Wright's stain is a histological stain that facilitates the differentiation of blood cell types.

It is used primarily to stain peripheral blood smears and bone marrow aspirates which are
examined under a light microscope. In cytogenetics it is used to stain chromosomes to
facilitate diagnosis of syndromes and diseases. The traditional Wright’s stain, an alcoholic
solution of methylene blue and eosin Y (it has a very slightly yellowish cast. Eosin Y is a
tetrabromo derivate of fluorescein.)

1. Place 1.0 ml of the Wright Stain Solution upon the smear 1 – 3 minutes.
2. Add 2.0 ml distilled water or Phosphate buffer pH 6.5 and let stand twice as long
as in step 1.
3. Rinse stained smear with water or the Phosphate buffer pH 6.5 until the edges
show faintly pinkish-red.

There are related stains known as the buffered Wright stain, the Wright-Giemsa stain, and
the buffered Wright-Giemsa stain, and specific instructions depend on the solutions being
used, which may include Eosin Y, Azure B, and Methylene Blue (some commercial
preparations combine solutions to simplify staining). The May-Grünwald stain, which
produces a more intense coloration, also takes a longer time to perform.
La tinción de Wright es una tinción de tipo Romanowsky. Una tinción de Romanowsky
consiste en azul de metileno y sus productos de oxidación, así como eosina Y o eosina B

Wright's stain is a neutral stain produced by the interaction of an acidic and a basic dye,
producing a large salt molecule with a colored dye in both its parts. The Romanovsky-type
mixtures, (including Wright's and Giemsa stains) are the best known of these neutral stains
and they are formed by the interaction of methylene blue and eosin.

With the Wright's stain, blood cells exhibit four major staining properties that allow the cell
types to be distinguished. Basophilia (affinity for methylene blue), azurophilia (affinity for
the oxidation products of methylene blue called azures, which are reddish purple),
acidophilia (affinity for eosin), and neutrophilia (affinity for a complex of dyes in the
mixture, which are pale lilac). In a stained blood smear, erythrocytes bind eosin and appear
orange to pink, nuclei purplish blue, basophilic granules very dark bluish purple,
eosinophilic granules red to red-orange, neutrophilic granules reddish-brown to lilac,
platelets violet to purple, and lymphocyte cytoplasm stains pale blue.
OJO
En conclusión, parece que la reactividad del aldehído-fucsina y el reactivo de Schiff se parecen en
que ambos muestran una afinidad por los grupos aldehído, pero diferente, ya que aldehído-fucsina
parece poseer una fuerte afinidad por los ácidos de azufre, que es el reactivo de Schiff no.

Expected results

 Elastic fibres  –  purple
 Mast cells  –  purple
 Pituitary βcells  –  purple
 Sulphated mucins  –  purple
 Background  –  as the counterstain
 Nuclei  –  as the nuclear stain

Tricromica de gomori

Se trata de una solución acética estabilizada que contiene Chromotrope 2R, Azul de Anilina
y Acido Fosfotúngstico. Listo para usar. No requiere dilución.
Colorante Histológico. Tiñe citoplasma y fibras musculares de rojo, mientras que el
colágeno (protaminas) toma un color azulado.

Phosphotungstic acid is used in histology as a component for staining of cell specimens,

often together with haematoxylin as PTAH. It binds to fibrin, collagen, and fibres of
connective tissues, and replaces the anions of dyes from these materials, selectively
decoloring them.
Cromotropo 2R: A red acid dye, C16H10N2O8S2Na2, used as a counterstain and for
staining red blood cells in sections.

Tinción de Bielschowsky: coloración de neurofibrillas y cilindroejes con plata amoniacal.

Los axones se tiñen de negro y los cuerpos neuronales toman un color dorado oscuro
PRINCIPLE: The nerve fibers are sensitized with a silver solution. The
sections are treated with ammoniacal silver, and then reduced to a visible
metallic silver.

PROCEDURE:
 1. Hydrate to distilled water, 3 changes.
2. Preheat a coplin jar of 20% Silver nitrate solution, 35 seconds in the
microwave, add slides, place in a 37°C oven for 15 minutes.
3. Wash in distilled water, 3 changes.
4. Place slides in Ammoniacal silver solution, 10 minutes, in 37°C oven.
5. Place slides in Working Developer solution, 1 to 5 minutes, check
under microscope.
6. Wash in fresh ammonia water.
7. Wash in distilled water, 3 changes. 8. Place slides in 5% Hypo, 2 minutes.
9. Wash in distilled water, 3 changes.
10. Dehydrate, clear and mount.
RESULTS:
axons, plaque neurites and tangles: black
background: yellow to brown
plaque and vascular amyloid: generally brown to dark brown
Luxol fast blue

Any of a group of closely related copper phthalocyanine dyes used as stains for myelin in
nerve fibers
Luxol fast blue is commonly used to observe myelin under light microscopy. It is the
alcohol soluble counterpart of the water soluble alcian blue. The stain works via an acid-
base reaction with the base of the lipoprotein in myelin replacing the base of the dye and
causing a colour change.
Under the stain, myelin fibres appear blue, neutrophil appears pink, and nerve cells appear
purple
PRINCIPLE: Luxol Fast Blue is the alcohol soluble counterpart of the
water soluble Alcian Blue. Staining is due to lipoproteins, and the
mechanism is one of an acid-base reaction with salt formation; the base
of the lipoprotein replaces the base of the dye.
Tinta china
En ocasiones, las partículas relativamente grandes que estas células fagocitan oscurecen su
núcleo, pero incluso cuando este no puede distinguirse, la presencia de partículas
extrañasdentro de una célula bastante grande indica, por lo general, que se trata de cierto
tipo de macrofago. Las partículas engullidas, incluyendo marcadores experimentales (tinta
china o azul de tripano), pasan al citoplasma en fagosomas que después se unen a los
lisosomas primarios. Las enzimas hidroliticas de estos ultomos atacan y degradan a las
macromoléculas orgánicas de material ingerido.
Colorantes de sales de plata (los llamados de impregnación argéntica) son útiles para teñir
el cuerpo y las ramificaciones neuronales (dendritas y axón), visibles al microscopio de luz
o electrónico. Estos métodos, conocidos como de Golgi (en honor a su descubridor, el
anatomista italiano Camilo Golgi, a finales del siglo pasado), representaron un avance
cualitativo de nuestro conocimiento acerca de la morfología del SNC, y en particular,
gracias a los estudios de Santiago Ramón y Cajal, en España. No todas las neuronas se
tiñen con estas sales de plata, y sigue siendo un misterio por qué sólo algunas de estas
células son afines al colorante.
Certain tissue components called argyrphilic have a natural affinity for silver salts. The
glycoproteins in these materials reduce silver salts to silver metal, depositing a black stain
around the argyrophilic materials. Reticular fibers and the granules in diffuse endocrine
cells are argyrophilic. Usually a counterstain is not used and the unstained cellular elements
are seen as colorless shadows.
Silver complexes employed to demonstrate reticular fibers in normal and diseased tissues,
as well as neuroglia, neurofibrillae, argentaffin cells, and Golgi apparatus

Las soluciones de hematoxilina férrica se utilizan especialmente para tinciones nucleares

en cortes tisulares en los que se contratiñe con soluciones colorantes ácidas. Ejemplos son
la representación de fibras elásticas según van Gieson y las diferentes tinciones tricrómicas
del tejido conjuntivo. Esto se debe a la elevada resistencia de la tinción con hematoxilina
férrica frente a las soluciones colorantes ácidas. Las soluciones usuales de hemalumbre
como hemalumbre de Mayer, Delafield, Gill, Harris, dan en combinación con
contratinciones ácidas solamente tinciones nucleares débiles.
Staining with iron hematoxylin solution produces selective nuclei staining that is stable and
requires no additional differentiation. This type of staining is more resistant to acid staining
solutions than conventional hemalum solutions. Weigert’s iron hematoxylin staining is all the more
necessary when dealing with decalcified material or with stains where acid solution has been used
for counterstaining, e.g. in visualising elastic fibres according to van Gieson, or in the case of
various trichrome connective tissue stains acc. to Gomori, Masson, Wallart & Houette or Goldner.

Result
Nuclei blue-black

Tricrómica de Pollack
INGREDIENT:
ETHANOL (C2H5OH)
METHANOL CH3OH)
ACID FUCHSIN (C20H17N3NA2O9S3)
LIGHT GREEN SF (C37H34N2NA2O9S3)
ORANGE G (C16H10N2NA2O7S2)
PHOSPHOTUNGSTIC ACID (NA3PW12O4OXH2O)
GLACIAL ACETIC ACID (CH3COOH)
PONCEAU 2R (C18H14N2O7S9NA9)

Acid Fuchsin is one of the dyes used in Masson's Trichrome Stain. The muscle stains red with the
acid fuchsin, and the collagen is stained green or blue with light green SF yellowish or methyl blue.
Xylidine ponceau is mostly used only in the Masson's trichrome stain as a red counterstain, where
it imparts orange hue to the red-stained cytoplasmic structures.
The main use of Orange G is in the OG6 Papanicolaou stain, to stain keratin. It is often combined
with other yellow dyes and used to stain erythrocytes in the trichrome methods.