梨石蜡切片制备方法的优化

doi:10.16420/j.issn.0513-353x.2024-0849

摘要/Abstract

摘要： 在常规石蜡切片制备方法基础上，以梨叶片、新梢和花芽等为试材，对脱水、透明、粘片和染色等步骤进行了优化。结果表明：采用微波水浴方法处理，制片时间（脱水、透明和浸蜡）由原750 ~ 1 320 min缩短至48 ~ 108 min；对3种透明剂进行筛选，浸蜡脱蜡透明液透明效果较好，可减少二甲苯使用；采用水浴粘片方法，将展片粘片合为一步，可简化操作步骤；4种染色方法均能很好的染色，可根据试验目的选择适宜的方法。最终形成一套微波水浴石蜡切片方法，该方法可缩短制备时间，提高制片效率。

关键词: 梨, 石蜡切片, 微波水浴

Abstract:

Based on the conventional paraffin section method，using pear leaves，shoots，and flower buds as experimental materials，optimized key steps—including dehydration，clearing，pasting and staining. The results demonstrated that employing microwave and water bath processing dramatically reduced the total processing time（encompassing dehydration，clearing and dipping paraffin）from 750-1 320 min to 48-108 min. Screening of three clearing agent revealed that the clearing agent used for dipping paraffin and dewaxing solution demonstrated superior clearing efficacy，while significantly reducing xylene usage. The water bath mounting method combined tissue flattening and adhesion into a single step，simplifying the procedure. Comparative evaluation of four staining methods allowed selection of the most suitable protocol based on specific sectioning requirements. Ultimately，developed an integrated microwave and water bath paraffin sectioning method. This method significantly shortens preparation time and enhances sectioning efficiency.

Key words: pear, paraffin section method, microwave and water bath method

武春昊, 闫兴凯, 卢明艳, 赵滢, 张茂君, 李思宇, 王强. 梨石蜡切片制备方法的优化[J]. 园艺学报, 2025, 52(9): 2554-2564.

WU Chunhao, YAN Xingkai, LU Mingyan, ZHAO Ying, ZHANG Maojun, LI Siyu, WANG Qiang. Optimization of Paraffin Section Preparation Method for Pear Tissues[J]. Acta Horticulturae Sinica, 2025, 52(9): 2554-2564.

图/表 11

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步骤 Step	试剂梯度 Reagent gradient	叶片 Leaf		新梢 Shoot		花芽 Flower bud
步骤 Step	试剂梯度 Reagent gradient	CPSM	MWBM	CPSM	MWBM	CPSM	MWBM
脱水 Dehydration	70%乙醇 70% Ethanol	60	1	120	6	120	6
	85%乙醇 85% Ethanol	60	1	120	6	120	6
	95%乙醇 95% Ethanol	60	1	120	6	120	6
	100%乙醇 100% Ethanol	60	1	120	6	120	6
	100%乙醇 100% Ethanol	60	1	120	6	120	6
透明 Clearing	1/2乙醇 + 1/2二甲苯 1/2 Ethanol + 1/2 Xylene	60	1	120	6	120	6
	二甲苯 Xylene	60	1	120	6	120	6
	二甲苯 Xylene	60	1	120	6	120	6
浸蜡 Dipping paraffin	1/2二甲苯+石蜡 1/2 Xylene + Paraffin	90	10	120	15	120	15
	石蜡 Paraffin	90	10	120	15	120	15
	石蜡 Paraffin	90	10	120	15	120	15
	石蜡 Paraffin	—	10	—	15	—	15
总时长 Total time		750	48	1 320	108	1 320	108

步骤 Step	试剂梯度 Reagent gradient	叶片 Leaf		新梢 Shoot		花芽 Flower bud
步骤 Step	试剂梯度 Reagent gradient	CPSM	MWBM	CPSM	MWBM	CPSM	MWBM
脱水 Dehydration	70%乙醇 70% Ethanol	60	1	120	6	120	6
	85%乙醇 85% Ethanol	60	1	120	6	120	6
	95%乙醇 95% Ethanol	60	1	120	6	120	6
	100%乙醇 100% Ethanol	60	1	120	6	120	6
	100%乙醇 100% Ethanol	60	1	120	6	120	6
透明 Clearing	1/2乙醇 + 1/2二甲苯 1/2 Ethanol + 1/2 Xylene	60	1	120	6	120	6
	二甲苯 Xylene	60	1	120	6	120	6
	二甲苯 Xylene	60	1	120	6	120	6
浸蜡 Dipping paraffin	1/2二甲苯+石蜡 1/2 Xylene + Paraffin	90	10	120	15	120	15
	石蜡 Paraffin	90	10	120	15	120	15
	石蜡 Paraffin	90	10	120	15	120	15
	石蜡 Paraffin	—	10	—	15	—	15
总时长 Total time		750	48	1 320	108	1 320	108

组织 Tissue	方法 Method	蜡带类型The wax bands types			蜡带成带比率/% The ratio of wax band types
组织 Tissue	方法 Method	成带（片） Success of wax bands （piece）	裂带（片） Split of wax bands （piece）	不成带（片） Fail of wax bands （piece）	蜡带成带比率/% The ratio of wax band types
叶片Leaf	CPSM	30	0	0	100.0
叶片Leaf	MWBM	30	0	0	100.0
新梢Shoot	CPSM	24	6	0	80.0
新梢Shoot	MWBM	25	5	0	83.3
花芽Flower bud	CPSM	13	14	3	43.3
花芽Flower bud	MWBM	15	10	5	50.0

组织 Tissue	方法 Method	蜡带类型The wax bands types			蜡带成带比率/% The ratio of wax band types
组织 Tissue	方法 Method	成带（片） Success of wax bands （piece）	裂带（片） Split of wax bands （piece）	不成带（片） Fail of wax bands （piece）	蜡带成带比率/% The ratio of wax band types
叶片Leaf	CPSM	30	0	0	100.0
叶片Leaf	MWBM	30	0	0	100.0
新梢Shoot	CPSM	24	6	0	80.0
新梢Shoot	MWBM	25	5	0	83.3
花芽Flower bud	CPSM	13	14	3	43.3
花芽Flower bud	MWBM	15	10	5	50.0

方法 Method	叶片 Leaf			新梢 Shoot			花芽 Flower bud
方法 Method	总片数 Total	粘片 Section adhesion	脱片 Slide detachment	总片数 Total	粘片 Section adhesion	脱片 Section detachment	总片数 Total	粘片 Section adhesion	脱片 Section detachment
水浴粘片 Water bath pasting	100	100	0	100	100	0	100	100	0
明胶粘片剂 Gelatinglycerin adhesive	100	100	0	100	90	10	100	88	12