HiMolA Platform

The Molecular Histology (HiMolA) platform is one of the 6 technical platforms hosted by the RMeS unit and based exclusively at the University of Angers.

The HiMolA platform regroups three main activities:

1) Investigation of bone microarchitecture and histomorphometry

2) Investigation of bone material properties

3) Investigation of bone biomechanics

1. Investigation of bone microarchitecture and histomorphometry

HiMolA offers its service in conventional bone histomorphometry including bone preparation (calcified and decalcified) and embedding, sectioning, and staining. This platform routinely performed several stainings for the identification of bone marrow cells (toluidine blue-borax), osteoclasts (tartrate-resistant acid phosphatase), osteoblasts (alkaline phosphatase), osteoid tissue (Goldner trichrome) or metal accumulation (Perl’s, solochrome azurine). Static morphometric parameters, indicative of the structure and activity at the time-point of the collection of the bone specimen, can be measured according to the American Society for Bone and Mineral Research Guidelines. Dynamic parameters, indicative of the amount of bone formation between two given time points, can also be determined. In addition, the platform has also expertise in performing molecular detection by immunohistochemistry of bone samples and analyses of such samples.

The platform is also specialized in X-ray imaging of calcified tissue by micro/nanoCT  and propose several solutions ranging from in-vivo investigation of bone microarchitecture in small animals to ex-vivo analysis of specimen in three dimensions (3D). These solutions are useful in determining volumetric bone mineral density and to characterize the microarchitecture of bone specimen.

The platform is equipped with :

• Leica Polycut
• Struers Accutom-50
• Struers Labotom-3
• Exakt saw
• Exakt polishing machine
• Struers Tegramin-30
• Fume hood
• Faxitron LX-60 cabinet
• Skyscan 1272 ex vivo microCT
• GE Phoenix nanotom s nanoCT

2. Investigation of bone material properties

The bone matrix is composed mainly of three components represented by a mineral phase composed of poorly crystalline carbonated hydroxyapatite, an organic phase composed mostly of type I collagen and water. The HiMolA platform has developed over years a strong expertise in the assessment of bone material properties, i.e. the properties of the component of the bone matrix. Indeed, this platform propose a range of dedicated services to study (i) the mineralization pattern of bone samples and other mineralized tissues by quantitative backscattered electron imaging (qBEI), (ii) the maturity and size of mineral crystallites and organic component of the mineralized tissues by Fourier transform infrared microspectroscopy or imaging (FTIRM, FTIRI) or by Raman microspectroscopy. The HiMolA platform also holds human reference biopsy that made possible comparison between healthy and pathological bone samples.

This platform holds or has access to the following equipments:

• Zeiss Evo LS10 scanning electron microscopy equipped with a picoamperemeter and a faraday cage
• Bruker Hyperion 3000 infrared microscope, with MCT and FPA detectors, coupled to a Vertex 70 infrared spectrophotometer
• Renishaw InVia confocal Raman microspectrophotometer

3. Investigation of bone biomechanics

Investigation of bone biomechanics is crucial in order to evaluate fracture resistance and eventually bone fragility. The HiMolA platform offers a series of dedicated service to study bone biomechanics either at the organ (three-point bending, compression test) or tissue level (nanoindentation). These investigations, in addition to the other service provided by the HiMolA platform, allows for deciphering the biomechanical mechanisms responsible of the bone phenotype. The HiMolA platform has also expertise for the preparation and evaluation of bone samples from mouse to human. This platform is equipped with:

• Instron 5972 test machine
• Instron Bluehill 3 software
• CSM NHT2 nanoindentation station