Sojka M, Davies H A, Rusakov D A, Stewart M G
Department of Biology, Open University, Milton Keynes, UK.
J Neurosci Methods. 1995 Nov;62(1-2):73-82. doi: 10.1016/0165-0270(95)00056-9.
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.
