Comparative study on the fixation effect of different tissue fixatives on proteins in hair follicles
Objective To investigate the effects of 4%paraformaldehyde,10%neutral buffered formaldehyde solution,and 95%ethanol as four types of tissue fixatives on immunofluorescence staining of frozen sections of rat hair follicles,and identify the optimal fixative for frozen sections.Methods Two SD rats were randomly selected and anesthetized with ether.The skin of the rat lips was cut off,and the tentacle hair follicles were isolated and immediately performed serial frozen sections.The sections were fixed in four different fixatives including 4%paraformaldehyde,10%neutral buffered formaldehyde solution,acetone and 95%ethanol.After the same fixation time,all sections were subjected to immunofluorescence staining to detect the expression of CK15 and β-catenin proteins in the hair follicles.Results The immunofluorescence staining results showed that the preservation effect of four different fixatives on proteins were different.Among them,4%paraformaldehyde had better preservation effect,and strong fluorescence expression was observed in the tissues.However,tissues fixed with 10%neutral buffered formaldehyde solution,acetone and 95%ethanol showed lower expression levels.Conclusion There are differences in the preservation effect of different tissue fixatives on proteins,and 4%paraformaldehyde has better preservation effect on proteins in hair follicles.
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