Cytocentrifuge

Last updated August 29, 2023

A cytocentrifuge, sometimes referred to as a cytospin,^[1] is a specialized centrifuge used to concentrate cells in fluid specimens onto a microscope slide so that they can be stained and examined.^[2] Cytocentrifuges are used in various areas of the clinical laboratory, such as cytopathology, hematology and microbiology, as well as in biological research. The method can be used on many different types of specimens, including fine needle aspirates, cerebrospinal fluid, serous and synovial fluid, and urine.^[3]

Procedure

To prepare cytocentrifuge smears, a funnel assembly is attached to the front of a microscope slide. The surface of the funnel assembly that is in contact with the slide is lined with filter paper to absorb excess fluid. A few drops of fluid are placed in the funnel. The assembly is placed in the cytocentrifuge, which operates at a low force (600–800 x g) to preserve cellular structure.^[4] Centrifugal force pushes the fluid through the funnel's opening and concentrates the cells in a small area of the slide. The centrifugation process concentrates cells by about twenty-fold^[5] and creates a one-cell-thick monolayer, allowing for assessment of cellular morphology. The slide can then be fixed and stained.^[6]^[7]

Applications

Some applications of cytocentrifuges include:

Performing differential cell counts on body fluids, such as serous, synovial and cerebrospinal fluid ^[4]
Cytopathology examination of liquid specimens such as body fluids and fine needle aspirates ^[3]^[8]
Gram staining of fluid specimens for identification of microorganisms ^[9]

Limitations

The cytocentrifugation process can cause cells to appear distorted. Cells located at the centre of the smear may look compressed compared to cells at the periphery. Cell nuclei may develop artifactual clefts, lobes, or holes,^[4] and the cytoplasm may appear vacuolated or develop irregular projections. Cytoplasmic granules may be pushed to the periphery of the cell. If the cell count is high, cells may be distorted due to crowding; therefore, samples with high cell counts are diluted prior to smear preparation.^[10]

History

Examination of cells in body fluids was historically performed using a hemocytometer, a chamber designed for counting cells microscopically. This technique was limited by poor discrimination between cell types (cells could only be classified as mononuclear or polymorphonuclear) and the low number of cells present in unconcentrated body fluids. Moreover, this technique did not produce a permanent record of the specimen.^[10] In a 1966 paper, Watson P. described the first cytocentrifuge, calling it "an apparatus for concentrating cells in suspension onto a microscope slide".^[11] The device was sold commercially in the 1970s and in 1983 it was patented by Shandon (now Thermo Scientific). As of 2012, numerous brands of cytocentrifuge exist on the market.^[8]

Related Research Articles

Pathology is the study of the causes and effects of disease or injury. The word pathology also refers to the study of disease in general, incorporating a wide range of biology research fields and medical practices. However, when used in the context of modern medical treatment, the term is often used in a narrower fashion to refer to processes and tests that fall within the contemporary medical field of "general pathology", an area which includes a number of distinct but inter-related medical specialties that diagnose disease, mostly through analysis of tissue and human cell samples. Idiomatically, "a pathology" may also refer to the predicted or actual progression of particular diseases, and the affix pathy is sometimes used to indicate a state of disease in cases of both physical ailment and psychological conditions. A physician practicing pathology is called a pathologist.

Anatomical pathology (Commonwealth) or Anatomic pathology (U.S.) is a medical specialty that is concerned with the diagnosis of disease based on the macroscopic, microscopic, biochemical, immunologic and molecular examination of organs and tissues. Over the 20th century, surgical pathology has evolved tremendously: from historical examination of whole bodies (autopsy) to a more modernized practice, centered on the diagnosis and prognosis of cancer to guide treatment decision-making in oncology. Its modern founder was the Italian scientist Giovan Battista Morgagni from Forlì.

Anemia or anaemia is a blood disorder in which the blood has a reduced ability to carry oxygen due to a lower than normal number of red blood cells, or a reduction in the amount of hemoglobin. The name is derived from Ancient Greek: ἀναιμία anaimia, meaning 'lack of blood', from ἀν- an-, 'not' and αἷμα haima, 'blood'. When anemia comes on slowly, the symptoms are often vague, such as tiredness, weakness, shortness of breath, headaches, and a reduced ability to exercise. When anemia is acute, symptoms may include confusion, feeling like one is going to pass out, loss of consciousness, and increased thirst. Anemia must be significant before a person becomes noticeably pale. Symptoms of anemia depend on how quickly hemoglobin decreases. Additional symptoms may occur depending on the underlying cause. Preoperative anemia can increase the risk of needing a blood transfusion following surgery. Anemia can be temporary or long term and can range from mild to severe.

<span class="mw-page-title-main">Georgios Papanikolaou</span> Greek pathologist

Georgios Nikolaou Papanikolaou was a Greek physician, zoologist and microscopist who was a pioneer in cytopathology and early cancer detection, and inventor of the "Pap smear".

Cytopathology is a branch of pathology that studies and diagnoses diseases on the cellular level. The discipline was founded by George Nicolas Papanicolaou in 1928. Cytopathology is generally used on samples of free cells or tissue fragments, in contrast to histopathology, which studies whole tissues. Cytopathology is frequently, less precisely, called "cytology", which means "the study of cells".

Romanowsky staining, also known as Romanowsky–Giemsa staining, is a prototypical staining technique that was the forerunner of several distinct but similar stains widely used in hematology and cytopathology. Romanowsky-type stains are used to differentiate cells for microscopic examination in pathological specimens, especially blood and bone marrow films, and to detect parasites such as malaria within the blood. Stains that are related to or derived from the Romanowsky-type stains include Giemsa, Jenner, Wright, Field, May–Grünwald and Leishman stains. The staining technique is named after the Russian physician Dmitri Leonidovich Romanowsky (1861–1921), who was one of the first to recognize its potential for use as a blood stain.

A complete blood count (CBC), also known as a full blood count (FBC), is a set of medical laboratory tests that provide information about the cells in a person's blood. The CBC indicates the counts of white blood cells, red blood cells and platelets, the concentration of hemoglobin, and the hematocrit. The red blood cell indices, which indicate the average size and hemoglobin content of red blood cells, are also reported, and a white blood cell differential, which counts the different types of white blood cells, may be included.

The pericardium, also called pericardial sac, is a double-walled sac containing the heart and the roots of the great vessels. It has two layers, an outer layer made of strong inelastic connective tissue, and an inner layer made of serous membrane. It encloses the pericardial cavity, which contains pericardial fluid, and defines the middle mediastinum. It separates the heart from interference of other structures, protects it against infection and blunt trauma, and lubricates the heart's movements.

Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology, in cytology, and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses of diseases at the microscopic level. Stains may be used to define biological tissues, cell populations, or organelles within individual cells.

A blood smear, peripheral blood smear or blood film is a thin layer of blood smeared on a glass microscope slide and then stained in such a way as to allow the various blood cells to be examined microscopically. Blood smears are examined in the investigation of hematological (blood) disorders and are routinely employed to look for blood parasites, such as those of malaria and filariasis.

Histopathology refers to the microscopic examination of tissue in order to study the manifestations of disease. Specifically, in clinical medicine, histopathology refers to the examination of a biopsy or surgical specimen by a pathologist, after the specimen has been processed and histological sections have been placed onto glass slides. In contrast, cytopathology examines free cells or tissue micro-fragments.

Papanicolaou stain is a multichromatic (multicolored) cytological staining technique developed by George Papanicolaou in 1942. The Papanicolaou stain is one of the most widely used stains in cytology, where it is used to aid pathologists in making a diagnosis. Although most notable for its use in the detection of cervical cancer in the Pap test or Pap smear, it is also used to stain non-gynecological specimen preparations from a variety of bodily secretions and from small needle biopsies of organs and tissues. Papanicolaou published three formulations of this stain in 1942, 1954, and 1960.

In physiology, serous fluid or serosal fluid is any of various body fluids resembling serum, that are typically pale yellow or transparent and of a benign nature. The fluid fills the inside of body cavities. Serous fluid originates from serous glands, with secretions enriched with proteins and water. Serous fluid may also originate from mixed glands, which contain both mucous and serous cells. A common trait of serous fluids is their role in assisting digestion, excretion, and respiration.

Diff-Quik is a commercial Romanowsky stain variant used to rapidly stain and differentiate a variety of pathology specimens. It is most frequently used for blood films and cytopathological smears, including fine needle aspirates. The Diff-Quik procedure is based on a modification of the Wright-Giemsa stain pioneered by Harleco in the 1970s, and has advantages over the routine Wright-Giemsa staining technique in that it reduces the 4-minute process into a much shorter operation and allows for selective increased eosinophilic or basophilic staining depending upon the time the smear is left in the staining solutions.

<span class="mw-page-title-main">Medical laboratory</span> Principles of management with special reference to medical science

A medical laboratory or clinical laboratory is a laboratory where tests are conducted out on clinical specimens to obtain information about the health of a patient to aid in diagnosis, treatment, and prevention of disease. Clinical medical laboratories are an example of applied science, as opposed to research laboratories that focus on basic science, such as found in some academic institutions.

Neutrophil hypersegmentation can be defined as the presence of neutrophils whose nuclei have six or more lobes or the presence of more than 3% of neutrophils with at least five nuclear lobes. This is a clinical laboratory finding. It is visualized by drawing blood from a patient and viewing the blood smeared on a slide under a microscope. Normal neutrophils are uniform in size, with an apparent diameter of about 13 μm in a film. When stained, neutrophils have a segmented nucleus and pink/orange cytoplasm under light microscope. The majority of neutrophils have three nuclear segments (lobes) connected by tapering chromatin strands. A small percentage have four lobes, and occasionally five lobes may be seen. Up to 8% of circulating neutrophils are unsegmented.

Liquid-based cytology is a method of preparing samples for examination in cytopathology. The sample is collected, normally by a small brush, in the same way as for a conventional smear test, but rather than the smear being transferred directly to a microscope slide, the sample is deposited into a small bottle of preservative liquid. At the laboratory, the liquid is treated to remove other elements such as mucus before a layer of cells is placed on a slide.

Toxic vacuolation, also known as toxic vacuolization, is the formation of vacuoles in the cytoplasm of neutrophils in response to severe infections or inflammatory conditions.

A white blood cell differential is a medical laboratory test that provides information about the types and amounts of white blood cells in a person's blood. The test, which is usually ordered as part of a complete blood count (CBC), measures the amounts of the five normal white blood cell types – neutrophils, lymphocytes, monocytes, eosinophils and basophils – as well as abnormal cell types if they are present. These results are reported as percentages and absolute values, and compared against reference ranges to determine whether the values are normal, low, or high. Changes in the amounts of white blood cells can aid in the diagnosis of many health conditions, including viral, bacterial, and parasitic infections and blood disorders such as leukemia.

Alder–Reilly anomaly, or Alder anomaly, is an inherited abnormality of white blood cells associated with mucopolysaccharidosis. When blood smears and bone marrow preparations from patients with Alder–Reilly anomaly are stained and examined microscopically, large, coarse granules may be seen in their neutrophils, monocytes, and lymphocytes. The condition may be mistaken for toxic granulation, a type of abnormal granulation in neutrophils that occurs transiently in inflammatory conditions.

References

↑ Linda McManus; Richard Mitchell (1 August 2014). Pathobiology of Human Disease: A Dynamic Encyclopedia of Disease Mechanisms. Elsevier Science. p. 3365. ISBN 978-0-12-386457-4.
↑ Mary Louise Turgeon (23 March 2015). Linné & Ringsrud's Clinical Laboratory Science: Concepts, Procedures, and Clinical Applications (7th ed.). Elsevier Mosby. p. 146. ISBN 978-0-323-22545-8.
1 2 Stokes, Barry O. (2004). "Principles of Cytocentrifugation". Laboratory Medicine. 35 (7): 434–437. doi: 10.1309/FTT59GWKDWH69FB0 . ISSN 0007-5027.
1 2 3 Nancy A. Brunzel (5 November 2016). "Chapter 17: Body fluid analysis". Fundamentals of Urine and Body Fluid Analysis. Elsevier Health Sciences. pp. 356–58. ISBN 978-0-323-39636-3.
↑ Katherine A. Galagan (2006). Color Atlas of Body Fluids: An Illustrated Field Guide Based on Proficiency Testing. College of American Pathologists. pp. 13–14. ISBN 978-0-930304-91-1.
↑ Behdad Shambayati (17 February 2011). Cytopathology. OUP Oxford. p. 24. ISBN 978-0-19-953392-3.
↑ Elaine M. Keohane; Larry Smith; Jeanine M. Walenga (19 February 2015). Rodak's Hematology: Clinical Principles and Applications. Elsevier Health Sciences. pp. 270–1. ISBN 978-0-323-32716-9.
1 2 Gary Gill (19 October 2012). "Chapter 6: Cytocentrifugation". Cytopreparation: Principles & Practice. Springer Science & Business Media. pp. 73–84. ISBN 978-1-4614-4932-4.
↑ Connie R. Mahon; Donald C. Lehman; George Manuselis (25 March 2014). Textbook of Diagnostic Microbiology - E-Book. Elsevier Health Sciences. p. 129. ISBN 978-0-323-29262-7.
1 2 Denise Harmening (2009). "Chapter 30: Body fluid examination: qualitative, quantitative and morphologic analysis". Clinical Hematology and Fundamentals of Hemostasis (5th ed.). F. A. Davis Company. pp. 720–757. ISBN 978-0-8036-1732-2.
↑ Watson P (1966). "A slide centrifuge: an apparatus for concentrating cells in suspension onto a microscope slide". J Lab Clin Med. 68 (3): 494–501. PMID 5922759.

This page is based on this Wikipedia article
Text is available under the CC BY-SA 4.0 license; additional terms may apply.
Images, videos and audio are available under their respective licenses.

[McManus2014-1] Linda McManus; Richard Mitchell (1 August 2014). Pathobiology of Human Disease: A Dynamic Encyclopedia of Disease Mechanisms. Elsevier Science. p. 3365. ISBN 978-0-12-386457-4.

[Turgeon2015-2] Mary Louise Turgeon (23 March 2015). Linné & Ringsrud's Clinical Laboratory Science: Concepts, Procedures, and Clinical Applications (7th ed.). Elsevier Mosby. p. 146. ISBN 978-0-323-22545-8.

[Stokes2004-3] 1 2 Stokes, Barry O. (2004). "Principles of Cytocentrifugation". Laboratory Medicine. 35 (7): 434–437. doi: 10.1309/FTT59GWKDWH69FB0 . ISSN 0007-5027.

[Brunzel2016-4] 1 2 3 Nancy A. Brunzel (5 November 2016). "Chapter 17: Body fluid analysis". Fundamentals of Urine and Body Fluid Analysis. Elsevier Health Sciences. pp. 356–58. ISBN 978-0-323-39636-3.

[CAP2006-5] Katherine A. Galagan (2006). Color Atlas of Body Fluids: An Illustrated Field Guide Based on Proficiency Testing. College of American Pathologists. pp. 13–14. ISBN 978-0-930304-91-1.

[Shambayati2011-6] Behdad Shambayati (17 February 2011). Cytopathology. OUP Oxford. p. 24. ISBN 978-0-19-953392-3.

[Keohane2015-7] Elaine M. Keohane; Larry Smith; Jeanine M. Walenga (19 February 2015). Rodak's Hematology: Clinical Principles and Applications. Elsevier Health Sciences. pp. 270–1. ISBN 978-0-323-32716-9.

[Gill2012-8] 1 2 Gary Gill (19 October 2012). "Chapter 6: Cytocentrifugation". Cytopreparation: Principles & Practice. Springer Science & Business Media. pp. 73–84. ISBN 978-1-4614-4932-4.

[MahonLehman2014-9] Connie R. Mahon; Donald C. Lehman; George Manuselis (25 March 2014). Textbook of Diagnostic Microbiology - E-Book. Elsevier Health Sciences. p. 129. ISBN 978-0-323-29262-7.

[Harmening2009-10] 1 2 Denise Harmening (2009). "Chapter 30: Body fluid examination: qualitative, quantitative and morphologic analysis". Clinical Hematology and Fundamentals of Hemostasis (5th ed.). F. A. Davis Company. pp. 720–757. ISBN 978-0-8036-1732-2.

[Watson1966-11] Watson P (1966). "A slide centrifuge: an apparatus for concentrating cells in suspension onto a microscope slide". J Lab Clin Med. 68 (3): 494–501. PMID 5922759.

[1]

[2]

[3]

[4]

[5]

[6]

[7]

[8]

[9]

[10]

[11]