3-dimensional morphometry of intact dendritic spines observed in thick sections using an electron microscope.
An experimental technique is described which allows observation of fixed neuronal dendrites at magnifications from 10-12 K. The method uses 4-7-microns-thick sections of Epon-embedded tissue with nerve cells that are first impregnated by the rapid Golgi technique and then stained with gold particles/aggregates using a modified gold-toning procedure. A relatively high acceleration voltage (200 kV) is employed to observe in fine detail the dendritic fragments of interest at different angular positions in space, by using a eucentric goniometer stage with a tilt angle of +/- 45 degrees. Image analysis methodology is proposed which permits estimation of 3-dimensional (3D) lengths and of the volume of observed intact dendritic spines. The advantages of the technique with respect to 3D reconstruction methodology are discussed.
Sojka
M
M
Department of Biology, Open University, Milton Keynes, UK.
Davies
H A
HA
Rusakov
D A
DA
Stewart
M G
MG
D000818:Animals; D048429:Cell Size / Q000502:physiology; D002645:Chickens; D003712:Dendrites / Q000648:ultrastructure*; D008433:Mathematics; D008854:Microscopy, Electron / Q000379:methods*; D008867:Microtomy; D013194:Staining and Labeling