Procedure
- Prepare a film of blood or bone marrow on a microscopic slide and allow to air dry.
- Place the air-dried smear on the slide staining rack, smear side facing upwards.
- Cover the blood film with undiluted staining solution. The undiluted stain fixes and partially stains the smear.
- Let stand for 2-3 minutes.
- Add approximately equal amount of buffered water (pH 6.5). ...
- Leave for 5 minutes.
Full Answer
How do you stain a blood smear?
Blood smear staining. Procedure • Thin blood films (only) – Dip Method 1. Dip air dried blood film in undiluted stain for 15 to 30 seconds (double the staining time for bone marrow smears). 2. Decolorize the stained smears by immersion in distilled or deionized water and air dry 3. Let air dry in a vertical position.
How long does it take to stain blood film?
Dip air dried blood film in undiluted stain for 15 to 30 seconds (double the staining time for bone marrow smears). 7. Decolorize the stained smears by immersion in distilled or deionized water and air dry 8.
How do you draw blood from a drop of blood?
Slide Method Procedure Place a drop of blood in the centre of a clean glass slide 1 to 2 cm from one end. Place another slide (spreader) with smooth edge at an angle of 30-45⁰ near the drop of blood. Move the spreader backward so that it makes contact with drop of blood.
What is the purpose of preparing blood smear (blood film) and staining?
Preparation Blood Smear (blood film) & Staining.... The purpose of preparation of blood smear (blood film} is to study the morphology of RBCs, differential leukocyte count and reticulocyte count.
How do you stain blood slides?
2:145:22Blood Smear Preparation and Staining Practical Lab - YouTubeYouTubeStart of suggested clipEnd of suggested clipAlternatively you may fill a Copland jar with Leishman stain and immerse the slide in the stain.MoreAlternatively you may fill a Copland jar with Leishman stain and immerse the slide in the stain. Staying for about one minute and then add twice the volume of distilled water to the stain.
What stain is used to stain blood smear?
Blood films are routinely stained with a Romanowsky-type stain (e.g., Wright or Wright-Giemsa) either manually or using an automatic slide stainer. Romanowsky-type stains are composed of a mixture of eosin and oxidized methylene blue (azure) dyes.
How we prepare and stain the blood film?
StainingPrepare fresh working Giemsa stain in a staining jar, according to the directions above. ... Pour 40 ml of working Giemsa buffer into a second staining jar. ... Place slides into the working Giemsa stain (2.5%) for 45-60 minutes.Remove thin smear slides and rinse by dipping 3-4 times in the Giemsa buffer.More items...
How do you stain slides?
0:000:56How to Stain a Slide - YouTubeYouTubeStart of suggested clipEnd of suggested clipWe're going to put a drop on one side. And it wicks over just a little bit. But if we take the paperMoreWe're going to put a drop on one side. And it wicks over just a little bit. But if we take the paper towel and put it on the other side whoa. It basically sucks the stain across the slide.
Why do we stain blood smears?
The purpose of examining a blood smear is to check the size, shape, and number of three types of blood cells: Red blood cells, which carry oxygen from your lungs to the rest of your body. White blood cells, which fight infection. Platelets, which help your blood to clot.
Can you Gram stain a blood smear?
Gram stains may also be used to check for bacteria in certain body fluids, such as blood or urine. There are two main categories of bacterial infections: Gram-positive and Gram-negative. The categories are diagnosed based on the how the bacteria reacts to the Gram stain.
How do you prepare a blood smear slide?
A thin smear being prepared. Place a small drop of blood on the pre-cleaned, labeled slide, near its frosted end. Bring another slide at a 30-45° angle up to the drop, allowing the drop to spread along the contact line of the 2 slides. Quickly push the upper (spreader) slide toward the unfrosted end of the lower slide.
How do you stain thick blood film?
For thick film preparations a small drop of blood is placed on a glass slide and spread to approximately four times its original surface. After extensive drying, best done at 50 to 60°C for 7 to 10 minutes, the slides can be stained. The cells wash off the slide if insufficiently dried.
Why are stains used while preparing slides?
Staining is done to visualise the specimen clearly under the microscope. Without staining the specimen under the microscope will look transparent or will not be visible clearly. Hence, staining is done to make the components of cell clearly visible.
What stain is used for slides?
Microscope Slide Stains Stains such as methylene blue in low concentrations does not harm the tissues and so can be safely used on living materials. Such stains are called vital stains. For making temporary slides stains such as methylene blue, idodine, aniline hydrochloride, safranin etc are used.
How do vets stain slides?
The staining procedure: Make sure the sample on the slide is dry. Dip the slide in each jar between six-10 times (10-15 seconds in each solution). ... Wash the slide with water after jar number 3 ONLY. Dip the slide in the water jar. Never wash the slide in between the staining process. Alternative to Jar 4:More items...
What can I use to stain microscope slides?
There are several types of stains you can use at home including: food coloring, iodine, malachite green (ick fish cure), and methylene blue. You can purchase food coloring at a grocery store, iodine at a pharmacy, and the malachite green and methylene blue can be purchased at an aquarium store.
What is Leishman stain used for?
Leishman stain, also known as Leishman's stain, is used in microscopy for staining blood smears. It is generally used to differentiate between and identify white blood cells, malaria parasites, and trypanosomas.
How do you stain a blood smear with Wright's stain?
ProcedurePrepare a film of blood or bone marrow on a microscopic slide and allow to air dry.Place the air-dried smear on the slide staining rack, smear side facing upwards.Cover the blood film with undiluted staining solution. ... Let stand for 2-3 minutes.Add approximately equal amount of buffered water (pH 6.5).More items...•
How do you stain a blood smear with Giemsa?
MethodDeparaffinize and rehydrate through graded alcohols to water.Rinse in pH 6.8 buffered distilled water.Stain in working Giemsa, overnight.Rinse in distilled water.Rinse in 0.5% aqueous acetic acid until section is pink.Wash in tap water.Blot until almost dry.
What is Wright Giemsa stain used for?
Wright-Giemsa Stain Kit ab245888 is intended to be used for differential staining of blood smears, bone marrow and blood parasites. Neutrophils: Light Purple or Lavender granules in cytoplasm.
How long to let Jenners stain stand?
HorizontalStaining Protocol: • Place slide with thoroughly dried film in a horizontal staining rack • Flood smear with absolute methanol for 15-30 seconds and then drain • Flood smear with 1ml Jenners Stain Solution and let stand for 3 minutes • Add 1mL of pH 6.6 Phosphate Buffer solution and 1 ml deionized water to smear and let stand for 45 seconds • Rinse briefly with running deionized water • Air dry and examine under a microscope • Perform immuno chemical staining procedure according to manufacturer.
How to use Wright's stain?
Procedure • Thin blood films (only) – Dip Method 1. Dip air dried blood film in undiluted stain for 15 to 30 seconds (double the staining time for bone marrow smears). 2. Decolorize the stained smears by immersion in distilled or deionized water and air dry 3. Let air dry in a vertical position. • Thin blood films (only) – Rack Method 1. Lay air dried slides on staining rack and flood with stain; stain for 10 to 15 seconds (double the staining time for bone marrow smears). 2. Add an equal volume of deionized/distilled water and stain for 10 seconds. 3. Rinse the slide by dipping in deionized/distilled water for 30 seconds.The slide may also be rinsed by swishing or washing with deionized/distilled water.
How to make Leishman stain?
Leishman Stain: Preparation • Dissolve 0.2 g of powdered Leishman’s dye in 100 ml of acetone-free methyl alcohol in a conical flask. • Warm it to 50°C for half an hour with occasional shaking. • Cool it and filter it. Procedure for staining • Pour Leishman’s stain dropwise (counting the drops) on the slide and wait for 2 minutes. This allows fixation of the PBF in methyl alcohol. • Add double the quantity of buffered water dropwise over the slide (i.e. double the number of drops). • Mix by rocking for 8 minutes. • Wash in water for 1 to 2 minutes. • Dry in air and examine under oil immersion lens of the microscope.
What are the ingredients in Romanowsky stains?
All Romanowsky combinations have two essential ingredients i.e. methylene blue and eosin or azure. • Methylene blue is the basic dye and has affinity for acidic component of the cell (i.e. nucleus) and eosin/azure is the acidic dye and has affinity for basic component of cell (i.e. cytoplasm). • Most Romanowsky stains are prepared in methyl alcohol so that they combine fixation and staining.
What are the two types of blood film?
4. The peripheral blood film (PBF) is of two types: 1. Thin blood film 2. Thick blood film Three basic steps to make blood film: 1. Preparation of blood smear. 2. Fixation of blood smear. 3. Staining of blood smear.
What is the aim of blood smear?
Aim of blood smear • Blood films are usually examined to investigate hematological problems (disorders of the blood) and , occasionally , to look for parasites within the blood such as malaria and filaria.
Why is blood film important?
• Examination of thin blood films is important in the investigation and management of anaemia, infections , and other conditions which produce changes in the appearance of blood cells and differential white cell count. • A blood film report can provide rapidly and at low cost, useful information about a patient’s condition.
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 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 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 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.
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 slide oil immersion lens?
slide and Shift the Oil immersion lens by constantly looking from the side of the microscope so that it just dips in the oil.
How long to stain slides with Giemsa stain?
Place slides into the working Giemsa stain (2.5%) for 45-60 minutes.
How long after blood draw should you do a thin smear?
Make as many thin smears as possible, preferably within one hour after the blood was drawn from the patient.
What is the Wright-Giemsa stain used for?
Wright (Wright-Giemsa) stain. Used in hematology, this stain is not optimal for blood parasites. It can be used if rapid results are needed, but should be followed up when possible with a confirmatory Giemsa stain, so that Schüffner’s dots can be demonstrated.
Can you stain a smear of malaria?
Just before use, remove the smear from the box and allow the condensation to evaporate; label the slide “+ malaria” and the present date. The smear is now ready for staining since it was previously fixed.
Can you stain blood smears?
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.
Do you need a positive smear for a Giemsa stain?
To ensure that proper staining results have been achieved, a positive smear (malaria) should be included with each new batch of working Giemsa stain. Since good quality control smears are not available commercially, they may be prepared from a patient’s blood and stored for future use in the following manner:
What is slide staining?
Microscope slide staining is a microscopy technique used to help study microscopic objects that are difficult to see clearly, even at high magnification. By introducing a dye to a specimen on your slide, you can visually contrast your specimen and the surroundings.
Why is it important to use a microscope slide stain?
However, sometimes that isn’t enough, as many objects and microscopic organisms are naturally translucent. Microscope slide staining overcomes this obstacle and further enhances our experience, so we can observe these microscopic structures in much greater detail. Although staining is an important microscopy technique, the chemicals involved can be quite dangerous so be sure to use all the proper precautions if you intend to use them.
What Are the Dyes Used in Microscope Slide Staining?
Some dyes are generally used to be absorbed by the subjects in a sample, as with bright-field microscopy.
What are slide dyes used for?
Types of Slide Dyes. There are many dyes that are used with a microscope slide staining. Some of them are used for live cells, whereas others are designed for brightening purposes. If you want to know which dyes you can use with your microscope, you’re in the right place.
How do dyes get their color?
Most dyes are types of salt and gain their color from colored ions or chromophores. The way a dye reacts with a substance, whether it is absorbed completely, partially, or not at all, depends on the chemical makeup of both the substance and the dye.
How long to let bunsen slide sit?
With the paper still on top, the slide is held over a bunsen burner for five minutes. The paper is removed, and the slide is carefully rinsed with water. The second stain is applied and left to sit for two minutes. The slide is carefully rinsed again and then blotted dry.
How long to hold a paper slide over a bunsen burner?
With the paper still on top, the slide is held over a bunsen burner for five minutes.
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.
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 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.
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.
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, ...
What is the angle of the spreader slide?
This makes the blood spread along the base of the slide. The smear is made with the spreader inclined at an angle of approximately 30° to the blood.
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.
How to draw a stain on a slide?
The image above shows how to draw a stain into a prepared slide. With the cover slip in place on top of the specimen, place a drop of stain on the edge of the cover slip. On the opposide side of the cover slip place a paper towel or cloth to draw the liquid out from the cover slip.
What is slide staining?
Microscope Slide Staining Information. Microscope cell staining is a technique used to enable better visualization of cells and cell parts under the microscope. By using different stains, a nucleus or a cell wall are easier to view. Most stains can be used on non-living (fixed) cells, while only some types of stain can be used on living cells.
What color is used in gel electrophoresis?
Coomassie Blue - stains proteins a bright blue, and is often used in gel electrophoresis. Crystal Violet - stains cell walls purple when combined with mordant. This stain is used in Gram Staining. DAPI - a fluorescent nuclear stain that is excited by ultraviolet light, showing blue fluorescence when bound to DNA. LIVE.
Why do we use stain on cells?
Cells are primarily stained to enhance visualization of the cell or certain componenets . Cells are sometimes also stained to highlight metabolic processes or to differentiate between live and dead cells.
What color is iodine?
Iodine - used as a starch indicator. When in a solution, starch and iodine turn a dark blue in color. Malachite Green - a blue-green counterstain to safranin in Gimenez staining for bacteria. This stain is often used to stain spores.
What color is ethidium bromide?
Ethidium Bromide - this stain colors unhealthy cells in the final stages of apoptosis, or deliberate cell death, fluorescent red-orange.
Can stain be used on cells?
Below is a list of commonly used stains, often for different types of cells. All those listed can be used on fixed (non-living cells) and any that can be used on living cells are noted at the end of the description with the word "LIVE".
What to do when looking at blood smears?
When you look at a blood smear, it’s best to have a plan, and it’s best to try to follow it each time. It might sound boring – but you’ll make a much more accurate and complete assessment that way. Otherwise, the temptation is to just put the slide under the microscope, scan around to see if you see anything weird, ...
What to evaluate on a blood smear?
1. Red blood cell number. First, make sure you’re in the right part of the smear.
