Hematoxylin-eosin stain

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HE staining of pancreatic tissue with islet of Langerhans (light area)
Histological section of the endometrial glands after HE staining

The hematoxylin-eosin staining, often abbreviated as HE staining , is a staining method in histology with which the various structures of a tissue section can be stained.

properties

The hematoxylin - eosin staining is used to differentiate between different tissue structures in the microscopic image using two different individual stains and is one of the most widely used routine staining methods for morphological examinations . In pathology , pathological changes in biopsies and surgical specimens can be examined with the help of this overview color . The method takes between five and 45 minutes, depending on the protocols and staining solutions used, and is often part of extensive tissue processing that takes one to several days. HE staining is also used in various ways in research, in particular as an overview staining before making immunohistochemical stains to examine various aspects, such as B. in the diagnosis of tumors . Tumors with a diameter of less than 0.2 mm can sometimes be overlooked, which is why hematoxylin-eosin staining is often used in parallel with immunostaining .

Hematoxylin is a natural dye from the bluewood tree . In order to develop its coloring properties, it must be oxidized to hematein. Alaunhämatoxylin, a basic Hämateinlack as "hemalum" in the histological technique often used, colors all acidic or basophilic structures blue, particularly cell nuclei with the contained deoxyribonucleic acid (DNA) and with ribosomes enriched rough endoplasmic reticulum (RER, English rough endoplasmic reticulum ).

Eosin (more precisely Eosin Y) is a synthetic acidic dye and stains all acidophilic or basic (eosinophilic) structures red, especially cell plasma proteins , mitochondria , the smooth endoplasmic reticulum (sER, English smooth endoplasmic reticulum ), collagen and keratin .

After the hematoxylin staining, the cell nuclei initially appear reddish-brown due to the low pH of the staining solution. When the pH value is increased (blueing), the color changes to the typical blue-violet; this is often done simply by rinsing in tap water , but there are also special buffer solutions for this purpose ( Scott buffers ), since waterworks reserve the right to chlorinate the water if necessary, and the chlorine destroys the color. This is followed by staining the cytoplasm in an alcoholic or aqueous solution of eosin. The water is displaced from the tissue section by further rinsing steps using alcohol solutions in increasing concentrations up to absolute alcohol. Finally, the drained section is cleared in an organic solvent such as xylene and can now be covered with a mounting medium and a cover slip . In this way, the cut and color are preserved for decades and can be microscoped.

The hematoxylin-eosin stain can be combined with an auramine O stain. With some restrictions, DNA can be extracted from a colored specimen .

literature

  • Godwin Avwioro (2011): Histochemical Uses Of Haematoxylin - A Review. In: International Journal of Research and Reviews in Applied Sciences. Vol. 1, pp. 24-34. PDF

Web links

Individual evidence

  1. ↑ JM Haggerty, XN Wang, A. Dickinson, CJ O'Malley, EB Martin: Segmentation of epidermal tissue with histopathological damage in images of haematoxylin and eosin stained human skin. In: BMC medical imaging. Volume 14, 2014, p. 7, doi : 10.1186 / 1471-2342-14-7 . PMID 24521154 . PMC 3942169 (free full text).
  2. ↑ SM Giobuin, DO Kavanagh, E. Myers, AO Doherty, CM Quinn, T. Crotty, D. Evoy, E. McDermott: The significance of immunohistochemistry positivity in sentinel nodes which are negative on haematoxylin and eosin in breast cancer. In: European Journal of Surgical Oncology . Volume 35, Number 12, December 2009, pp. 1257-1260, doi : 10.1016 / j.ejso.2009.04.004 . PMID 19497702 .
  3. ↑ S. Ramulu, AD Kale, S. Hallikerimath, V. Kotrashetti: Comparing modified papanicolaou stain with ayoub-shklar and haematoxylin-eosin stain for demonstration of keratin in paraffin embedded tissue sections. In: Journal of oral and maxillofacial pathology: JOMFP. Volume 17, number 1, January 2013, pp. 23-30, doi : 10.4103 / 0973-029X.110698 . PMID 23798825 . PMC 3687183 (free full text).
  4. ↑ P. Trenchev, M. Konopka: Combined haematoxylin, eosin and fluorescent stain for paraffin sections. In: Pathology. Volume 12, Number 1, January 1980, pp. 79-81, PMID 6154918 .
  5. ↑ DP Jackson, S. Bell, J. Payne, FA Lewis, J. Sutton, GR Taylor, P. Quirke: Extraction and amplification of DNA from archival haematoxylin and eosin sections and cervical cytology Papanicolaou smears. In: Nucleic acids research. Volume 17, Number 23, December 1989, p. 10134, PMID 2481259 . PMC 335270 (free full text).