Transcriptome Profiling of Murine Spinal Neurulation Using Laser Capture Microdissection and High-Density Oligonucleotide Microarrays
Neurulation is a critical process in the formation of the central nervous system during embryonic development. Closure of the neural tube is driven by forces that originate from both the neuroepithelium and the surrounding tissues. In this chapter, we describe the use of laser capture microdissection to isolate and separately collect cells from the neuroepithelium and the underlying mesenchyme. We provide protocols for processing of samples for downstream comparison of the transcriptomes of two cell populations using high-density oligonucleotide microarrays, with an emphasis on important technical issues that are to be borne in mind when carrying out these experiments.
Key wordsLaser capture microdissection Mammalian spinal neurulation Transcriptomic analysis High-density oligonucleotide microarray
We thank Associate Professors Ming Teh and Wei-Yi Ong for kindly allowing us to use their laser capture microdissection systems, and their colleagues for help with operating the instruments. We are also grateful to Ms Song-Lin Bay for her excellent assistance in preparing the diagrams for this chapter. The work was supported by Grant R-181-000-095-112 from the Academic Research Fund, Ministry of Education, Singapore (G.W.Y.). S.C. is the recipient of a graduate research scholarship from the National University of Singapore.
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