A modified intracellular labelling technique for high-resolution staining of neuron in 500 microm-thickness brain slice.
- Author:
Ming-liang ZHAO
1
;
Guo-long LIU
;
Jian-feng SUI
;
Huai-zhen RUAN
;
Ying XIONG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Image Processing, Computer-Assisted; Neurons; cytology; physiology; Patch-Clamp Techniques; Rats; Rats, Sprague-Dawley; Software; Staining and Labeling; methods
- From: Chinese Journal of Applied Physiology 2007;23(2):254-256
- CountryChina
- Language:Chinese
-
Abstract:
AIMTo develop simple but reliable intracellular labelling method for high-resolution visualization of the fine structure of single neurons in brain slice with thickness of 500 microm.
METHODSBiocytin was introduced into neurons in 500 microm-thickness brain slices while blind whole cell recording. Following processed for histochemistry using the avidin-biotin-complex method, stained slices were mounted in glycerol on special glass slides. Labelled cells were digital photomicrographed every 30 microm and reconstructed with Adobe Photoshop software.
RESULTSAfter histochemistry, limited background staining was produced. The resolution was so high that fine structure, including branching, termination of individual axons and even spines of neurons could be identified in exquisite detail with optic microscope. With the help of software, the neurons of interest could be reconstructed from a stack of photomicrographs.
CONCLUSIONThe modified method provides an easy and reliable approach to revealing the detailed morphological properties of single neurons in 500 microm-thickness brain slice. Without requisition of special equipment, it is suited to be broadly applied.
