Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (34): 5492-5496.doi: 10.3969/j.issn.2095-4344.1414

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Comparison of the fixation effects of three kinds of fixatives on mouse eyeball tissue

Sun Liang1, Xu Lin2
  

  1. 1Department of Critical Care Medicine, 2Central Laboratory, Shandong Provincial Third Hospital, Jinan 250000, Shandong Province, China
  • Received:2019-05-09 Online:2019-12-08 Published:2019-12-08
  • Contact: Xu Lin, Engineer, Central Laboratory, Shandong Provincial Third Hospital, Jinan 250000, Shandong Province, China
  • About author:Sun Liang, Attending physician, Department of Critical Care Medicine, Shandong Provincial Third Hospital, Jinan 250000, Shandong Province, China

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Abstract:

BACKGROUND: Fixative is a major factor affecting the quality of pathological sections. To optimize the conditions of pathological observation of eyeball tissue, a relatively simple, practical, and reliable fixative is required.
OBJECTIVE: To compare the effects of three fixative solutions on paraffin sections of mouse eyeball tissues and optimize the fixation methods.
METHODS: Thirty eyeballs of BALB/c mice (Jinan Pengyue Experimental Animal Breeding Co., Ltd., China) were randomly divided into three groups (n = 10 eyeballs/group). The harvested eyeballs were fixed in three kinds of fixatives separately, namely formaldehyde-glutaraldehyde fixative (group A), Davidson’s fixative (group B) and 10% formaldehyde fixative (group C). Twenty-four hours later, specimens were prepared for hematoxylin-eosin staining, and the fixation effects were compared. Animal experiments were approved by the Animal Ethics Committee, Shandong Provincial Third Hospital, China (approval No. sdsldsyy-m-201801006).
RESULTS AND CONCLUSION: The eyeballs of each group were all slightly contracted and deformed. Hematoxlyin-eosin staining revealed that in the group A, the eyeball wall was shrunk; the tissue structure of the lens was relatively complete and clear without fragments in the center, and cells arranged well; large gaps appeared between corneal fibroblasts; retinal tissue was clearly detached and separated, with cells in each layer unclear; obvious blank fissures were visible between cells. In the group B, the retinal was slightly thinned, the layered arrangement was flat, with no obvious loss or shedding; the cells in each layer were arranged neatly, the nuclear membrane of each layer of the nucleus was clear, the cells did not shrink or expand; the layers of the cornea were intact and had no obvious structural changes, with no peeling phenomenon; the structure of each corneal layer was clear with no crack; different degrees of fragmentation and peeling were observed in the center of the lens, with discontinuous staining. In the group C, the corneal tissue was slightly contracted, the layers of the corneal epithelium were relatively neat, the layers were relatively clear; the lens and retinal structure were basically intact, but the lens tissue shrank and ruptured; vacant space formed, cells were arranged in disorder, retinal tissue fell off, the layers were arranged evenly; cell structure was basically clear, and obvious blank fissures were visible between cells. These results suggest that for hematoxylin-eosin staining, Davidson’s fixative is ideal for the fixation of the cornea and retina and formaldehyde-glutaraldehyde fixative is better for fixing mouse eyeball lens tissue.   

Key words: mouse eyeball, paraffin section, hematoxylin-eosin staining, formaldehyde-glutaraldehyde fixative, Davidson’s fixative, 10% formaldehyde, cornea, lens

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