Embedding of large specimens in glycol methacrylate: prerequisites for multi-signal detection and high-resolution imaging.
Acrylic resin mixtures are commonly used to study microscopic sections of biological specimens, giving the advantage of good morphological preservation. Existing embedding protocols, however, are suitable for tissue blocks, not exceeding 1 mm in thickness. We have developed a protocol to embed larger specimens (up to 2 cm(3)) in Technovit 8100. This medium allowed us to perform classic histological (trichrome), silver, as well as immunohistochemical staining, needed for multi-signal detection at high-resolution imaging to reconstruct a three-dimensional interpretation of a serially sectioned muscle. The technique was applied to reconstruct the semitendinosus muscle of a fetal pig, 44 days post conception, featuring connective tissue, intramuscular nerves, blood vessels, and muscle fibre types. For the reconstruction, a technique was used that enabled us to insert high-resolution images of histological details into low-resolution images of the entire muscle.
De Jonge
Henriëtte W
HW
Faculty of Veterinary Medicine, Department of Pathobiology, Anatomy Division, PO Box 80.158, 3508 TD, Utrecht, The Netherlands. h.w.dejonge@vet.uu.nl
De Bakker
Merijn A G
MA
Verbeek
Fons J
FJ
Weijs
Wim A
WA
D000818:Animals; D005865:Gestational Age; D006614:Hindlimb; D007089:Image Enhancement; D021621:Imaging, Three-Dimensional; D007150:Immunohistochemistry; D008689:Methacrylates; D018482:Muscle, Skeletal / Q000033:anatomy & histology* / Q000196:embryology; D016613:Plastic Embedding / Q000379:methods*; D013552:Swine; D013979:Tibial Nerve / Q000033:anatomy & histology
D008689:Methacrylates; C005044:hydroxyethyl methacrylate