The Golgi Stain Service

III. 3D modeling of the ​quantitative neuron:

3D volumetric analysis of mouse hippocampal subfields  

Neurodigitech, a Neuro-CRO biotech, neurohistology, immunohistochemistry and Golgi-Cox Stain Service

*It is noted that our Golgi Stain Service includes 1) In-house Golgi-Cox impregnation (using FD Rapid GolgiStain Kit) and 2) Rapid Golgi impregnation protocols for various types of tissue, such as unfixed, frozen, and paraffin embedded samples.  Please consult with our team for evaluation of your protocol and study design.


Golgi Stain Service Instructions:

  • The Golgi Stain procedure is mainly performed for "un-fixed" or "fresh" blocks or brains, though our staining kit has also been used for "fixed" tissue.
  • The pre-made Golgi staining solution will be shipped to your site in advance if the study is performed at your laboratory.  After harvesting of fresh blocks/brains, a 1 to 2 wk period of the immersion/incubation of the tissue in the dark is required.  Then, the box(es) of the samples will be shipped to our site for subsequent procedures till completion of the study.
  • It is recommended to have at 5-7 brains per group x 6-7 cells per brain for analysis.  The impregnated cells per brain will be randomly selected along the a-p axis of interested ROI by our Golgi Service specialists for quantitative analysis.  


*We recommend you send us a brief description of your study (see blow), so we can help you generate a reasonable and economical quote for your study! Your email should include:




















Deliverables: 


  • The Excel spreadsheets of the raw and clustered data will be organized and submitted to the Sponsor, with the representative images (a low- and high-mag images per region). 
  • The turnaround/completion of the study will be 2-3 months, but it also depends upon the quantity of the samples.  *The minimum of the sample is n=5 per group.
  • 3D reconstruction of digitized neurons will be quoted separated from the Golgi Stain Service package.



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.
  • The 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.

IV. Volumetric measurement and estimation of cell population along the anterior-to-posterior axis of the hippocampus:

3D distribution of labeled cells in mouse dentate gyrus

Services 


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

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

With computer-generated 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 3-dimensional (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.  


I. Quantitative neuron morphology, including dendritic analysis and spine counts/density:

II. Quantitative neuron morphology, including dendritic analysis and spine counts/density: