BAC RP23-201H14 was kindly provided by Dr. Cornelis Murre (University of California at San Diego). Position-specific 10-kb probes were generated by PCR using BAC templates with the primers listed in Supplemental Table 5 or described in Guo et al. (2011a (link)). FISH with 10-kb probes were performed as described in Guo et al. (2011a (link)) using a Nikon T200 microscope equipped with a 100× lens and motorized 100-μm Piezo Z-stage (Applied Scientific Instrumentation). Depending on the size of the nucleus, 30–40 serial optical sections spaced by 0.2 μm were acquired. The data sets were deconvolved using NIS-Elements software (Nikon). Volumes were measured using NIS-Elements software (Nikon). Statistical analyses of spatial distance measurements were carried out using a two-sample Kolmogorov-Smirnov test in R (Massey 1951 ).
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3D Spatial Imaging of Chromosomal Regions
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Fish
Lens
Microscope
Primers
Sections
T200
Corresponding Organization :
Other organizations : National Institute on Aging, Scripps Research Institute
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Variable analysis
independent variables
- Position-specific 10-kb probes generated by PCR using BAC templates
- Spatial distance measurements
- BAC RP23-201H14 provided by Dr. Cornelis Murre (University of California at San Diego)
- FISH performed as described in Guo et al. (2011a)
- Nikon T200 microscope equipped with a 100× lens and motorized 100-μm Piezo Z-stage (Applied Scientific Instrumentation)
- 30–40 serial optical sections spaced by 0.2 μm acquired
- Data sets deconvolved using NIS-Elements software (Nikon)
- Volumes measured using NIS-Elements software (Nikon)
- Statistical analyses of spatial distance measurements carried out using a two-sample Kolmogorov-Smirnov test in R (Massey 1951)
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