Chemical Preservation

Brief overview on the state of the art in chemical preservation research:
ProposalForBrainPreservationTechnologyPrize.pdf

Papers describing the state of the art in chemical brain preservation:

Palay et al. 1962. Fixation of neural tissues for electron microscopy by perfusion with solutions of osmium tetroxide.
http://jcb.rupress.org/cgi/reprint/12/2/385

Oldmixon et al. 1985. Perfusion dehydration fixes elastin and preserves lung air-space dimensions.
http://jap.physiology.org/cgi/reprint/58/1/105

Excellent reference books describing theory and protocols for perfusion fixation of tissue and solid organs:

Hyat, M.A. 2000. Principles and Techniques of Electron Microscopy: Biological Applications, Fourth Ed., Cambridge U. Press.
Kiernan, J.A. 2008. Histological and Histochemical Methods: Theory and Practice, Fourth Ed., Cold Spring Harbor Lab Press.

Cryopreservation

Brief overview on the state of the art in cryopreservation research:
ProposalForBrainPreservationTechnologyPrize.pdf

Papers describing the state of the art in cryopreservation:

Darwin et al. 1995. Effect of human cryopreservation protocol on the ultrastructure of the canine brain.
http://www.alcor.org/Library/html/braincryopreservation2.html

Pichugin et al. 2006. Cryopreservation of rat hippocampal slices by vitrification.
http://www.ncbi.nlm.nih.gov/pubmed/16403489?dopt=Abstract

Fahy et al. 2009. Physical and biological aspects of renal vitrification.
http://www.21cm.com/abstracts.jsp

Connectomics, Imaging, and Neural Circuit Mapping

There are a number of imaging and neural circuit mapping technologies that are rapidly improving in price/performance today. Any one of these might be used to complete a small animal (worm, insect, zebrafish), a mouse, and later, a human connectome in coming years. Which will ultimately be proven most cost effective is not clear at present, and may vary from application to application and lab to lab. Current viable technologies include:

1. Serial Block Face Scanning Electron Microscopy (SBFSEM)
2. Focused Ion Beam Scanning Electron Microscopy (FIBSEM)
3. Ultrathin sectioning and automated tape collection for SEM
4. Electron tomography on thick cut sections (TEM or SEM)
5. X-ray tomography (either on vitrified tissue or stained plastic embedded tissue)
6. Atomic force scanning microscopy of block face in preserved tissue.

BPF Brief on The NIH Human Connectome Project:
What is a connectome?

Brief overview on the state of the art in high-throughput volume EM imaging of brain tissue:
ProposalForBrainPreservationTechnologyPrize.pdf

Papers describing the state of the art in high-throughput volume EM imaging of brain tissue

Denk et al. 2004. Serial block-face scanning electron microscopy to reconstruct three-dimensional tissue nanostructure.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC524270/

Knott et al. 2008. Serial section scanning electron microscopy of adult brain tissue using focused ion beam milling.
http://www.jneurosci.org/cgi/content/full/28/12/2959

ATLUM project website:
http://www.mcb.harvard.edu/lichtman/ATLUM/ATLUM_web.htm

Papers describing recent imaging technology advances:
Schneider et al. 2010. Three-dimensional cellular ultrastructure resolved by X-ray microscopy.
http://www.nature.com/nmeth/journal/v7/n12/full/nmeth.1533.html