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The Optimization of H&E and LFB Staining Technique of Frozen Sections and Its Application in Autoimmune Diseases
Qin Chaoyan1, Wang Tingting1, Shi Changjie1, Du Changsheng1,2*
1Shanghai Key Laboratory of Signaling and Disease Research, School of Life Sciences and Technology,Tongji University, Shanghai 200092, China;
2State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica,Chinese Academy of Sciences, Shanghai 201203, China
2State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica,Chinese Academy of Sciences, Shanghai 201203, China
Abstract: This study was designed to investigate the optimization of hematoxylin-eosin (H&E) staining and luxol fast blue (LFB) staining in the frozen sections and application of histopathological analysis in experimental autoimmune encephalomyelitis (EAE). Five spinal cord samples were prepared from normal C57BL/6 mice and EAE mice， and each of the sample was divided into two halves for frozen embedding and paraffin embedding relatively.In the optimized conditions， results of the H&E staining and LFB staining between frozen sections and paraffin sections were compared. In paraffin sections， a fine morphology and structure of the cells were observed in H&E and LFB staining. In frozen sections， the clarity of the cells structure which was stained by H&E and LFB was almost identical with paraffin sections. The methods of H&E and LFB staining based on frozen sections were further applied to analyze the pathological changes in EAE model， and found it could repeat the known phenomena that β-arrestin2 knockout reduced EAE pathogenesis. After optimizing， staining methods based on frozen sections can achieve a similar fine performance to paraffin sections. In addition， the procedure of frozen sections production was simple and fast.Furthermore， the staining was more time-saving efficient than paraffin sections. The optimizing methods were also could be preferably applied to analyze pathological changes of autoimmune disease， which would have a wide range of applications in the study of autoimmune diseases， especially in an animal model of multiple sclerosis.
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