Histology and Correlative Microscopy Core

Preparation laboratory

The HCMC provides comprehensive facilities for the preparation of samples for light and electron microscopy and for molecular and inorganic elemental diagnostics. Tissue preparation and histology is specialized on retaining the mineral in calcified tissues, such as in bones, teeth, fossils, grafting materials, and calcium phosphate scaffoldings, and for the preparation of special materials such as metal, ceramics, and implants. Specimen embedding is performed using either epoxy or acrylate-based resins cured at ambient or low temperatures. Thin section preparation includes both manual and automated grinding, polishing, and cutting to provide sections typically 100 micrometers thick, plus or minus. Plastic embedded thin sections may also be cut with a microtome to produce thicknesses in the range of 5-15 micrometers.

• Buehler grinding and polishing and undecalcified tissue preparation
  
• Buehler thin sectioning
• Exakt preparation and thin sectioning，grinding and polishing system
• Leica RM2265 fully motorized rotary microtome  

Gigapixel imaging in 2D

The HCMC specializes in providing high resolution gigapixel images of histological thin sections and block surfaces typically within 5-20 minutes. This applies variously to Leica and Zeiss transmitted and/or reflected bright field, linear or circularly polarized light, phase contrast, circularly differential interference contrast, and fluorescence microscopy. From images of say 50k x 50k pixel dimension, any field of microscopic interest may be extracted from the macro image. Matching LM and SEM fields of view is also provided by Zeiss correlative microscopy.

• Zeiss Axiolmager M1m + Correlative Microscopy and gigapixel montaging workstation
• Leica-Aperio GL ScanScope
• Leica DMRXE Universal Microscope + Objective Imaging gigapixel montaging workstation
• Epson V800 Photo scanner

Confocal microscopy

HCMC confocal microscopy includes conventional laser imaging, but also has superior refection imaging white light confocal microscopes, including a portable model.

• Technical Instrument K2-S BIO Confocal Microscope
• Technical Instrument K2-S BIO Portable Confocal Microscope
• Zeiss AxioVert 200 + Perkin-Elmer UltraView ERS 3E Laser Confocal Microscope

Molecular and elemental diagnostics

HCMC is the only laboratory in the world to provide simultaneous and absolute quantitative elemental composition of liquids and solids across the full spectrum of inorganic elements the periodic table (71 of 92 elements). Transmission and reflection Fourier Transform Infrared spectroscopy is also available.

• SPECTRO MS Laser Ablation Inductively Coupled Plasma Mass Spectrometer
• Perkin Elmer Spectrum 100 FTIR

3D visualization, Imaging, and measurement

The HCMC is first in the world to have both transmitted and reflected analog real time 3D workstations, as well as fully digital real time 3D microscopy employing Sony gaming glasses and a 3D monitor. In addition, a 3D measurement system operating in a 10cm cubic volume with approximately 2 micrometer accuracy, a surface metrology system with an ultimate 0.9 micron resolution, and conventional stereo zoom microscopy are available.

• EDGE 3D Direct View 3D Microscope
• Edge R400 Realtime 3D Microscope + Syncroscopy Z-stacking 3D workstation
• Edge H160 Realtime 3D microscope + PhaseView profilometry and Syncroscopy Z-stacking 3D workstation
• Mitutoyo custom non-contact 3D measurement system
• Zeiss Axiolmager M1m + PhaseView profilometry
• Leica MZ-APO Stereo Microscope

Electron Microscopy

The HCMC has both Hitachi and Zeiss scanning electron microscopes operating in both high vacuum and variable pressure modes. The field emission scanning electron microscope has a wide ranging gigapixel imaging, 3D metrology, and correlative microscopy imaging capability. Both SEMs include density-dependent backscattered electron imaging in the SEM (BSE-SEM) and Bruker energy dispersive spectroscopy (EDS) elemental analysis.

• Zeiss Gemini 300 FE-SEM + Correlative Microscopy, Atlas montaging, 3D profilometry, and Bruker EDS elemental analysis
  
• Hitachi S3500N SEM
• SEM coating workstation

Zeiss Gemini 300

field-emission scanning electron microscope with Bruker EDS

Features

• SE, BSE, In Lens SE, In Lens voltage-gated BSE Variable Pressure Imaging
• Zeiss Correlative Microscopy
• Zeiss Atlas gigapixel montaging
• 3D visualization and quantitative measurement system
• Bruker Energy Dispersive X-Ray Spectroscopy

Image Analysis

The BIOQUANT OSTEO software enables cutting-edge bone biology research in animal models and human biopsy preparations. It supports both high-throughput automation for ASBMR standard bone histomorphometry in digital histology and microCT. Primary applications include skeletal phenotyping, human histmorphometry, cancer metastasis, cortical bone structure, osseointegration, chondrocyte proliferation, arthritis, and sarcopenia.

• BIOQUANT Osteo Measure histomorphometry workstation
• Leica DM5000 B + Leica Quantimet v.3 image processing workstation