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Cell proliferation

Purpose

Cell proliferation defects are investigated using several assays: 

  • BrdU incorporation detects cells in S phase 
  • anti-Ki 67 immunohistochemistry detects cells which are in the cell cycle and distinguishes them from quiescent cells 
  • phosphorylated histone H3 marks mitotic cells and is detectable by immunohistochemistry.

Recommendations

When comparing cell proliferation between different tissues, all samples must be fixed and processed under the same conditions. At least, 3 mutant mice versus 3 littermate control mice are required for relevant results.

Apoptosis

Purpose

Apoptosis is investigated using two ways:

  • TUNEL (Terminal deoxynucleotidyl transferase-mediated dUTP Nick End Labeling); it represents the standard assay for detecting apoptotic cells on histological sections.
  • Cleaved Caspase-3 immunostainning

Recommendations

When comparing apoptosis between different tissues, all samples are fixed and processed under the same conditions. At least, 3 mutant mice versus 3 littermate control mice are required for relevant results.

Wound Healing

Purpose

Wound healing is a complex process requiring cell proliferation, extracellular matrix synthesis, inflammation, and contraction. The biological processes involved can reveal defects in immune response, nutrition, metabolism, or cell division. Wound healing analyses are performed on skin, which is a large and accessible organ, using full thickness excision method.

Recommendations

At least, 15 mutant mice versus 15 littermate control mice are required for relevant results.

Deliverables

Gross evaluation (healing rate and wound aspect evaluation) 

At each time point

  • Wound size and shape
  • Wound aspect
  • Edema formation and evolution
  • Presence of re-epithelialization
  • Curves of wounds surface along the time.

Histological evaluations

  • H & E staining (standard evaluation and fibroblast morphology)
  • Anti-neutrophil/granulocyte IHC staining
  • Anti-macrophage IHC staining
  • Anti-vascular endothelial IHC staining
  • Masson’s trichrome staining for fibroses 

Deliverables

Images in e-format or stained histological sections for quantitative analyses by the client.

BrdU incorporation
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