (Dembelela) had around 3% of surface within the 500 m buffer covered with very high and high-risk habitats, while it reached up to 60% in the vicinity of other villages (Sere, Tissi).
Tissues were lysed in 100 µl of Lysis Buffer covered with 600 µl of Denaturation oil at 95°C for 15 20 min followed by digestion with 1∶20 proteinase K (Ambion, Austin, TX, USA).
Mouse secondary biotinylated anti-human antibodies, specific for IgG1, -2, -3, and -4 and IgE isotypes (Sigma-Aldrich, Saint Louis, MO), were applied at a 1 100 dilution in blocking buffer, covered from light, and incubated overnight at 4°C.
Briefly, after rehydration in ethanol and fixation in 3.7%-buffered formaldehyde, sections were digested with proteinase solution (23 min at 37 °C), immersed in TdT labelling buffer, covered with the labelling reaction mix (37 min–1 h), immersed in streptavidin solution and dyed with DAB.
