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Development and Growth disorders are usually investigated for lethal or subviable mutations; this occurs for 24% and 10% of the genes, respectively (IMPC data). These genes are of special interest for both developmental biologists and clinicians as they are potential candidates for human developmental disorders and rare genetic diseases.

Suggested primary analysis

See our Anatomopathology dedicated page to explore all our available tests.

Purpose

Cell death by apoptosis can be investigated in ex vivo sections by TUNEL staining and/or cleaved caspase 3 immunohistochemistry.  

Purpose

Tissue cell proliferation markers provide information on cell cycle progression with BrdU as an indicator of cells having passed through S-phase, phospho-H3 for cells in M-phase, and Ki67 for actively cycling cells.

Purpose

To better characterize the primary defects of the mutants, vascular (PECAM), neural (neruofilament), endodermal (HNF3b) markers are proposed. Other marker stainings can be developed on demand.

Embryo analysis

Purpose

Three dimensional reconstruction of embryo slices provides the definition of histological staining with the completeness of 3-D imaging.

Purpose

Rapid three dimensional imaging of density-contrasted tissues in the embryo using X-ray transmission and computed tomography.

Purpose

Sampling of embryos for viability at times throughout pregnancy with focus on development of particular organs between e9.5 and e18.5.

Purpose

Embryonic growth restriction can be indicative of placental abnormalities. Macroscopical and routine histological analysis (hematoxylin & eosin) are performed to evaluate placental defects. 

X-Ray imaging

Suggested secondary analysis

Purpose

The measurement of various blood metabolites, ions, and enzymes provides a primary screen for  function of major metabolic organs such as kidney, liver, gastrointestinal tract, as well as for lipid and glucose homeostasis. 

A panel of parameters is proposed, including:

  • Electrolytes and ions: Sodium (Na), Potassium (K), Chloride (Cl), Calcium (Ca), Magnesium (Mg), Phosphate (PO4), iron (Fe), and bicarbonate (CO2).
  • Enzymatic activities: Alpha-amylase, aspartate amino transaminase (AST), alanine amino transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), lipase, creatine kinase (CK).
  • Metabolites: glucose, urea, creatinine, uric acid, total proteins, albumin, total bilirubin, triglycerides, free fatty acids, glycerol, total, HDL and LDL cholesterol, fructosamine, ketone bodies (B-HBA), transferrin, ferritin.

Biochem_Clinical Chemistry graph.jpg

Selected plasma chemistry data from female mice (N=20 per line) of common laboratory strains at different ages (Champy et al. Mammalian Genome 2008).  Error bars reflect ± SEM.

Equipment

These tests are performed with an AU-480 automated laboratory work station (Beckman Coulter France SAS, Villepinte, France).

Strain background references

Champy MF, Selloum M, Zeitler V, Caradec C, Jung B, Rousseau S, Pouilly L, Sorg T, Auwerx J. Genetic background determines metabolic phenotypes in the mouse. Mamm Genome. 2008 May;19(5):318-31. Epub 2008 Apr 5.

Purpose

Body composition for fat, lean and free body fluid is evaluated on conscious mice by quantitative nuclear magnetic resonance on Minispec analyzer.

Metabo_Body composition by qNMR graph

Body composition analysis of male mice from different common laboratory strains after provision of chow diet (CD) or 10 weeks provision of high-fat diet (HFD), including complete body weight, fat tissue content, and lean tissue content.  Dramatic increases in fat tissue is seen upon HFD-diet treatment for most strains excepting BALB/c, consistent with what has previously been reported. 

Equipment

Metabo_Body composition Equipement picture

Body composition is evaluated by Quantitative Nuclear Magnetic Resonance (qNMR) using Minispec+ analyzer (Bruker BioSpin S.A.S., Wissembourg, France)

Purpose

First-line hematological/immunological analysis of blood or tissue infiltrates for different leukocyte lineages. May be targeted to examine particular immune lineage subclasses.

Exemples:

Thymus: 

  • T cells of ab lineage: CD4/CD8/TCRab 
  • T cells of gd lineage: CD4/CD8/TCRgd 
  • NKT cells: NK1.1/CD3 

Spleen: 

  • B cells: CD19/IgM/IgD/CD80 
  • B cells: CD19/IgM/CD2/CD23 
  • NK cells and NKT cells: DX5/NK1.1/CD3 
  • Macrophages and neutrophils: F4-80/Gr-1/Mac-1 
  • T cells: CD4/CD8/CD3/CD44

Blood:

  • B cell: CD19/mature B cells IgD
  • Granulocyte
  • Monocyte
  • NK cell 
  • T cell : CD3/CD4/CD8
  • Treg cell : CD25

Please see our Immunology dedicated page for more detailed tests.

Equipment

FACS analysis are performed on a LSR II (BD diagnostics, Le pont de Claix, France).

See our Gene expression analysis dedicated page to explore our different related tests.

Food challenge

Purpose

With several months notice, we can procure and provide a variety of different restriction diets for nutritional analyses in conjuction with our other analyses.

Ultrasounds imaging

X-Ray imaging

Purpose

The X-Ray system gives very precise images of the skeletton. In DXA mode it automatically calculates BMD, BMC, and lean and fat mass percentages.

Strain background references

Champy MF, Selloum M, Zeitler V, Caradec C, Jung B, Rousseau S, Pouilly L, Sorg T, Auwerx J. 
Genetic background determines metabolic phenotypes in the mouse. 
Mamm Genome. 2008 May;19(5):318-31. Epub 2008 Apr 5.

Equipment 

Imaging_Bone mineral density equipment DEXA

pDEXA Sabre (Norland)

Sample data

Imaging_Bone mineral density image DEXA

Recommendations

  • For performance in conjunction with X-ray analysis.
  • 8 mice per group are recommended for reliable data analysis.

Models and Challenges

Purpose

With several months notice, we can procure and provide a variety of different restriction diets for nutritional analyses in conjuction with our other analyses.