HE染色及免疫组化英文步骤组织名称were immersed in 4% paraformaldehyde for 4h,and transfered to 70% ethanol.Individual lobes of 组织名称biopsy material were placed in processing cassettes, dehydrated through a serial alcohol gradient, and embedded in paraffin wax blocks. Beforeimmunostaining, ...
组织名称were immersed in 4% paraformaldehyde for 4h,and transfered to 70% ethanol.Individual lobes of 组织名称biopsy material were placed in processing cassettes, dehydrated through a serial alcohol gradient, and embedded in paraffin wax blocks. Beforeimmunostaining, 5-um-thick lung tissue sections were dewaxed in xylene, rehydrated through decreasing concentrations of ethanol, and washed in PBS.And then stained with hematoxylin and eosin (H&E). After staining, sections were dehydrated through increasing concentrations of ethanol and xylene.
Immunohistochemistry for CCR7,CD206,and F4/80.Antigens were unmasked by microwaving sections in 10mmol/L citrate buffer, pH 6.0 (15 minutes), and immunostaining was undertaken using the avidinbiotinylated enzyme complex method with antibodies against CCR7at a concentration of 1ug/ml, CD206 at a concentration of 1 ug/ml, F4/80 at a concentration of 1 ug/ml, and equivalentconcentrations of polyclonal nonimmune IgG controls. After incubation with the appropriate biotin-conjugated secondary antibody and subsequently with streptavidin solution,color development was performed using 3,3 -diaminobenzidinetetrahydrochloride
(Vector Laboratories) as a chromogen.Sections were counterstained using Gill-2 hematoxylin (Thermo-Shandon, Pittsburgh, PA). After staining, sections were dehydrated through increasing concentrations of ethanol and xylene.
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