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Several tissue sections were analyzed for each animal and the results are expressed as mean ± SD values.
To analyze such small features, however, sufficient numbers of ROIs must be captured, and this may necessitate pooling photo-tagged material from several tissue sections.
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After several washes, tissue sections were incubated with the secondary antibody, Alexa Fluor 568 goat anti-rabbit IgG (1∶1000, Molecular Probes) for 1 hr at room temperature.
After several washes, tissue sections were incubated with the secondary antibody, anti-mouse rhodamine red (1∶1000, Molecular Probes) for 1 hr at room temperature.
After several washes, tissue sections were incubated with the secondary antibody, Alexa Fluor 568 goat anti-mouse IgG or donkey anti-rabbit IgG (1∶800; Molecular Probes) for 1 hr at room temperature.
Unfortunately, several separate tissue sections at different depths of the tissue block should have been examined to assess the effects FFPE has on reproducibility.
Briefly, only cancerous regions were microdissected using a sterile 26-gauge needle from several serial tissue sections (20 μm thickness) under visualisation with an inverted microscope (Nikon 66906, Tokyo, Japan).
Areas with weak staining were observed in several of the tissue sections.
Previous studies have used data from several non-serial tissue sections viewed by light microscopy to extrapolate a volume and have assumed that MLBs are random in distribution.
RNA was also extracted from several paraffin-embedded tissue sections from HD patients, using the Paraffin Block RNA Isolation kit (Ambion Inc., The RNA Company, Austin TX, USA) and RT PCR followed by nested PCR was performed with 10 μl of extracted RNA as described above.
Next, the pattern of KLF6 protein was analyzed by immunohistochemistry assays in several normal and tumor tissue sections derived from paraffin-embedded samples spotted in an array.
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CEO of Professional Science Editing for Scientists @ prosciediting.com