What is the purpose of staining blood smear?
Staining Blood Smears. Stain only one set of smears, and leave the duplicates unstained. The latter will prove useful if a problem occurs during the staining and/or if you wish later to send the smears to a reference laboratory.
What are the different types of staining?
They include: 1 Giemsa stain 2 Wright and Wright-Giemsa stain 3 May-Grunwald stain 4 Leishman stain
What is malaria stain used to test for?
This is a special stain used for examination of blood films for parasitic infections and majorly for the diagnosis of malaria. It is also used as a differential stain for various blood cells (erythrocytes, platelets, leucocytes) and cellular components such as the nuclear and the cytoplasm.
What stain gives the best nuclear and cytoplasmic detail?
Romanowsky-type stains give good nuclear and cytoplasmic detail. Red blood cells stain red-orange, nuclei stain blue-purple and cytoplasm stains blue to pink. Most commercial laboratories use some form of Romanowsky-type stain (e.g. Wright - Geimsa) and these stains give excellent result but tend to be fussy.
How long does it take for a smear to stain?
The dried smear is then fixed with methanol or ethyl alcohol and stained. The smear is covered with stain for approximately ten minutes, then diluted with water and allowed an additional ten minutes for the cells to properly stain. Following the stain application, the slide is rinsed under running water.
Why is a blood smear test important?
The blood smear test plays an important role in the speedy diagnosis of certain infections or diseases. This test uses a drop of blood spread onto a glass microscope slide that is then treated with a colored stain and examined using a microscope. The blood smear test shows a sample of blood components including platelets, ...
How to tell if a smear is too thin?
At least two slides should be made during testing. A smear will be too thin if the spreader slide is moved too quickly or if the angle of the spreader is less than 30°. Conversely, the smear will be too thick if the spreader is moved slowly or if the angle is greater than 30°. Large blood drops may extend the smear over too much of the base slide, while a small drop can be insufficient for the smear. The stain needs adequate time with the sample to avoid over-staining or under-staining. If a sample is over-stained, debris might show up in the sample. This can also happen if the stain is not washed enough with running water.
What is the slide called that is used to spread blood?
Another microscope slide called a spreader slide is used. This slide should have chipped edges, along with another smooth end. The side of the spreader slide with chipped edges is placed on the original slide (base slide) in front of the blood and moved backwards to touch the blood. This makes the blood spread along the base of the slide.
How is a smear made?
The smear is made with the spreader inclined at an angle of approximately 30° to the blood. The smear should cover two-thirds of the base slide and should have a feathered end. The smear should then be air dried. The frosted end of the slide should be labeled with the patient's name, identification number, and date. The dried smear is then fixed with methanol or ethyl alcohol and stained.
What color are reticulocytes?
Blood cells stain in their center. Reticulocytes are immature blood cells, and they stain a darker blue color, while mature erythrocytes stain a lighter blue color. Staining intensity varies according to the stain concentration and the sample's contact time with the stain. Interpretation.
What is the Wright stain?
The Wright stain, also known as the Romanowsky stain, is a mix of both acidic and basic dyes that are used to distinguish cellular components.
What is the purpose of blood smear?
The purpose of preparation of blood smear (blood film} is to study the morphology of RBCs, differential leukocyte count and reticulocyte count.
How to add Leisman's stain to a smear?
Add Leisman’s stain drop by drop till it covers whole of the smear. Count the number of drops you have put.
What is the color of Leishman's stain?
1.5 gm powder of Leishman’s stain is dissolved in one litre of acetone free methyl alcohol. Leishman’s stain contains two dyes, eosin and methylene blue. Eosin is an acidic dye that stains basic structures like RBC and granules of eosinophil. It is pink or red in colour. Methylene blue is a basic dye that stains acidic structures like nucleus or granules of basophils. It is blue in colour. Acetone free methyl alcohol is a fixative for smear.
How to get pink smear out of slide?
Wash the smear in slow running tap water or with the help of wash bottle till the smear becomes pink in colour. Clean the back of the slide to remove the stain from back side.
How long does it take for a stain to dry?
Wait for 8-10 minutes for staining to complete. During this period in the presence of buffered water staining is taking place because of formation of cations and anions of basic and acidic dyes respectively. Methyl alcohol is unable to ionise the stain so unable to stain the cells.
What is the shape of a smear?
It is tongue shaped having head, body and tail. Head is the area where blood drop is placed. Body is the area between head and tall. Tail is the last part of the smear with ragged margins.
Why is a smear fixed?
Fixation of smear is because of precipitation of proteins by alcohol which prevent washing off of the film.
Why is chromosome staining used?
It is used to stain chromosomes to diagnose various diseases, infections, and syndromes such as leukemia.
How to clean a smear of a smear?
Using water, rinse the stained smear, ensuring the pH level of the rinsing agent is 6.5. Thoroughly rinse the stained smear until the edges of the smear turn light pinkish-red. The underside may have some water stains; wipe it gently, and air dry in a vertical position. Check the slide under the microscope.
What is the principle of Wright’s Stain?
It is a staining procedure polychromatic in nature consist of Methylene blue and eosin mixture.
What are the reagents used?
You can buy the dye in powder form and mix it to methanol or readily available solutions.
What are the differences between Wright’s stain and Giemsa Stain?
Although Wright’s stain and Giemsa stain are both differential staining and alike in many ways, they have differences.
What color is used in Romanowsky stain?
Since it is derived from Romanowsky stain, you can expect to use the typical components like eosin Y, oxidized methylene blue , and azure B dyes.
How long does Wright's stain last?
Wright’s stain can last for up to 52 weeks from the manufacturing date provided stored the right way.
What is the best way to make a blood smear?
Always start with room temperature, well-mixed, clot free, EDTA anticoagulated blood (heparin is used for some exotic species).
What color is the cytoplasm stain?
Romanowsky-type stains give good nuclear and cytoplasmic detail. Red blood cells stain red-orange, nuclei stain blue-purple and cytoplasm stains blue to pink.
What are the problems with rapid staining?
The most common problems arising from use of rapid stain are 1) poor sample fixation 2) under-staining of the specimen 3) poor staining from use of weakened or exhausted stain or fixative and 4) stain contamination 5) excessive stain precipitation. Poor sample fixation.
What are the lines on a smear called?
Often there is streaking in the smear. Linear lines arranged horizontally to the leading edge are called hesitation marks and they indicate hesitation in the forward motion. Excessive downward pressure will produce short slides with hesitation mark s and a poorly developed feather edge and monolayer region.
How long should a cytology smear be fixed?
For very thin cytology smears and for blood smears, 5 or 6 dips in the fixative solution should be adequate, but for thick cytology smears, up to 120 seconds may be required for adequate fixation. When in doubt, fix them a little longer. Under-staining of the specimen.
Do slides need to be dried before staining?
Slides should be completely dry before staining (use of a gentle amount of warm air from a hair dryer can facilitate this), slide holders should be dried before slides are loaded, and stains should be tightly covered when not in use. Stain precipitation.
Is polychromasia semi-quantified?
Polychromasia is typically semi-quantified as rare, slight, mild, moderate or marked (can use similar criteria as with morphology quantification, see below). The transition zone between the feathered edge on the right and the monolayer on the left.
What are the three types of blood stain?
They include: Giemsa stain. Wright and Wright-Giemsa stain. May-Grunwald stain. Leishman stain. The complex of these stains was coined after Dmitri Leonidovich Romanowsky, a Russian Physician who first identified the importance of using blood samples to make and identify effects using blood staining methodologies.
What are stains made of?
The stains are neutral, made up of oxidized methylene blue (azure) dyes and Eosin Y. The azures are basic dyes that bind to the acid nuclei forming a blue-purple color. The acid dye, Eosin binds to the alkaline cytoplasm forming red coloration.
What are the Romanowsky Stains?
Romanowsky Stains are the stains that are used in hematology and cytological studies, to differentiate cells in microscopic examinations of blood and bone marrow samples. These stains are also applied to detect the presence of parasites in the blood such as malaria parasites. There are various Romanowsky staining types that apply the same principle. They include:
What is the May-Grünwald-Giemsa stain?
May-Grünwald-Giemsa stain. It is a two-step staining procedure whereby the first staining is done with May-Grünwald stain and a second stain of Giemsa stain which produces the Romanowsky effect (wide range of hue/color)
What is the color of eosin Y?
The mixture of active eosin Y and active methylene blue was attributed to the formation of hues that distinguished the cell components, in that shades of purple, are formed in the cell chromatins of the nucleus and the granules in the cytoplasm of some white blood cells, to which Romanowsky defined as the Romanowsky effect or Romanowky-Giemsa effect.
What color is the Wright's stain?
When Giemsa stain is added to the Wright’s stain, the color brightens to a reddish-purple in the cytoplasmic granules. These are hematological stains that are used to differentiate blood cells from peripheral blood smears, bone marrow aspirates, and urine samples.
What is the name of the stain that is a combination of Wright stain and Giemsa sta?
A combination of Wright stain and Giemsa stain is known as the Wright-Giemsa stain.
