Flow cytometry and laser scanning cytometry, a comparison of techniques
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Flow and laser scanning cytometry are used extensively in research and clinical settings. These techniques provide clinicians and scientists information about cell functioning in a variety of health and disease states. An in-depth knowledge and understanding of cytometry techniques can enhance interpretation of current research findings. Our goal with this review is to reacquaint clinicians and scientists with information concerning differences between flow and laser scanning cytometry by comparing their capabilities and applications.
A Pubmed abstract search was conducted for articles on research, reviews and current texts relating to origins and use of flow and laser scanning cytometry. Attention was given to studies describing application of these techniques in the clinical setting.
Both techniques exploit interactions between the physical properties of light. Data are immediately and automatically acquired; they are distinctly different. Flow cytometry provides valuable rapid information about a wide variety of cellular or particle characteristics. This technique does not provide the scanned high resolution image analysis needed for investigators to localize areas of interest within the cell for quantification. Flow cytometry requires that the sample contain a large amount disaggregated, single, suspended cells. Laser scanning cytometry is slide-based and does not require as large of a sample. The tissue sample is affixed to a slide allowing repeated sample analyses. These cytometry techniques are used in the clinical setting to understand pathophysiological derangements associated with many diseases; cardiovascular disease, diabetes, acute lung injury, hemorrhagic shock, surgery, cancer and Alzheimer’s disease.
Understanding the dif- ferences between FCM and LSCM can assist investigators in planning and design of their research or clinical testing. Researchers and clinicians optimize these technique capa- bilities with the cellular characteristics they wish to measure delineating molecular and cellular events occurring in health and disease. Discovery of mechanisms in cells using FCM and LSCM provide evidence needed to guide future treatment and interventions.
- Tarnok A, Gerstner AO. Clinical applications of laser scanning cytometry. Cytometry. 2002;50(3):133–43. CrossRef
- Pozarowski P, Holden E, Darzynkiewicz Z. Laser scanning cytometry: principles and applications. Methods Mol Biol. 2006;319:165–92. CrossRef
- Valet G. Past and present concepts in flow cytometry: a European perspective. J Biol Regul Homeost Agents. 2003;17(3):213–22.
- Darzynkiewicz Z, Crissman H, Jacobberger JW. Cytometry of the cell cycle: cycling through history. Cytometry A. 2004;58(1):21–32. CrossRef
- Kamentsky LA. Laser scanning cytometry. Methods Cell Biol. 2001;63:51–87. CrossRef
- Nolan JP, Yang L. The flow of cytometry into systems biology. Brief Funct Genomic Proteomic. 2007;6(2):81–90. CrossRef
- Shapiro H. Practical flow cytometry. 4th ed. Hoboken: Willey-Liss; 2003.
- Rieseberg M, Kasper C, Reardon KF, Scheper T. Flow cytometry in biotechnology. Appl Microbiol Biotechnol. 2001;56(3–4):350–60. CrossRef
- Cram LS. Flow cytometry, an overview. Methods Cell Sci. 2002;24(1–3):1–9. CrossRef
- Luther E, Kamentsky L, Henriksen M, Holden E. Next-generation laser scanning cytometry. Methods Cell Biol. 2004;75:185–218. CrossRef
- Kamentsky LA, Burger DE, Gershman RJ, Kamentsky LD, Luther E. Slide-based laser scanning cytometry. Acta Cytol. 1997;41(1):123–43.
- Kamentsky LA, Kamentsky LD, Fletcher JA, Kurose A, Sasaki K. Methods for automatic multiparameter analysis of fluorescence in situ hybridized specimens with a laser scanning cytometer. Cytometry. 1997;27(2):117–25. CrossRef
- Darzynkiewicz Z, Bedner E, Li X, Gorczyca W, Melamed MR. Laser-scanning cytometry: a new instrumentation with many applications. Exp Cell Res. 1999;249(1):1–12. CrossRef
- Wijsman JA, Obert LA, Paulissen J, Garrido R, Toy KA, Dunstan RW. A practical method to determine the amount of tissue to analyze using laser scanning cytometry. Cytometry A. 2007;71(7):501–8.
- Cram LS, Arndt-Jovin D. Mack Jett Fulwyler, pioneer of flow cytometry and flow sorting (1936–2001). Cytometry A. 2005;67(2):53–4. cover.
- Mittag A, Lenz D, Gerstner AO, Tarnok A. Hyperchromatic cytometry principles for cytomics using slide based cytometry. Cytometry A. 2006;69(7):691–703.
- Oswald J, Jorgensen B, Pompe T, et al. Comparison of flow cytometry and laser scanning cytometry for the analysis of CD34+ hematopoietic stem cells. Cytometry A. 2004;57(2):100–7. CrossRef
- Bedner E, Burfeind P, Gorczyca W, Melamed MR, Darzynkiewicz Z. Laser scanning cytometry distinguishes lymphocytes, monocytes, and granulocytes by differences in their chromatin structure. Cytometry. 1997;29(3):191–6. CrossRef
- Holden JA. DNA topoisomerases as anticancer drug targets: from the laboratory to the clinic. Curr Med Chem Anticancer Agents. 2001;1(1):1–25. CrossRef
- Pozarowski P, Darzynkiewicz Z. Analysis of cell cycle by flow cytometry. Methods Mol Biol. 2004;281:301–11.
- Vermes I, Haanen C, Reutelingsperger C. Flow cytometry of apoptotic cell death. J Immunol Methods. 2000;243(1–2):167–90. CrossRef
- Bedner E, Li X, Gorczyca W, Melamed MR, Darzynkiewicz Z. Analysis of apoptosis by laser scanning cytometry. Cytometry. 1999;35(3):181–95. CrossRef
- Darzynkiewicz Z, Bedner E, Traganos F, Murakami T. Critical aspects in the analysis of apoptosis and necrosis. Hum Cell. 1998;11(1):3–12.
- Darzynkiewicz Z, Juan G, Li X, Gorczyca W, Murakami T, Traganos F. Cytometry in cell necrobiology: analysis of apoptosis and accidental cell death (necrosis). Cytometry. 1997;27(1):1–20. CrossRef
- Matyus AE. Flow cytometry and cell sorting (Vol. 1: Techniques). New York: Plenum Press; 1991.
- DeRosa S, Benchley J, Roederer M. Beyond six colors: a new era in flow cytometry. Nat Med. 2003;9(1):112–7. CrossRef
- Deptala A, Bedner E, Darzynkiewicz Z. Unique analytical capabilities of laser scanning cytometry (LSC) that complement flow cytometry. Folia Histochem Cytobiol. 2001;39(2):87–9.
- Herzenberg LA, Tung J, Moore WA, Parks DR. Interpreting flow cytometry data: a guide for the perplexed. Nat Immunol. 2006;7(7):681–5. CrossRef
- Roederer M, Darzynkiewicz Z, Parks DR. Guidelines for the presentation of flow cytometric data. Methods Cell Biol. 2004;75:241–56. CrossRef
- Wood JC, Hoffman RA. Evaluating fluorescence sensitivity on flow cytometers: an overview. Cytometry. 1998;33(2):256–9. CrossRef
- Brown M, Wittwer C. Flow cytometry: principles and clinical applications in hematology. Clin Chem. 2000;46(8 Pt 2):1221–9.
- Bigalke B, Stellos K, Stakos D, et al. Influence of platelet count on the expression of platelet collagen receptor glycoprotein VI (GPVI) in patients with acute coronary syndrome. Thromb Haemost. 2009;101(5):911–5.
- Fox SC, May JA, Shah A, Neubert U, Heptinstall S. Measurement of platelet P-selectin for remote testing of platelet function during treatment with clopidogrel and/or aspirin. Platelets. 2009;20(4):250–9. CrossRef
- Sibal, L., Aldibbiat, A., Agarwal, S.C., et al. Circulating endothelial progenitor cells, endothelial function, carotid intima-media thickness and circulating markers of endothelial dysfunction in people with type 1 diabetes without macrovascular disease or microalbuminuria. Diabetologia 2009; 52(8): 1464–1473
- Taatjes DJ, Wadsworth MP, Quinn AS, Rand JH, Bovill EG, Sobel BE. Imaging aspects of cardiovascular disease at the cell and molecular level. Histochem Cell Biol. 2008;130(2):235–45. CrossRef
- Schneider DJ, Hayes M, Wadsworth M, et al. Attenuation of neointimal vascular smooth muscle cellularity in atheroma by plasminogen activator inhibitor type 1 (PAI-1). J Histochem Cytochem. 2004;52(8):1091–9. CrossRef
- Kolackova M, Kudlova M, Kunes P, et al. Early expression of FcgammaRI (CD64) on monocytes of cardiac surgical patients and higher density of monocyte anti-inflammatory scavenger CD163 receptor in “on-pump” patients. Mediators Inflamm 2008;235461.
- Ritter L, Funk J, Schenkel L, et al. Inflammatory and hemodynamic changes in the cerebral microcirculation of aged rats after global cerebral ischemia and reperfusion. Microcirculation. 2008;15(4):297–310. CrossRef
- Vega D, Badami CD, Caputo FJ, et al. The influence of the type of resuscitation fluid on gut injury and distant organ injury in a rat model of trauma/hemorrhagic shock. J Trauma. 2008;65(2):409–14. discussion 414–405. CrossRef
- Jones R, Capen DE, Jacobson M, Cohen KS, Scadden DT, Duda DG. VEGFR2PDGFRbeta circulating precursor cells participate in capillary restoration after hyperoxia acute lung injury (HALI). J Cell Mol Med 2009; 13(9B), 3720–3729.
- Suzuki Y, Nishio K, Takeshita K, et al. Effect of steroid on hyperoxia-induced ICAM-1 expression in pulmonary endothelial cells. Am J Physiol Lung Cell Mol Physiol. 2000;278(2):L245–52.
- Chang LT, Sun CK, Chiang CH, Wu CJ, Chua S, Yip HK. Impact of simvastatin and losartan on antiinflammatory effect: in vitro study. J Cardiovasc Pharmacol. 2007;49(1):20–6. CrossRef
- Buder-Hoffmann S, Palmer C, Vacek P, Taatjes D, Mossman B. Different accumulation of activated extracellular signal-regulated kinases (ERK 1/2) and role in cell-cycle alterations by epidermal growth factor, hydrogen peroxide, or asbestos in pulmonary epithelial cells. Am J Respir Cell Mol Biol. 2001;24(4):405–13.
- Jin D, Port J, Korst R. Lee P, Li Z, McDonald K, Ferrara C, Meherally D, Rafii S, Altorki N. Surgical resection normalizes the hemangiogenic profile in patients with non-small cell lung cancer. Paper presented at the American Association for Thoracic Surgery. 2007.
- Ishikawa M, Nishioka M, Hanaki N, et al. Perioperative immune responses in cancer patients undergoing digestive surgeries. World J Surg Oncol. 2009;7:7. CrossRef
- Park MS, Lee HM, Hahn SB, et al. The association of the activation-inducible tumor necrosis factor receptor and ligand with lumbar disc herniation. Yonsei Med J. 2007;48(5):839–46. CrossRef
- Henry M, Davidson L, Cohen Z, McDonagh PF, Nolan PE, Ritter LS. Whole blood aggregation, coagulation, and markers of platelet activation in diet-induced diabetic C57BL/6J mice. Diabetes Res Clin Pract. 2009;84(1):11–8. CrossRef
- Chen JZ, Zhang FR, Tao QM, Wang XX, Zhu JH. Number and activity of endothelial progenitor cells from peripheral blood in patients with hypercholesterolaemia. Clin Sci (Lond). 2004;107(3):273–80. CrossRef
- Turina M, Miller FN, Tucker CF, Polk HC. Short-term hyperglycemia in surgical patients and a study of related cellular mechanisms. Ann Surg. 2006;243(6):845–51. discussion 851–843. CrossRef
- Yow CM, Chen JY, Mak NK, Cheung NH, Leung AW. Cellular uptake, subcellular localization and photodamaging effect of temoporfin (mTHPC) in nasopharyngeal carcinoma cells: comparison with hematoporphyrin derivative. Cancer Lett. 2000;157(2):123–31. CrossRef
- Paiva B, Vidriales MB, Cervero J, et al. Multiparameter flow cytometric remission is the most relevant prognostic factor for multiple myeloma patients who undergo autologous stem cell transplantation. Blood. 2008;112(10):4017–23. CrossRef
- Al-Mawali A, Gillis D, Lewis I. The role of multiparameter flow cytometry for detection of minimal residual disease in acute myeloid leukemia. Am J Clin Pathol. 2009;131(1):16–26. CrossRef
- Landgren O, Albitar M, Ma W, et al. B-cell clones as early markers for chronic lymphocytic leukemia. N Engl J Med. 2009;360(7):659–67. CrossRef
- Chen H, Sun B, Pan S, Jiang H, Sun X. Dihydroartemisinin inhibits growth of pancreatic cancer cells in vitro and in vivo. Anticancer Drugs. 2009;20(2):131–40. CrossRef
- Lu YY, Chen TS, Qu JL, Pan WL, Sun L, Wei XB. Dihydroartemisinin (DHA) induces caspase-3-dependent apoptosis in human lung adenocarcinoma ASTC-a-1 cells. J Biomed Sci. 2009;16:16. CrossRef
- Ueno C, Fukatsu K, Kang W, et al. Route and type of nutrition influence nuclear factor kappaB activation in peritoneal resident cells. Shock. 2005;24(4):382–7. CrossRef
- Tomita K, Chikumi H, Tokuyasu H, et al. Functional assay of NF-kappaB translocation into nuclei by laser scanning cytometry: inhibitory effect by dexamethasone or theophylline. Naunyn Schmiedebergs Arch Pharmacol. 1999;359(4):249–55. CrossRef
- Zuba-Surma EK, Kucia M, Abdel-Latif A, Lillard JW Jr, Ratajczak MZ. The ImageStream System: a key step to a new era in imaging. Folia Histochem Cytobiol. 2007;45(4):279–90.
- Flow cytometry and laser scanning cytometry, a comparison of techniques
Journal of Clinical Monitoring and Computing
Volume 24, Issue 4 , pp 251-259
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- Springer Netherlands
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- flow cytometry
- laser scanning cytometry
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- Author Affiliations
- 1. School of Nursing, University of Kansas, Mail Stop 4043, 3901 Rainbow Blvd., Kansas City, KS, 66160, USA
- 2. Department of Molecular Biosciences, University of Kansas, Lawrence, KS, 66045, USA
- 3. Flow Cytometry Laboratory, University of Kansas, 3901 Rainbow Blvd., Kansas City, KS, 66160, USA