一、实验器材及试剂

1、实验器材

2、主要试剂及货号

二、实验步骤

1、石蜡切片脱蜡至水：依次于环保脱蜡剂(1)、环保脱蜡剂(2)、环保脱蜡剂(3)中分别脱蜡10分钟，然后经无水乙醇、95%乙醇、85%乙醇、75%乙醇各5分钟。自来水冲洗1分钟。

2、苏木素染色液（Harris）染色4分钟，自来水洗2分钟，至切片上无多余染液脱出。

3、用0.8%盐酸酒精分化2秒，自来水冲洗，亦可用碳酸锂水溶液返蓝，然后水洗2分钟。

4、入伊红染液（醇溶性）染20秒，不需水洗，直接入95%乙醇调色5秒，入无水乙醇(1)、无水乙醇(2)脱水2分钟。

5、环保透明剂透明、封固、镜检。

二、结果判读

细胞核呈蓝紫色，细胞质、间质、各种纤维类呈不同程度的红色。

三、注意事项

1、石蜡切片应充分脱蜡。

2、注意细胞核的分化程度，如果着色太深，可以再次分化。

3、注意苏木素和伊红的使用程度，及时更换染液。

Experimental Report on H&E Staining

1.Instruments and key reagents

1.1Instruments

1.2Key reagents

2.Procedures

2.1Deparaffinization and rehydration.

2.1.1Tissue sections were immersed in clearer for 10min. Repeat this step two times, gently shaking off excess liquid between each step.

2.1.2Tissue sections were immersed in progressively more dilute ethanol solutions and ultimately immersed in distilled water to rehydrate the tissue: Absolute ethanol for 5min, 95% ethanol for 5min, 85% ethanol for 5min, 75% ethanol for 5min. Rinsing with distilled water for 1min.

2.2Sections were stained with hematoxylin solution (Harris) for 4 minutes and then washed with tap water for 2 minutes until no excess dye comes out of the sections.

2.3Sections were differentiated with 0.8% hydrochloric acid alcohol for 2 seconds and washed with tap water. Sections can also be treated with lithium carbonate solution to be bluer and washed with tap water for 2 minutes.

2.4Sections were stained with eosin solution (alcohol soluble) for 20 seconds without water washing. Then, sections were treated with 95% ethanol for 5 minutes and dehydrated with absolute ethanol I and II for 2 minutes.

2.5The tissue sections were transparent with xylene and then mounted with neutral balsam. Examination with microscope.

3.Results

The nucleus is blue and the cytoplasm is pink to red.

4.Attentions

4.1It is important that paraffin sections need to be fully dewaxed.

4.2If nuclei are too dark, it can be differentiated again.

5.Technical support

The technical support of H&E staining experiment is provided by Hubei BIOSSCI Biotech Co., Ltd (Wuhan Changyan Pathology technology Co., Ltd).。

Tel: 400 118 0100

Fax: +86-027-87382710

E-mail: support@biossci.com

Website: www.biossci.com