Laser Capture

Laser Microdissection and Pressure Catapulting (LMPC)

The PALM MicroBeam system from Zeiss allows users to microdissect individual cells or regions of tissue from their slides for downstream molecular analysis.

Why Use Laser Dissection?

  • Heterogeneous Nature of Tissue
    • Tumor Cells
    • Stroma
    • Lymphocytes

Heterogeneous tissue may confound molecular analysis because it is impossible to determine which cells contribute to which cellular constituent of a given lysate. LMPC gives you the ability to isolate specific cells of interest without contamination from surrounding cells.

The PALM MicroBeam System

  • Positioning Ablation Laser Micromanipulation (PALM)
  • Non-contact laser catapulting of tissue
  • Automation for efficient capture and pooling of large numbers of cells
  • Zeiss (Axiovert 200M) HotLInk Inverted scope offers a flexible platform for a variety of specimen types
  • Objective: 5x(non cutting), 10x, 20x, 40x, 63x (dry)
  • Fluorescence Filters: FITC,TxRd, Cy5

Specimen Requirements

  • OCT embedded frozen tissues
  • Paraffin embedded tissues
  • Cytology Preparations (i.e. blood smears, etc.)
  • Live Cell Cultures

Isolation of DNA

  • Mutation analysis
  • Whole Genome Amplification
  • Chromosome Spreads
    • Select individual chromosomes for sequencing of translocation break points

 

Beverly Emmanuel, CHOP


John Maris, CHOP

Isolation of RNA from Cryosections

  • Quantitative RT-PCR
  • cDNA array hybridization
  • Chromosome Spreads

 

Neil Rubenstein, UPENN

Isolation of Protein from Specific Cell Populations

  • Amplification is not possible with protein samples so larger samples are needed
  • Highly sensitive methods of protein analysis (SELDI,MALD) are now available.

 

Carsten Bonneman, CHOP

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