An optimized method for embedding undecalcified mouse tibias in plastic blocks.
10.12122/j.issn.1673-4254.2019.09.06
- Author:
Zhonghao DENG
1
;
Jingde LIN
2
;
Zheting LIAO
1
;
Yufan CHEN
1
;
Desheng WU
1
;
Shuhao FENG
1
;
Nachun CHEN
1
;
Baohong ZHAO
3
;
Liang ZHAO
1
Author Information
1. Department of Orthopedic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
2. Department of Orthopaedics, Anxi Traditional Chinese Medicine Hospital, Quanzhou 362400, China.
3. Arthritis and Tissue Degeneration Program and The David Z. Rosensweig Genomics Research Center, NewYork 10021, USA.
- Publication Type:Journal Article
- Keywords:
multiple tibias embedding;
plastic embedding;
slide detachment;
undecalcified bones
- MeSH:
Animals;
Mice;
Plastics;
Staining and Labeling;
Tibia;
Tissue Embedding;
methods
- From:
Journal of Southern Medical University
2019;39(9):1038-1044
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To optimize the method for embedding multiple undecalcified mouse tibias in plastic blocks, improve the efficiency and stability of plastic embedding and reduce the detachment rate of plastic slides.
METHODS:Thirty undecalcified tibias from 15 B6 mice were used for plastic embedding after calcein labeling, fixation, dehydration and infiltration. The tibias were embedded in cylindrical plastic blocks with a diameter of 4 mm. For each bone, the 1/4 proximal tibia was cut off, and the remaining 3/4 was used for re-embedding. Five bones were embedded in a single block with each bone standing closely on the surface of a flat plate. The samples were randomized into control and experimental groups in all the processes of embedding, sectioning and staining. In the 3 groups with modified embedment, flowing CO was added into the embedding solution, embedding solution was applied to the section surface, and the slides were heated at 95 ℃ for 15 min. The polymerization time, slide detachment rate, bone formation and osteoblast parameters were analyzed.
RESULTS:We prepared 6 plastic blocks, each containing 5 tibias, whose cross sections were on the same plane. The blocks were completely polymerized and suitable for sectioning. Flowing CO into the embedding solution reduced the polymerization time and increased the rate of complete polymerization. Application of the embedding solution on the section surface significantly reduced the detachment rate of the sections ( < 0.05) without affecting bone formation analysis ( > 0.05). Heating the slides significantly lowered the detachment rate of the sections ( < 0.05) without affecting osteoblast analysis ( > 0.05).
CONCLUSIONS:The optimized method allows effective embedding of multiple undecalcified mice tibias in the same block and can be an ideal method for histological analysis of undecalcified bones.
