HEMATOXYLIN AND EOSIN (H & E) Staining

- corner stone of tissue-based diagnosis.

- plays a significant role in tissue-based diagnosis by coloring otherwise transparent tissue
sections, and allowing cell structures including the cytoplasm, nucleus, and organelles and extra-
cellular components to be clearly visible under the microscope.

HISTOCHEMICAL STAINING (HISTOCHEMISTRY)

- process whereby various constituents of tissues are studied thru chemical reactions that will
permit microscopic localization of a specific tissue substance
 EXAMPLES OF STAIN USED
 Perl's Prussian blue reaction for hemoglobin
 Periodic Acid Schiff staining for carbohydrates.

IMMUNOHISTOCHEMICAL (IHC) STAINING

- widely used in the diagnosis of abnormal cells such as those found in cancerous tumors, in the
localization of biomarkers and differentially expressed proteins in different parts of a biological
tissue, and in the detection of specific molecular markers that are characteristic of particular
cellular events such as proliferation or cell death (apoptosis)
- detect and demonstrate tissue antigens (e.g., proteins) and phenotypic markers under the
microscope.
- The current recommendation for immunohistochemical techniques is a maximum of 4% neutral
buffered formaldehyde solution
- FIXATION TIME: maximum of 48 hours

STAINS AND STAINING SOLUTIONS

- Biological stains or coloring substances are prepared from dyes which may generally be divided
into two categories:
1. Natural dyes - e.g., cochineal dyes, logwood dyes, and vegetable extracts
2. Synthetic (artificial) dyes - e.g., aniline or coal tar dyes

NATURAL DYES

- obtained from plants and animals, previously utilized for dyeing of wool and cotton. Among the
most common natural dyes available are:
1. Hematoxylin
2. Cochineal dyes and its derivatives
3. Orcein
4. Saffron

I. HEMATOXYLIN

- natural dye derived by extraction from the core or the heartwood of a Mexican tree known as
"Hematoxylin Campechianum”

- most valuable staining reagent

- not a true basic dye