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Non-human Primate Anatomical Research Platform
 

The Anatomical Research Platform of ION was established in 2017 for studying the mesoscopic connectome of Non-human Primate. It is located in 320 Yueyang Road, Shanghai, China.

A connectome is a comprehensive map of the connections of the nervous system. The Connectomics Platform is more specifically concerned with the investigation of the cortical connectome at the meso-scale. That is to say the comprehensive set of connections linking cortical areas in model organisms. The fact that the connectome deals with comprehensive maps, means that there is a need to develop high-throughput analysis of connectivity. The plateform is geared to analyzing the inputs to defined cortical areas that are determined using high-fidelity, high-sensitivity retrograde tracers.

The cortical connectome can be viewed as a graph and the analysis of the cortical graph exploits definitions and algorithms used in graph theory. The density of the cortical graph is very high (macaque 67%, mouse 97%). However, the range of weights of connections span 5 to 6 orders of magnitude, meaning that binary, connected/not-connected measurements are of little value. The determination of the weighted cortical graph requires accurate counts of labeled neurons in order to generate reliable and consistent connectivity profiles of individual areas.

The Connectomics Platform consists of three interrelated facilities: (i) a customized surgical suite for injection of tracers; (ii) a histology lab for preparing material for microscopic examination; (iii) microscope facilities.

Surgical suite  

Fully equipped for investigating the anesthetized macaque in a stereotaxic apparatus (respirator, CO2 monitor, blood pressure and CO2 monitor, heating blanket). Tracer injections are made using BrainSight® technology from Rogue Research Inc, a brain navigator tool that exploits MRI scans acquired in the 3-T Siemens MRI scanner.

Histology lab

Tissue is prepared in a fully equipped histology lab with chemicals and glassware. The histology lab has linear hooded mounting desks for mounting sections and freezers and refrigerators for storing antibodies, and appropriate reagents for immunohistochemistry. The histology lab is equipped with a large microtome for cutting NHP brains. The Microm HM 440 E Microtome is an ergonomically-designed sliding microtome. It employs electronic control systems and high-precision mechanics to achieve optimal section quality and reproducibility of paraffin and frozen specimens. It can accommodate specimen sizes up to 80 x 60 mm, and can cut sections ranging from 1 to 100 microns.

Microscope facilities

Our automated imagery stations exploit PlotFast® software for neuronal cartography in order to explore projection pathways. Analysis consists in the recording of the XY coordinates of very large numbers of neurons using a digitalized imaging system equiped with automatic cell counting. 2D cartography of neuronal labeling is used to reconstruct 3D (Map3D) maps to obtain accurate data on individual neuron pathways length, important for modeling networks of the cerebral cortex.
In addition to charting neurons using PlotFast® technology we also use slide scanning technology. Zeiss AxioscanZ1 (off site) uses multiple channels to create virtual digital copies of sections with up to 4 different tracers that are detected at different wave-lengths and subsequently analyzed using PlotFast
  
Data Analysis

The analysis of the structural connectivity network is done using the free software environment for statistical computing and graphics R (https://cran.r-project.org/). In particular, the majority of network properties are investigated using network analysis tools available in the R package igraph (http://igraph.org/r/). For data visualization we mainly use the packages lattice (https://cran.r-project.org/web/packages/lattice/index.html) and latticeExtra (https://cran.r-project.org/web/packages/latticeExtra/index.html). We use the package stats (https://stat.ethz.ch/R-manual/R-devel/library/stats/html/00Index.html) for additional statistical calculations. We are currently developing an R toolbox for analyzing complex structural networks (directed, undirected, weighted and unweighted), in particular to compute commonly used measures of centrality, segregation and influence, at the local and global levels.


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