This technique was developed by george papanicolaou, the father of cytopathology. Inception and development of the papanicolaou stain method. Principles of pap staining multichromatic staining technique classic form. Papanicolaou stain papanikolow, a multichromatic stain used principally on exfoliated cytologic specimens and based on aqueous hematoxylin with multiple counterstaining dyes in 95% ethyl alcohol, giving great transparency and delicacy of detail.
Scribd is the worlds largest social reading and publishing site. The modification is achieved by increasing the staining time for the three basic papanicolaou stains and incorporating phloxine b as an additional staining component. Some bacteria have nonionic charge or slime layer on their surface. Papanicolaou stain also papanicolaou s stain and pap stain is a multichromatic staining histological technique developed by george papanikolaou, the father of cytopathology. The following image provides an example of ihc staining. The intensity of the nuclear staining is controlled by the immersion of the slide into a blueing agent. Do not let the specimens dry at any stage of the staining procedure. Negative staining procedure microbiology with sumi youtube.
Full text full text is available as a scanned copy of the original print version. The 5dye pap stain method evolved through 2 salient phases. A syringe piston handle can be used, leaving one hand free to immobilize the lesion. The technique of washing tissue specimens with a concentrated solution of a heavy metal salt and letting it dry. Pap staining procedure was carried out approximately in 300 seconds with procedures as per. Staining paraffin sections by pap procedure pdf mouse monoclonal antibodies procedure. Prepare bacterial smear on clean and grease free slide, using sterile technique. Semen smear fixed in 95% reagent alcohol or cytology fixative.
Introduction quality assurance many of the stains that are described in this national standard method nsm are commercially available. Analysis was performed to compare connexin 43 membrane staining in ffpe sections of human lung adenocarcinoma right compared to a negative control without primary antibody left. This staining technique divides bacteria into two groups namely acidfast and non acidfast. Transmission electron microscopy is an effective tool for viewing the morphology and ultrastructure of small biological specimens that are too small to be seen with a traditional light microscope, such as viruses gentile and gelderblom, 2014, kruger et al. Resolution of molecular features is only accomplished at the stain. Gently rinse off the stain with water and shake off the excess. In the present study, although there was a good correlation between the methods, cases in which there was no agreement of results had more midpiece and tail defects falsenegative or a lower value by the sqaiicp system owing to low smi values falsepositive. After haematoxylin staining in the pap stain procedure, og6 is the first counterstain to be used and must be. Pdf modified ultrafast papanicolaou staining technique. Pap smear as the cellular structure is better preserved because the cells are fixed immediately. It is employed in the pap stain to demonstrate keratin. Most commonly used blueing agent is sotts tap water ph 8.
The technique uses a high number of dyes in its procedure. Papanicolaou stain definition of papanicolaou stain by. In addition an example of multicolor staining is available as a separate document entitled an example of staining cells for multicolor flow cytometry. The method for staining in 12x75mm tubes is described below, with a note at the end of the staining protocol describing how the technique is changed when staining in a plate.
Run 95% ethyl alcohol down the slide until the solvent runs clear about 1020. Because of which the stain doesnt penetrate inside the cell and. It usually provides more information about the characteristics of the cell wall thickness. Abnormal findings are often followed up by more sensitive diagnostic procedures and, if.
In the progressive method, the nucleus is stained with hematoxylin to a intensity desired. Negative staining is a simple technique for routine examination of structure. 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. It is a polychromatic stain containing multiple dyes to differentially stain various components of the cells. Negative staining procedure is also called as relief staining. Table of contents immunohistochemical staining methods. Gram staining is a differential staining technique that differentiates bacteria into two groups. Rapid papanicolaou staining method for cervicovaginal.
Papanicolaou stain pap stain protocol ihc world, llc. A method of staining for keratin by application of a modified papanicolaou staining procedure to formalinfixed paraffinembedded tissues is described. Papanicolaou stain also papanicolaou s stain or pap stain is the most important stain utilized in the practice of cytopathology. Alcohols used for wiping the skin may cause hemolysis and artifacts. Immunohistochemical staining methods education guide. The pas staining procedure may also be used for the demonstration of fungal organisms in tissue sections. Preparation of viral samples within biocontainment for. This procedure is extensively used in the diagnosis of tuberculosis and leprosy. Us6284543b1 rapid papanicolaou staining method for. Differential staining differential stains use two or more stains and allow the cells to be categorized into various groups or types. The specimen prestaining fixative solution is a 10%. Residents ppap smearap smear bioline international.
Ihc troubleshooting guide thermo fisher scientific uk. Hematology, histology and cytology stains fisher healthcare protocol hema 3 manual staining system and stat pack for 30second, threestep staining comparable to. The gram stain is a complex and differential staining procedure that remains a useful test performed in microbiology laboratories. Histological methods protocols zfin community wiki. Kansas state veterinary diagnostic laboratory 2,182 views.
Who laboratory manual for the examination and processing. Stain in papanicolaou stain, ea36, ea50 or ea65 of choice for minutes, depending on specimen type and preference of stain intensity. Papanicolaou stain free download as powerpoint presentation. Wholemount staining of biotindextran injected embryos photoconversion of fluorescently labeled profiles for em a simple wholemount staining protocol for bone and or cartilage in adults and larvae.
Gram stain protocols american society of microbiology. The staining solutions are supplied in mmi safestain ampoules, allowing quick handling without the need for pipetting or the preparation. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic staining histological technique developed by george papanikolaou, the father of cytopathology. Acid fast staining procedure place a strip of blotting paper over the slide. Immunohistochemistry of formalinfixed paraffinembedded ffpe cancer tissue. However, in the developing countries, where the cost is a major. Papanicolaou stain is a multichromatic multicolored cytological staining technique developed. Full text of manual histological staining see other formats. Procedure of progressive papanicolaou staining method. A modification of the papanicolaou exfoliative cytology.
Stain in papanicolaou stain, og6 for 30 seconds to 2 minutes, depending on specimen type and preference of stain intensity. Pap staining is used to differentiate cells in smear preparations of various bodily secretions. One such conventional prior art short method which sought to reduce the overall time, was the quick papanicolaou staining procedure for stat specimens developed by the johns hopkins cytopathology laboratory, baltimore, md. Protocol for immunofluorescent staining of mouse frozen.
The papanicolaou test abbreviated as pap test, also known as pap smear, cervical smear, cervical screening, or smear test is a method of cervical screening used to detect potentially precancerous and cancerous processes in the cervix opening of the uterus or womb or colon in both women and men. After haematoxylin staining in the pap stain procedure, og6 is the first counterstain to be used and must be performed before the ea50 or ea65 counterstain as these will prevent satisfactory take up of og6 if they are preformed ahead of it. Get a printable copy pdf file of the complete article 165k, or click on a page image below to browse page by page. Heat gently to steaming and allow to steam for 5 min. Originally, it was introduced to save reagents, lower the staining time, and possibly enhance the quality of staining. Smear should be quickly fixed alcohol gives better nuclear detail than air staining blue green for ribosomes, particularly in parabasal cells and metaplastic squamous cells pink in inactive cells, such as superficial cells. Place the covered slide over a screened water bath and then saturate blotting paper with primary stain ziehls carbol fuschion. In basic staining heat fixing has to be done to be able to make the cells visible whereas with negative staining heat fixing does not have to be done to see the cells more clearly. Protocol for the preparation and fluorescent icc staining. Revised february 2016 papanicolaou stain for cytological. Chapter 32 troubleshooting procedure for artisanlink special staining system 267. Blood samples,fresh and fixed tissue, and cytology slides duration.
Protocol for immunofluorescent staining of mouse frozen sections tissue. Using forceps, tilt each slide to drain off the staining solution. Acidfast stain principle, procedure, interpretation and examples. A read is counted each time someone views a publication summary such as the title, abstract, and list of authors, clicks on a figure, or views or downloads the fulltext.
Once the slides have arrived at the contractors facility, they will be logged in, heat fixed and stained as described. For tap water rinse steps, submerse slide into dish filled with tap water and then place the. Do not allow the staining solution to dry on the slides add fresh solution as required. The bv smears should be packaged so as to protect the glass slides from breaking en route, and maintain ambient temperature during shipping. This stain procedure delineates the acrosome, head, midpiece, and tail of sperm. Comparison of ultrafast papanicolaou stain with the. Modified ultrafast papanicolaou staining technique. Pdf on jan 1, 2012, anil s and others published routine histotechniques, staining and notes on.
Importance of papanicolaou staining for sperm morphologic. The specimen will be covered with a very thin layer of the metal salt, being excluded in areas where an adsorbed macromolecule is present. The procedure is based on the ability of microorganisms to retain color of the stains used during the gram stain reaction. 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. A double staining technique for simultaneous demonstration of astrocytes and microglia in brain sections and astrogial cell cultures june 1991 journal of histochemistry and cytochemistry 395. Consequently, to address the shortcomings of the pap test, a number of derivative methods were developed, such as the ultrafast staining technique, the shortduration papanicolaou stain, the reap. Balachandran iintroductionntroduction cervical cancer is predominantly sexually transmitted and the association between certain oncogenic high risk strains of human papillomavirus hpv and cervical cancer is well documented. Deparaffinize sections by sequential immersion in the following for 2 minutes each. A method for staining a pap test slide specimen for conventional pap test and, particularly a thinprep pap test specimen, in which a number of multiple hydrating, staining and dehydrating step are performed in an equal number of 10 to 15 dippings at about equal dip times of about 1 second. Standard operating procedure bright field immunohistochemistry manual staining sop i. Papanicolaou stain, ea3650 formula, counterstain for use.
Mycobacterium tuberculosis is the most important of this group, as it is responsible for the disease called tuberculosis tb along with some others of this. On the other hand, cells derived from the deeper vaginal layers, which are free from cornification, are stained strongly blue. Gcc diagnostics guarantees that the highest quality reagents are supplied with this product and that the. Achieve rapid results of fine needle aspirations or highrisk specimens with the thermo scientific shandon rapidchrome papanicolaou staining kit.
Ppap smearap smear vvandana mehta, vani vasanth, c. Pdf routine histotechniques, staining and notes on. To ensure quality control and in compliance with the appropriate quality assurance programs, it is necessary to have detailed standard operating procedures sop. Periodic acid schiff pas reagents are for in vitro diagnostic use.