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Nasal passage aspirates, were collected as previously described, with sample from each neonate divided into two sections for detection of viral species by DFA and PCR, respectively.
AF647-conjugated streptavidin (4 μg/ml) and AF546-conjugated goat anti-hamster IgG (4 μg/ml) were added in TBS/1% fish gelatin onto the frozen sections for detection of CD11b+ cells and pdpn vessels in the dermis.
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We defined probes (described in Methods section) for detection of four MTBC species (M. tuberculosis, M. bovis, M. africanum and M. caprae) and 40 nontuberculous mycobacteria (NTM) species including M. abscessus, M. avium and M. intracellulare.
Gleno-humeral and acromio-clavicular joints were also assessed as following: dorsal transverse section through the infraspinous fossa laterally below the scapular spine and axillary longitudinal section, for detection of synovitis, synovial proliferation, and erosion of the humeral head; and ventral transverse section over the acromio-clavicular joint.
TUNEL assays were performed on lung tissue sections for the detection of apoptotic cells using an In situ Apoptosis Detection Kit (R&D Systems, Minneapolis, MN) following the manufacturer's recommendations.
He led us on a tour: there were separate sections for gene detection, gene amplification, and histological analysis.
Then sections for D2-40 detection were autoclaved for 7 min at 120°C in 10 mM citrate buffer solution (pH 6.0).
Tissue sections for fluorescent detection of SAHF were fixed in 3.7% neutral-buffered formalin overnight, dehydrated in ethanol and embedded in paraffin by standard pathological procedures.
It is known that CD4- and CD8-antigens are labile and is recommended to use freeze-tissue sections for their detection [ 29].
Co-localization studies of carbonyls and annexin V, a late marker in the apoptotic pathway, were not possible since sections for carbonyl detection had to be fixed in methacarn, which dissolves membrane phospholipids.
This improvement in detection accuracy comes at the expense of much greater computational cost, however (see Section 5 for detection accuracy and performance results).
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