New strategies for embedding, orientation and sectioning of small brain specimens enable direct correlation to MR-images, brain atlases, or use of unbiased stereology.
We present a newly developed brain slicing machine and technique for tissue embedding, which enable orientation of fresh or fixed brain tissue from small laboratory animals, in any given position, and subsequent tissue sectioning into slabs with an optional thickness between 0.5 and 20 mm. The oriented tissue slabs may be analysed directly, or processed further on a cryostat or vibratome, into thin stainable histological sections, and aligned to MR-images or brain atlases, depending on the reference used for the initial orientation. Additionally, we describe a new embedding medium (HistOmer) which is an alginate cold polymer ready for instant use after mixing with water. HistOmer allows accurate positioning of the tissue during embedding, and at the same time supports and protects the embedded tissue during sectioning. HistOmer is, therefore, described comprehensively and compared with other commonly used embedding media. This novel slicing technique may also, as illustrated, be used to perform isotropic random orientation of the embedded tissue, before sectioning into tissue slabs of the same thickness. The technique thereby fulfills the requirements for optimal tissue sampling and preparation needed for an unbiased stereologic analysis.
Bjarkam
C R
CR
Department of Neurobiology, Institute of Anatomy, University of Aarhus, DK-8000 Aarhus C, Denmark. cb@ana.au.dk
Pedersen
M
M
Sørensen
J C
JC
D000464:Alginates / Q000737:chemistry; D000818:Animals; D048588:Benzoxazines; D001921:Brain / Q000166:cytology* / Q000378:metabolism; D002452:Cell Count / Q000295:instrumentation / Q000379:methods; D004396:Coloring Agents; D008279:Magnetic Resonance Imaging / Q000295:instrumentation / Q000379:methods; D008297:Male; D008867:Microtomy / Q000295:instrumentation / Q000379:methods*; D010078:Oxazines; D011108:Polymers / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D016610:Tissue Embedding / Q000295:instrumentation / Q000379:methods*
D000464:Alginates; D048588:Benzoxazines; D004396:Coloring Agents; D010078:Oxazines; D011108:Polymers; C028911:cresyl violet