Application of Novel Environmentally Friendly Reagents in Pathological Section Preparation and Immunohistochemistry
10.11714/jsysu.med.YX20250190
- VernacularTitle:新型环保试剂在病理制片及免疫组化中的应用
- Author:
Jieying LI
1
;
Shujun LIANG
1
;
Ying DING
1
;
Shuying LIU
1
;
Lina QIN
1
;
Zhaoliang PAN
1
;
Lianqiang FENG
1
;
Huijie YUAN
1
Author Information
1. Basic Medical Experimental Teaching Center, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, 510080, China
- Publication Type:Journal Article
- Keywords:
environmentally friendly reagents;
paraffin section;
HE staining;
special staining;
immunohistochemistry
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2026;47(3):509-517
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the application effects of environmentally friendly reagents versus traditional reagents in tissue processing, paraffin sectioning, hematoxylin-eosin (HE) staining, special staining, and immunohistochemistry. MethodsPancreatic, renal, splenic, hepatic, pulmonary, skeletal muscle, and cardiac tissues were collected from mice and New Zealand rabbits, and randomly divided into a traditional group (Group A), an environmentally friendly reagent I group (Group B), and an environmentally friendly reagent Ⅱ group (Group C). The processed tissue sections from all three groups were subjected to paraffin sections, HE staining, phosphotungstic acid-hematoxylin staining, and Mallory trichrome staining. The pancreatic tissue sections were subjected to immunohistochemical detection of insulin protein. Evaluation criteria included tissue morphological integrity, embedding operability, section quality, and staining effects. ResultsNo significant differences were observed among the three groups of samples in terms of fixation efficacy, embedding operations, and section quality. After HE staining, all groups exhibited bright nuclear and cytoplasmic staining with clear contrast, with excellent-and-good rates of 98%, 99%, and 97% for Groups A, B, and C, respectively, showing no statistically significant differences (A vs. B, P=0.561; A vs. C, P=0.651). In phosphotungstic acid-hematoxylin staining, Groups A and C showed clear structural contrast and bright staining for striated muscle and collagen fibers, whereas Group B exhibited uneven staining. The excellent-and-good rates were 100%, 10%, and 95% for Groups A, B, and C, respectively, with significant differences between Group A and Group B (P<0.01) but no statistical difference between Group A and Group C (P=0.311). In Mallory trichrome staining, Groups A and B outperformed Group C. The excellent-and-good rates were 95%, 90%, and 15% for Groups A, B, and C, respectively, with significant differences between Group A and Group C (P<0.01), but no statistical difference between Group A and Group B (P=0.548). Immunohistochemical results showed that Group A had a clean background with no non-specific staining, while Groups B and C exhibited significant non-specific staining, with statistically significant differences in non-specific staining rates between the groups (A vs. B, 5% vs. 100%, P<0.01; A vs. C, 5% vs.100%, P<0.01). ConclusionIn HE staining, environmentally friendly reagents demonstrated effective tissue fixation and staining results comparable to traditional reagents, meeting the teaching requirements. For special staining, Group B and Group C performed better in Mallory trichrome staining and phosphotungstic acid-hematoxylin staining, respectively. However, environmentally friendly reagents exhibited non-specific staining issues in immunohistochemistry, where further optimization is required.