Laser capture microdissection and laser pressure catapulting as tools to study gene expression in individual cells of a complex tissue.
Laser capture microdissection (LCM) method allows the selection of individual or clustered cells from intact tissues. LCM enables to pick cells from tissues that are difficult to study individually, to sort the anatomical complexity of tissues, and to make the cells available for molecular analyses. This technology provides an opportunity to uncover the molecular control of cellular fate in the natural microenvironment. It is a difficult task to obtain cells from skeletal tissues, such as cartilage, periost, bone, and muscle, that are structured together and do not exist as individual organs. LCM allows isolation of desired cells from the native tissue environment for the analysis of gene expression. We earlier described the selection of cells from skeletal tissues that were analyzed for expression of transcription factors, receptors for cytokines, nuclear receptors, and functional genes such as alkaline phosphatase and structural proteins. Current results acquired using the LCM technology demonstrate expression of known genes that are in agreement with their reported in vivo and in vitro function in skeletal cells. The obtained knowledge will provide molecular information in the context of the cell and tissue biology. Such analysis will enable a reliable interpretation of function of known and novel genes expression in the skeletal tissues under various physiological conditions.
Dafna
Benayahu
B
Department of Cell and Developmental Biology, Sackler School of Medicine, Tel-Aviv University, Israel.
Rina
Socher
S
Irena
Shur
S
D000818:Animals; D001842:Bone and Bones / Q000166:cytology; D002356:Cartilage / Q000166:cytology; D015870:Gene Expression; D005786:Gene Expression Regulation; D007834:Lasers; D051379:Mice; D042282:Microdissection / Q000379:methods*; D012313:RNA / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013048:Specimen Handling; D016707:Tissue Fixation
D012313:RNA