Abstract
We introduce a simple, label-free cytometry technique, based on the spatio-temporal fluctuation analysis of pixel gray levels of a cell image utilizing the Gray Level Information Entropy (GLIE) function. In this study, the difference in GLIE random fluctuations and its biophysical etiology in a comparison cell model of leukemic Jurkat cells and human healthy donor lymphocytes was explored. A combination of common bright field microscopy and a unique imaging dish wherein cells are individually held untethered in a picoliter volume matrix of optical chambers was used. Random GLIE fluctuations were found to be greater in malignant Jurkat cells than in benign lymphocytes, while these fluctuations correlate with intracellular vesicle Mean Square Displacement (MSD) values and are inhibited by myosin-2 and adenosine triphosphate (ATP) inhibitors. These results suggest that the incoherent active forces acting on the cytoskeleton which cause mechanical dissipative fluctuation of the cytoskeletal and related intracellular content are the biophysical cellular mechanism behind the GLIE random fluctuation results. Analysis of the results in Jurkat cells and normal lymphocytes suggests the possible potential of this simple and automated label-free cytometry to identify malignancy, particularly in a diagnostic setup of multiple cell examination.
Similar content being viewed by others
References
Amin HM, Yang Y, Shen Y, Estey EH, Giles FJ, Pierce SA, Kantarjian HM, O’Brien SM, Jilani I, Albitar M (2005) Having a higher blast percentage in circulation than bone marrow: clinical implications in myelodysplastic syndrome and acute lymphoid and myeloid leukemias. Leukemia 19:1567–1572
Arber DA, Borowitz MJ, Cessna M, Etzell J, Foucar K, Hasserjian RP, Rizzo JD, Theil K, Wang SA, Smith AT, Rumble RB, Thomas NE, Vardiman JW (2016) Initial diagnostic workup of acute leukemia. Arch Pathol Lab Med. https://doi.org/10.5858/arpa.2016-0504-CP
Boiteux A, Goldbeter A, Hess B (1975) Control of oscillating glycolysis of yeast by stochastic, periodic, and steady source of substrate: a model and experimental study. Proc Natl Acad Sci USA 72(10):3829–3833
Brangwynne CP, Koenderink GH, MacKintosh FC (2009) Intracellular transport by active diffusion. Trends Cell Biol 19(9):423–427
Corbin EA, Kong F, Lim CT, King WP, Bashir R (2015) Biophysical properties of human breast cancer cells measured using silicon MEMS resonators and atomic force microscopy. Lab Chip 15:839–847
Gatenby RA, Gillies RJ (2004) Why do cancers have high aerobic glycolysis? Nat Rev Cancer 4:891–899
Guck J, Schinkinger S, Lincoln B, Wottawah F, Ebert S, Romeyke M, Lenz D, Erickson HM, Ananthakrishnan R, Mitchell D, Käs J, Ulvick S, Bilby C (2005) Optical deformability as an inherent cell marker for testing malignant transformation and metastatic competence. Biophys J 88(5):3689–3698
Guo M, Ehrlicher AJ, Jensen MH, Renz M, Moore JR, Goldman RD, Lippincott-Schwartz J, Mackintosh FC, Weitz DA (2014) Probing the stochastic, motor-driven properties of the cytoplasm using force spectrum microscopy. Cell 158:822–832
Hudder A, Nathanson L, Deutscher P (2003) Organization of mammalian cytoplasm. Mol Cell Biol 23(24):9318–9326
Hunger SP, Mullighan CG (2015) acute lymphoblastic leukemia in children. N Engl J Med 373:1541–1552
Lau AW, Hoffman BD, Davies A, Crocker JC, Lubensky TC (2003) Microrheology, stress fluctuations and active behavior of living cells. Phys Rev Lett 91(19):198101
Lekka M (2016) Discrimination between normal and cancerous cells using AFM. BioNanoSci 6:65–80
MacKintosh FC, Levine AJ (2008) Non-equilibrium mechanics and dynamics of motor-activated gels. Phys Rev Lett 100:018104
Pacini N, Borziani F (2014) Cancer stem cell theory and the Warburg Effect, Two sides of the same Coin? Int J Mol Sci 15:8893–8930
Poplack DG (1989) Acute lymphoblastic leukemia (Chapter 43). New Directions in Cancer Treatment. Springer, Berlin, pp 546–551
Pui C-H, Evans WE (1998) Acute lymphoblastic leukemia. N Engl J Med 339:605–615
Ravid-Hermesh O, Zurgil N, Shafran Y, Sobolev M, Galmidi M, Badihi Y, Israel LL, Lellouche JP, Lellouche E, Michaeli S, Deutsch M (2014) Real-time quantification of protein expression and translocation at individual cell resolution using imaging-dish-based live cell array. Anal Bioanal Chem 406:7085–7101
Saks V, Beraud N, Wallimann T (2008) Metabolic compartmentation—a system level property of muscle cells. Int J Mol Sci 9(5):751–767
Seyfried TN, Shelton LM (2010) Cancer as a metabolic disease. Nutr Metab 7:7
Sunray M, Zurgil N, Shafran Y, Deutsch M (2002) Determination of individual cell Michaelis-Menten constants. Cytometry 47:8–16
Suresh S (2007) Biomechanics and biophysics of cancer cells. Acta Biomater 3(4):413–438
Wohl I, Zurgil N, Hakuk Y, Sobolev M, Galmidi M, Deutsch M (2015) In situ label-free static cytometry by monitoring spatiotemporal fluctuations of image gray values. J Biomed Opt 20(10):105013
Wohl I, Zurgil N, Hakuk Y, Sobolev M, Deutsch M (2016) In situ evaluation of physiological activity and mitochondrial dysfunction via novo label-free measures based on fluctuation of image gray values. J Anal Bioanal Tech 7:2
Wohl I, Zurgil N, Hakuk Y, Sobolev M, Deutsch M (2017) Fluctuation of information entropy measures in cell image. Entropy 019:565
Yang J-H, Yang L, Qu Z, Weiss JN (2008) Glycolytic oscillations in isolated rabbit ventricular myocytes. J Biol Chem 283(52):36321–36327
Zheng J (2012) Energy metabolism of cancer: Glycolysis versus oxidative phosphorylation (Review). Oncol Lett 4:1151–1157
Acknowledgements
This study was endowed by the Bequest of Moshe-Shimon and Judith Weisbrodt.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflicts of interest
The authors declare no known conflicts of interest, financial or otherwise, associated with this publication and there has been no significant financial support for this work that could have influenced its outcome.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Wohl, I., Zurgil, N., Hakuk, Y. et al. Discrimination of leukemic Jurkat cells from normal lymphocytes via novo label-free cytometry based on fluctuation of image gray values. Eur Biophys J 48, 267–275 (2019). https://doi.org/10.1007/s00249-019-01351-w
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00249-019-01351-w