The excess stain is removed with dilute hydrochloric acid solution acid water. The effect of the staining technique on morphological and. The oxidation product of haematoxylin is haematin, and. A loopful of curd is placed on a clean grease free slide and diluted with 1 drop of sterilised water. Papanicolaou staining method is the routine staining procedure used in cytopathology. Spread the sputum evenly over the central area of the slide using a continuous rotational movement. Some staining techniques, such as the gram and ziel neelsen stains, although of great diagnostic value because of their differential staining properties for specific bacteria, reveal little. In papanicolaou stain, fixed smears are passed through a series of descending grade of ethyl alcohol before nuclear staining. A manual for cytotechnology, c keebler, j reagen, editors, american society of. Negative staining, simple staining, differential staining, and structural staining. The purpose of capsule staining is to observe bacterial capsule by distinguishing capsular material from the bacteria cell.
This article follows on from nicolas introduction, but well take an indepth look at the stains, chemistry and method to enable you to perform this to high standards in your. Gently rinse off the stain with water and shake off the excess. Introduction to production part approval process ppap production part approval process ppap defines generic requirements for production part approval. Acidfast bacteria, of which there are very fewthe major genus mycobacterium, have a high concentration of mycolic acid, a lipid, in their walls. How to perform capsule staining of a bacteria experiment. The document also includes the 9 standard operating procedures for the pap staining in cervical. Exfoliative cytology is a simple, noninvasive diagnostic technique which could provide as an adjunct in early. Objectives to learn the technique of smear preparation. Introduction quality assurance many of the stains that are described in this national standard method nsm are commercially available. The decolourising process is then stopped by immersing the slide in running tap water.
Haematoxylin and eosin staining is the most common staining in the modern and old. Hematoxylin staining requires the use of a mordant most commonly aluminum salts and stains the nuclear components of cells a dark blue. Guidelines of the papanicolaou society of cytopathology. Although difficult to stain, once the stain goes into the wall, the cell will not destain or decolorize easily. In capsule staining, a smear of bacteria is made at the center of a slide. Metaplastic cells often stain both green and pink at once. It is not heatfixed, as the cell shrinkage caused by heating may create a clear zone around the cell, which may be mistaken as capsule.
Basic fuchsin binds to lipids in the cell wall and turns mycobacteria red. This staining technique gives an overview of the structure of the tissue and can be used in pathological diagnosis. Although we publish a procedure for the pap stain, individual laboratories may need, because of past experience and what stain picture the technicians are used to, have a trial approach to the procedure and make adjustments. This preparation method is useful for visualizing suspensions of small particles, which includes viruses, purified proteins, liposomes and small vesicle fractions. The polychromatic pap stain involves five dyes in three solutions. Do not allow the staining solution to dry on the slides add fresh solution as required. Hematoxylin is used in combination with eosin because eosin stains the cytoplasmic organelles varying shades of pink, red or orange. Papanicolaou society of cytopathology fineneedle aspiration fna is a simple, safe, and costeffective procedure for the investigation of patients with a mass. A pap smear involves collecting cells from your cervix the lower, narrow end of your uterus thats at the top of your vagina. The purpose of ppap is to determine if all of alto shaams engineering design records and specification requirements are properly understood by the supplier, and that the. Production part approval processppap production and part approval process ppap page 410 release. Cytoplasmic staining with counterstins orange g and ea 4. The gram stain procedure was originally developed by the danish physician hans christian gram to differentiate pneumococci from klebsiella pneumonia. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic staining histological technique developed by george papanikolaou, the father of cytopathology.
In brief, the procedure involves the application of a solution of iodine potassium iodide to cells previously stained with crystal violet or gentian violet. After haematoxylin staining in the pap stain procedure, og6 is the first counterstain to be used and must be. Flood airdried, heatfixed smear of cells for 1 minute with crystal violet staining reagent. Deparaffinize sections by sequential immersion in the following for 2 minutes each. Place slides containing paraffin sections in a slide holder glass or metal deparaffinize and rehydrate sections.
Rinse very carefully with several exchanges of deionized water. Petri dishes should be covered throughout the staining procedure to provide a humidified atmosphere. Introduction to production part approval process ppap. In the first step, the nuclei are stained with a hematoxylin solution. As such, it is used for cervical cancer screening in gynecology. No known test method can offer complete assurance that blood samples. Original article comparison of the routine papanicolaou. The papanicolaou stain is one of the most widely used stains in cytology, where it is used to aid pathologists in making a diagnosis. The bv smears should be packaged so as to protect the glass slides from breaking en route, and maintain ambient temperature during shipping. Rapid economic, acetic acid, papanicolaou stain ihc world. It has affinity for chromatin, attaching to sulphate groups on the d.
Staining methods histology and cytology 152 notes sources of errors irregular distribution of the blood smear on a glass slide may result in an erroneous cell counts. Pdf microcracks are one of the determinants of the bone strength and their accumulation may contribute to increased fracture risk. The pas staining procedure may also be used for the demonstration of fungal organisms in tissue sections. Module common staining technique microbiology 24 notes indeterminate groups as well. Laboratory manual for the examination and processing of human semen. Cells with high content of keratin are yellow, glycogen stains yellow as well. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic multicolored cytological staining technique developed by george papanicolaou in 1942. The slides have to be examined at different interval to find out when the staining is optimum. These ethyl alcohol grades are replaced by single 1% acetic acid step.
Papanicolaou staining is the universal method for staining gynecologic slides. Place a ring around the section using a pap pen khp001 or equivalent method to prevent the spreading of reagents on the slide. Papanicolaou stain is a multichromatic multicolored cytological staining technique developed. In the acidfast staining procedure, mycobacteria are first stained with the red dye basic fuchsin and the cells are then decolorized and restained with methylene blue. Do not let the specimens dry at any stage of the staining procedure. Papanicolaou stain includes both acidic and basic dyes. Modified method of agnor staining and interpretation was an easy, reliable and reproducible alternative to traditional agnor techniques for evaluating proliferation activity of cells in malignant and benign brain lesions. The word gram is always spelled with a capital, referring to hans christian gram, the inventor of gram staining gram staining principles gram staining is used to determine gram status to classify bacteria broadly. Pap staining is used to differentiate cells in smear preparations of various bodily secretions. Regressive staining in this method the tissue is overstained and then destained differentiate until the proper endpoint is reached. A thin smear is then prepared on the slide, dried in air and finally heat fixed. The smear is stained following the usual procedure of gram staining and examined under microscope using oil.
Prepared by roy ellis imvs division of pathology the queen elizabeth hospital woodville road, woodville, south australia 5011 novaultra special stain kits principle. This procedure specifies the requirement of quality assurance for identification of materials during the manufacturing process. Superficial cells are orange to pink, and intermediate and parabasal cells are turquoise green to blue. If the slide is heat fixed, allow it to cool to the touch before applying the stain. Gluteraldehyde should be avoided as a tissue fixative since it is a dialdehyde that will react with this reagent and give falsepositive pas staining. Negative staining is a simple technique for routine examination of structure. Add periodic acid solution to staining dish for 10 minutes. Acidic dye stains the basic components of the cell and basic dye stain the acidic components of the cell. Terms in this set 19 4 main steps of the pap stain. Periodic acid schiff pas reagents are for in vitro diagnostic use. Fine needle aspiration was carried out from various organs as an outpatient procedure for patients referred from different clinical departments for diagnostic purpose.
Pdf staining procedure for the detection of microcracks. Progessive staining when tissue is left in the stain just long enough to reach the proper end point. Total time for staining in the present method is 3 minutes c. Please note that the quality of the smear too heavy or too light cell concentration will affect the gram stain results.
If using xylene substitutes, follow the directions provided by the. Place slides on dryer with smeared surface upwards, and air dry for about 30 minutes. Staining procedures scope of document this test procedure contains the methods for the staining procedures commonly used in clinical microbiology laboratories for the identification of pathogens. Run 95% ethyl alcohol down the slide until the solvent runs clear about 1020. Periodic acidschiff pas staining system procedure no. If observed results vary from expected results, please contact sigmaaldrich technical service for assistance. The components to be tested are sfc valve interlocks. The staining procedure should clearly define the details of all structures. Once the slides have arrived at the contractors facility, they will be logged in, heat fixed and stained as described. Og6 is a reliable stain that will perform through the lifetime of the product. It is to measure the concentration of elements without causing any.
Natural dye hematoxylin is the nuclear stain which stains cell nuclei blue. Using forceps, tilt each slide to drain off the staining solution. Procedure of regressive papanicolaou staining method. Download file international journal of contemporary medical.
Also known as the pap test, pap smear and smear test a pap test is a microscopic examination of cells that are scraped from a mucosal surface after being stained with papanicolaou stain. A pap smear, also called a pap test, is a procedure to test for cervical cancer in women. The sigmaaldrich papanicolaou staining system is intended for staining exfoliative. Papanicolaou stain pap stain protocol ihc world, llc. Extraction slide will have no visible pas staining of glycogen when compared to the undigested glycogen positive control slide. When using the regressive staining method, the nucleus is deliberately overstained with a nonacidified haematoxylin. The recommended size of the smear is about 20 mm by 10 mm. Comparison of modified ultrafast papanicolaou stain with the standard rapid papanicolaou stain in cytology of various organs. Alcohols used for wiping the skin may cause hemolysis and artifacts. Comparison of modified ultrafast papanicolaou stain with.
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