Cytocentrifuge

Last updated May 10, 2024

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. ^[11] 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".^[12] 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

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<span class="mw-page-title-main">Georgios Papanikolaou</span> Greek pathologist (1883–1962)

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".

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 is 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.

Poikilocytosis is variation in the shapes of red blood cells. Poikilocytes may be oval, teardrop-shaped, sickle-shaped or irregularly contracted.

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.

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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.

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References

↑ McManus L, Mitchell R (1 August 2014). Pathobiology of Human Disease: A Dynamic Encyclopedia of Disease Mechanisms. Elsevier Science. p. 3365. ISBN 978-0-12-386457-4.
↑ Turgeon ML (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 BO (2004). "Principles of Cytocentrifugation". Laboratory Medicine. 35 (7): 434–437. doi: 10.1309/FTT59GWKDWH69FB0 . ISSN 0007-5027.
1 2 3 Brunzel NA (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.
↑ Galagan KA (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.
↑ Shambayati B (17 February 2011). Cytopathology. OUP Oxford. p. 24. ISBN 978-0-19-953392-3.
↑ Keohane EM, Smith L, Walenga JM (19 February 2015). Rodak's Hematology: Clinical Principles and Applications. Elsevier Health Sciences. pp. 270–1. ISBN 978-0-323-32716-9.
1 2 Gill G (19 October 2012). "Chapter 6: Cytocentrifugation". Cytopreparation: Principles & Practice. Springer Science & Business Media. pp. 73–84. ISBN 978-1-4614-4932-4.
↑ Mahon CR, Lehman DC, Manuselis G (25 March 2014). Textbook of Diagnostic Microbiology - E-Book. Elsevier Health Sciences. p. 129. ISBN 978-0-323-29262-7.
1 2 Harmening D (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.
↑ Vembadi A, Menachery A, Qasaimeh MA (2019). "Cell Cytometry: Review and Perspective on Biotechnological Advances". Frontiers in Bioengineering and Biotechnology. 7: 147. doi: 10.3389/fbioe.2019.00147 . PMC 6591278 . PMID 31275933.
↑ Watson P (September 1966). "A slide centrifuge: an apparatus for concentrating cells in suspension onto a microscope slide". The Journal of Laboratory and Clinical Medicine. 68 (3): 494–501. PMID 5922759.

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[McManus2014-1] McManus L, Mitchell R (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] Turgeon ML (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 BO (2004). "Principles of Cytocentrifugation". Laboratory Medicine. 35 (7): 434–437. doi: 10.1309/FTT59GWKDWH69FB0 . ISSN 0007-5027.

[Brunzel2016-4] 1 2 3 Brunzel NA (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] Galagan KA (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] Shambayati B (17 February 2011). Cytopathology. OUP Oxford. p. 24. ISBN 978-0-19-953392-3.

[Keohane2015-7] Keohane EM, Smith L, Walenga JM (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 Gill G (19 October 2012). "Chapter 6: Cytocentrifugation". Cytopreparation: Principles & Practice. Springer Science & Business Media. pp. 73–84. ISBN 978-1-4614-4932-4.

[MahonLehman2014-9] Mahon CR, Lehman DC, Manuselis G (25 March 2014). Textbook of Diagnostic Microbiology - E-Book. Elsevier Health Sciences. p. 129. ISBN 978-0-323-29262-7.

[Harmening2009-10] 1 2 Harmening D (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.

[11] Vembadi A, Menachery A, Qasaimeh MA (2019). "Cell Cytometry: Review and Perspective on Biotechnological Advances". Frontiers in Bioengineering and Biotechnology. 7: 147. doi: 10.3389/fbioe.2019.00147 . PMC 6591278 . PMID 31275933.

[Watson1966-12] Watson P (September 1966). "A slide centrifuge: an apparatus for concentrating cells in suspension onto a microscope slide". The Journal of Laboratory and Clinical Medicine. 68 (3): 494–501. PMID 5922759.

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