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   Service List::
NDT501: Stereology-based morphological analysis of CNS (volumetric measurement, estimation of cell populations/cell counts, density and single neuron quantitation)

Unbiased stereology has been considered by modern neuroscientists as the preferred method to measure or quantify neuronal morphology of the CNS.  This is particularly important in identification of subtle yet important alterations in the morphology of the CNS based on disturbances during development or in neuropsychiatric and neurodegenerative diseases.

The core focus of NDT501 service is designed for accurate measurements of neuronal elements (including size, shape and number) obtained in an unbiased way by implementation of proven software packages and algorithms that include standard geometric overlays, mathematical formulae, counting rules, and systematic random sampling methods.  Using this method, the obtained results will be accurate, efficient and more reliable than other ad hoc quantitative analyses.

With computerized systematic random-sampling layout and stabilized stage control of spatial (x-y-z) coordinates on every serial section of Region of Interest (ROIs) via Zeiss digital microscopic platform, we are able to quantify properties of 3D objects from a stack of 2D serial sections through an object (such as microscopic slides of a solid specimen). Our clients will expect to receive most accurate and reliable morphometric results, namely, a “3D” spatial digital profile of ROIs with a series of digitized data, ready for subsequent digital archiving and data/quality assurance.  

NDT501 service is capable of measuring brain volume and cell population estimate based on the principle of stereology:

I. 3D volumetric measurement of hippocampus

II. Estimation of cell populations / cell counts

3D volumetric analysis of mouse hippocampal subfields

3D distribution of BrdU+ cells in mouse dentate gyrus (*Cross-lines between dentate gyri were automatically generated by the software for maintaining stabilization of spatial coordinates)

Based on the principles of the modern stereology, NDT501 service is used to systematically quantify neuron morphology, including dendritic analysis and spine counts/density along the anterior-to-posterior axis of Region of Interest (ROI) of the CNS.


III. Dendrograms

IV. Spine counts distribution

Left: 3D reconstruction of CA1 pyramidal cells of mouse brain. Right: Dendrograms of basal (red) and apical (blue) dendrites of pyramidal cells. Bottom: Total lengths of apical and basal dendritic branches.

Left: Golgi-impregnated dentate gyrus & hilus (10x). Right: Representative image of spine counts (red dots) of a dendritic segment.   Bottm: Comparison of spine density (spine # / um3) between transgenic and wildtype mice 
Based on digitized serial images and quantitative data of Golgi-impregnated neurons, 3D reconstruction will provide more detailed information of neuronal properties of CNS.



V. Sholl analysis

VI. 3D dendritic morphology

Frequency of intersections of basal dendrites of CA1 pyramidal cells.

3D reconstructed dendritic trees & spines of a pyramidal cell in the mouse cortex (*Spines can be visualized by third-party softwares)



For data analysis, we prefer the subject IDs to be masked or blinded.  This will avoid potential evaluator's bias during the process of analysis.  Analysis of Variance (ANOVA) test will be performed and submitted to the Sponsor, with a CD that contained representative images (a low- and high-mag images per region).
Terms and Conditions

  • For quality assurance of our service, it is recommended that you discuss with us for preferred perfusion protocol and histology and/or immunolabeling protocols.
  • It is suggested that you use Gel-coated microscopic slides for tissue mounting and 0.17um-thick coverslips.
  • A 15% of the fee will be due upon authorization of the study; and the remaining fee will be due upon delivery of study results.
  • Progress of the service is contingent upon staining quality of tissues, operated by the independent contractor.
  • Should early termination occur, Neurodigitech will prorate the cost incurred and invoice the Sponsor. The first portion of the fee is non-refundable.