Journal of Nippon Medical School: Vol.67 No.4 page.280

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ArticleTitle Triple-Staining to Identify Apoptosis of Hepatic Cells In Situ

AuthorList Jing Zhang¹, Hong You², Tailing Wang¹, Baoen Wang², Jidong Jia², Hironori Katayama³, Shotaro Maeda³, Ruojaio Wang⁴, Goro Asano⁴, Toshiyuki Ishiwata⁴, Zenya Naito⁴ and Munehiro Yokoyama⁴

Affiliation ¹China-Japan Friendship Hospital, ²Beijing Friendship Hospital, Beijing, P. R. China ³Division of Pathology. Tama-Nagayama Hospital, Nippon Medical School Japan ⁴Department of Pathology, Nippon Medical School, Japan

Language EN

Volume 67

Issue 4

Year 2000

Page 280-283

Received January 24, 2000

Accepted April 21, 2000

Keywords apoptosis, TUNEL, hepatocyte, hepatic stellate cell, Kupffer cell

Abstract To identify apoptosis of nonparenchymal cells in fibrotic livers, we established a triple staining method which combined immunohistochemistry for cell markers and Masson staining for collagen as well as terminal deoxynucleotidyl transferase UTP nick end labeling (TUNEL). Five μm formalin fixed, paraffin-embedded liver sections were prepared for staining. Firstly, TUNEL staining was carried out to detect apoptosis of liver cells. Then, the sections were subjected to immunohistochemistry for a-smooth muscle actin (α-SMA) or KP-1 to identify hepatic stellate cells or Kupffer cells. Finally, Masson staining was performed to show the relationship between apoptosis and collagens. In addition, we optimized different conditions of fixation, digestion and color development which may affect the results.

Correspondence to Jing Zhang, Division of Pathology, China-Japan Friendship Hospital, Beijing, P. R. China, 100029. Phone: 0086-10-64221122 ext 4213

ArticleTitle	Triple-Staining to Identify Apoptosis of Hepatic Cells In Situ
AuthorList	Jing Zhang¹, Hong You², Tailing Wang¹, Baoen Wang², Jidong Jia², Hironori Katayama³, Shotaro Maeda³, Ruojaio Wang⁴, Goro Asano⁴, Toshiyuki Ishiwata⁴, Zenya Naito⁴ and Munehiro Yokoyama⁴
Affiliation	¹China-Japan Friendship Hospital, ²Beijing Friendship Hospital, Beijing, P. R. China ³Division of Pathology. Tama-Nagayama Hospital, Nippon Medical School Japan ⁴Department of Pathology, Nippon Medical School, Japan
Language	EN
Volume	67
Issue	4
Year	2000
Page	280-283
Received	January 24, 2000
Accepted	April 21, 2000
Keywords	apoptosis, TUNEL, hepatocyte, hepatic stellate cell, Kupffer cell
Abstract	To identify apoptosis of nonparenchymal cells in fibrotic livers, we established a triple staining method which combined immunohistochemistry for cell markers and Masson staining for collagen as well as terminal deoxynucleotidyl transferase UTP nick end labeling (TUNEL). Five μm formalin fixed, paraffin-embedded liver sections were prepared for staining. Firstly, TUNEL staining was carried out to detect apoptosis of liver cells. Then, the sections were subjected to immunohistochemistry for a-smooth muscle actin (α-SMA) or KP-1 to identify hepatic stellate cells or Kupffer cells. Finally, Masson staining was performed to show the relationship between apoptosis and collagens. In addition, we optimized different conditions of fixation, digestion and color development which may affect the results.
Correspondence to	Jing Zhang, Division of Pathology, China-Japan Friendship Hospital, Beijing, P. R. China, 100029. Phone: 0086-10-64221122 ext 4213