Materials and Methods
Immunohistochemistry. Immunohistochemistry was performed as
described (17). The primary anti-nestin (PharMingen) and antiphosphorylated
histone H3 (phospho-H3; Upstate Biotechnology,
Lake Placid, NY) Abs were detected with a Cy3-conjugated
secondary IgG (Jackson ImmunoResearch). High-power images
of mitotic neuroblasts were acquired and processed by using a
DeltaVision deconvolution microscope (Applied Precision, Seattle,
WA). For microtubule-associated protein 2 (MAP2) staining
of adult male brain, sections that had been hybridized with
X and Y chromosome paints and imaged were subsequently
stained with a monoclonalMAP2 Ab (Sigma) and an anti-mouse
Cy3 secondary IgG (Jackson ImmunoResearch). Cells identified
previously as aneuploid, based on fluorescence in situ hybridization
(FISH), were reimaged after MAP2 immunohistochemistry.
Images were prepared by using PHOTOSHOP (Adobe Systems,
Mountain View, CA).
Materials and MethodsImmunohistochemistry. Immunohistochemistry was performed asdescribed (17). The primary anti-nestin (PharMingen) and antiphosphorylatedhistone H3 (phospho-H3; Upstate Biotechnology,Lake Placid, NY) Abs were detected with a Cy3-conjugatedsecondary IgG (Jackson ImmunoResearch). High-power imagesof mitotic neuroblasts were acquired and processed by using aDeltaVision deconvolution microscope (Applied Precision, Seattle,WA). For microtubule-associated protein 2 (MAP2) stainingof adult male brain, sections that had been hybridized withX and Y chromosome paints and imaged were subsequentlystained with a monoclonalMAP2 Ab (Sigma) and an anti-mouseCy3 secondary IgG (Jackson ImmunoResearch). Cells identifiedpreviously as aneuploid, based on fluorescence in situ hybridization(FISH), were reimaged after MAP2 immunohistochemistry.Images were prepared by using PHOTOSHOP (Adobe Systems,Mountain View, CA).
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