The application of Whole-Mounting immnunofluorescent staining and laser speckle flow imaging technology in the study of extended mouse-ear flap model
10.3760/cma.j.cn114453-20190621-00198
- VernacularTitle:全组织包埋免疫荧光染色技术与激光散斑血流成像仪技术在小鼠耳跨区皮瓣动物模型研究中的应用
- Author:
Peisen LIN
1
;
Lei TIAN
;
Jiting XIAO
;
Qiming MA
;
Fang FANG
;
Yuehong ZHUANG
;
Yun XIE
Author Information
1. 福建医科大学第一临床医学院创伤骨科，福州 350005
- Keywords:
Immunofluorescence technique;
Mouse;
Surgical flaps;
Hemodynamics
- From:
Chinese Journal of Plastic Surgery
2020;36(7):802-809
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the characteristics and advantages of Whole-Mounting immnunofluorescent staining and laser speckle flow imaging technology in the vascular imaging of mouse ear extended flap.Methods:In this study, total of 25 ICR mice were included.Ten ICR mice were cut off the middle and lateral angiosome to establish an extended flap model, and 3 days later, the changes in the blood supply of the ear flap were observed. The ear area , tissue layer thickness and blood vessel distribution in the healthy side were observed at the same time.Obtain the mouse ears 3 days after modeling, and dissect them into three layers, i.e, the anterior skin layer , the cartilage layer and the posterior skin layer. The distribution and morphology of blood vessels, nerves and monocytes/macrophages in the anterior skin layer were stained and detected by the whole-mount immunofluorescence staining.Ten mice were adopted and an incision was made through the ear horizontally above the bifurcation of the middle angiosome of the mouse ear to establish a delayed extended flap model. Then the blood flow changes in the mouse ear were observed by laser speckle flow imaging andt he blood perfusion values were recorded immediately, 1 d, 2 d, 3 d and 4 d after the operation, respectively.Results:The area of the mouse ear was about 1.3 cm 2 , the thickness was about (0.16±0.04) mm, and the blood was supplied by three vascular bundles: the lateral caudal vascular bundle, the middle vascular bundle and the medial cephalic vascular bundle. The thickness of the anterior and posterior skin and cartilage of the mouse ears were (88±5)μm, (41±3)μm and (29±2)μm, respectively. The whole-mount immunofluorescence staining results clearly showed that the diameter of small vessels in the choke area was (50 ± 6) μm on the third day after modeling. It could be seen that the nerve and artery in mouse ear were in concomitant relationship and the nerve segment attached to the surface of the artery without obvious accompany or clinging to the vein. There were a large number of monocyte macrophages distributed in clusters in the dilated and curved arteries, but they were only scattered outside the artery. Laser speckle flow imaging results showed that there were (6 ± 2) transverse vessels in each auricular flap, and the diameter and blood flow increased significantly in the delayed extended earflap model. Immediately after the operation and at 1d, 2d, 3d, and 4d, the average blood perfusion values of transverse vessels were (92±11) PU, (136±26) PU, (147±27) PU and (176±27) PU, respectively. Conclusions:The Whole-Mounting immnunofluorescent staining and the laser speckle blood flow imaging technology can be used to well observe the blood vessels, nerves, mononuclear macrophages and blood flow perfusion of the mouse extended flap, which can play an important role in the study of blood supply of mouse extended flap.
