Laser Capture Microscopy
Asterand Bio scientists have expert training in LCM using their in-house Molecular Machines CellCut™ system for the isolation of specific regions/cells of complex tissues. Utilizing hematoxylin directed and validated LCM in combination with qRT-PCR, we are able to quantify mRNA expression in accurately dissected tissue areas of interest.
The Molecular Machines CellCut™ system uses fixed UV-laser with high pulse rate but low power. This results in ultra precise cutting (0.3 mm line width) and minimal sample damage.
Morphological region specific gene expression
Morphological specific gene expression following LCM of kidney regions. Glomerulus, ducts and tubules were captured via LCM and qRT-PCR for region-specific genes was preformed.
Sections of normal kidney cortex were subject to LCM for specific morphological regions: glomerulus, ducts and tubules.
From the resulting sub-dissected samples, qRT-PCR for region-specific genes shows clear distinction of expected genes in the expected regions.
- Renal cortex (n=3), glomeruli vs tubules: measurement of POD, AQP1 and CK7 mRNAs
- Podocin mRNA high in glomeruli, low in ducts
- Cytokeratin 7 high in ductal / tubule regions, low in glomeruli
- Aquaporin mRNA low in glomeruli, higher in ductal / tubules regions.
- Accurate correlation with separate IHC for these markers in these regions
To validate reproducibility of the LCM system we cut standard sized samples of liver parenchyma and looking at the expression of GAPDH (housekeeping gene) – data not shown here.
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