Laser capture microdissection and transcriptional analysis of sub-populations of the osteoblast lineage from undecalcified bone

Abstract

Transcriptional analysis of tissue samples is a useful and widely applied approach to provide insight into the molecular mechanisms engaged in response to phenotypic changes or external stimuli. We describe a method that overcomes the technical challenges associated with the application of Laser Capture Microdissection to undecalcified bone enabling us to collect high-quality bone tissue with maintained cellular morphology that is suitable for cryosectioning, fixation, and cutting. Using this method, we obtain samples enriched for specific cell types from the mature osteoblast lineage (osteoblasts, lining cells, i.e., quiescent osteoblasts, and osteocytes). RNA is well preserved and following extraction and amplification can be used as input to both low and high-throughput RNA analysis formats.