**Author details**

Hisham Mohamed *Egypt Nanotechnology Center (EGNC), Egypt* 

## **6. References**

	- [12] Chou C, Austin RH, Bakajin O, Tegenfeldt JO, Castelino JA, Chan SS, Cox EC, Craighead, H, Darnton N, Duke T, Han J, Turner S. Sorting biomolecules with microdevices.. Electrophoresis 2000: (*21)* 81-90.

Use of Microfluidic Technology for Cell Separation 223

[30] Spizzo G, Gastl G, Wolf D, Gunsilius E, Steurer M, Fong D, Amberger A, Margreiter R, Obrist P. Correlation of COX-2 and Ep-CAM overexpression in human invasive breast

[31] Spizzo G, Went P, Dirnhofer S, Obrist P, Simon R, Spichtin H, Maurer R, Metzger U, Castelberg B, Bart R, Stopatschinskaya S, Kochli O, Haas P, Mross F, Zuber M, Dietrich H, Bischoff S, Mirlacher M, Sauter G, Gastl G. High Ep-CAM expression is associated with poor prognosis in node positive breast cancer. Breast Cancer Research and

[32] Talasaz AH, Powell AA, Huber DE. Berbee JG, Roh KH, Yu W, Xiao W, Davis MM, Pease RF, Mindrinos MN, Jeffrey SS, Davic RW. Isolating highly enriched populations of circulating epithelial cells and other rare cells from blood using a magnetic sweeper

[33] Kovacs GTA. Micromachined Transducers Sourcebook. Boston: McGraw Hill; 1998 [34] Madou MJ. Fundamentals of Microfabrication 2nd edn. Boca Raton: CRC Press: 2002.

[40] Sin A, Murthy SK, Revzin A, Tompkins RG, Toner M. Enrichment Using Antibody-Coated Microfluidic Chambers in Shear Flow: Model Mixtures of Human Lymphocytes.

[41] Fu AY, Chou HP, Spence C, Arnold FH, Quake SR. An Integrated Microfabricated Cell

[42] Revzin A, Sekine K, Sin A, Tompkins RG, Toner M. Development of a microfabricated cytometry platform for characterization and sorting of individual leukocytes. Lab on a

[43] Chang WC, Lee LP, Liepmann D. Biomimetic technique for adhesion-based collection and separation of cells in a microfluidic channel. Lab on a chip 2005: (5) 64-73. [44] Madrusov E, Houng A, Klein E, Leonard EF. Membrane-based cell affinity

[45] Malmstadt N, Yager P, Hoffman AS, Stayton PS. A Smart Microfluidic Affinity Chromatography Matrix Composed of Poly(*N*-isopropylacrylamide)-Coated Beads.

[46] Kruger J, Singh K, O'Neill A, Jackson C, Morrison A, O'Brien P. Development of a microfluidic device for fluorescence activated cell sorting. J. Micromech. Microeng.

[47] Dittrich PS, Schwille P. An Integrated Microfluidic System for Reaction, High-Sensitivity Detection, and Sorting of Fluorescent Cells and Particles. Anal. Chem. 2003:

[48] Chung TD, Kim HC. Recent advances in miniaturized microfluidic flow cytometry for

chromatography to retrieve viable cells. Biotechnol Prog 1995: (11) 208-213

[38] Mohamed H, McCurdy LD, Szarowski DH, Duva S, Turner JN, Caggana M.

cancer and its impact on survival. Br Jr Cancer 2003: (88) 574-587.

Treatment 2004: (89) 207-213.

device. PNAS 2009: (106) 10 3970-3975.

[39] IEEE Trans. NanoBio. Sci. 2004: (3) 251-256.

Sorter*. Anal. Chem.* 2002: 74 (11) 2451–2457.

Anal. Chem 2003: 75 (13) 2943-2949.

clinical use. Electrophoresis 2007: (28) 4511–4520.

chip 2005: (5) 30-37.

2002: (12) 486–494.

(75) 5767-5774.

[35] Coventor Inc. Http://coventor.com/ (accessed 29 April 2012).

Biotechnology and Bioengineering 2005; 91(7) 817-826.

[36] ANSYS. Http://ansys.com/ (accessed 29 April 2012). [37] COMSOL. Http://comsol.com/ (accessed 29 April 2012).


Prostate 1994: (25) 249-265.

Biochem. 1998: (70) 172-180.

Biochem, Suppl 1999: (32-33) 183-191.

Immunol Methods 2007: 318 (1–2) 1–5.

128.

microdevices.. Electrophoresis 2000: (*21)* 81-90.

substrates. Anal Chem. 1997: (69) 2626-2630.

Hertfordshire: Research Studies Press LTD: 2000.

transformation. J Cell Biochem. 1992: (49) 357-365.

tumorigenesis. Curr Opin Genet Dev. 2001: (11) 41-47.

[12] Chou C, Austin RH, Bakajin O, Tegenfeldt JO, Castelino JA, Chan SS, Cox EC, Craighead, H, Darnton N, Duke T, Han J, Turner S. Sorting biomolecules with

[13] Lee TMH, Hsing I, Lao AIK, Carles MC. A miniaturized DNA amplifier: its application

[14] Simpson JL, Elias S. Isolating fetal cells from maternal blood: Advances in prenatal

[15] McCormick RM, Nelson RJ, Alonso-Amigo MG, Benvegnu DJ, Hooper HH. Microchannel electrophoretic separations of DNA in injection-molded plastic

[16] Lagally ET, Medintz I, Mathies RA. Single-molecule DNA amplification and analysis in

[17] Koch M, Evans A, Brunnschweiler A. Microfluidic Technology and Applications.

[21] Schaeffer CW, Partin AW, Isaacs WB, Coffey DS, Isaacs JT. Molecular and cellular changes associated with the acquisition of metastatic ability by prostatic cancer cells.

[22] Pienta KJ, Coffey DS. Nuclear-cytoskeletal interactions: evidence for physical connections between the nucleus and cell periphery and their alteration by

[23] Nickerson JA. Nuclear dreams: the malignant alteration of nuclear architecture. J Cell

[24] Pawlak G, Helfman D M. Cytoskeletal changes in cell transformation and

[25] Konety BR, Getzenberg RH. Nuclear structural proteins as biomarkers of cancer. J Cell

[26] Backman V, Wallace MB, Perelman LT, Arendt JT, Gurjar R, Muller MG, Zhang Q, Zonios G, Kline E, McGilligan JA, Shapshay S, Valdez T, Badizadegan K, Crawford JM, Fitzmaurice M, Kabani S, Levin HS, Seiler M, Dasari RR, Itzkan I, Van Dam J, Feld MS,

[27] Zola H, Swart B, Banham A, Barry S, Beare A, Bensussan A, Boumsell L, D Buckley C, Bühring HJ, Clark G, Engel P, Fox D, Jin BQ, Macardle PJ, Malavasi F, Mason D, Stockinger H, Yang X. CD molecules 2006--human cell differentiation molecules. J

[28] Went P, Lugli A., Meier S , Bundi M, Mirlacher M, Sauter G, Dirnhofer S. Frequent EpCam Protein Expression in Human Carcinomas. Human Pathology 2004: (35) 122-

[29] Gastl G, Spizzo G, Obrist P, Dunser M, Mikuz G. Ep-CAM overexpression in breast

cancer as a predictor of survival. The Lancet 2000: (356) 1981-1082.

McGillican T. Detection of preinvasive cancer cells. Nature 2000: (406) 35-36.

[18] Saliterman SS. BioMEMS and Medical Microdevices. Bellingham: SPIE Press; 2006. [19] Bunn HF, Aster JC. Pathophysiology of Blood Disorders. Boston: McGrawy Hill: 2010.

[20] Stuart M, Nagel R. Sickle-cell disease. The Lancet 2004: 364 (9442) 1343-1360

in traditional chineses medicine. Anal Chem, 2000: (72) 4242-4247.

an integrated microfluidic device. Anal Chem. 2001: (73) 565-570.

diagnosis through molecular technology. JAMA 1993: (270) 2357-2361.

	- [49] Miyake R, Ohki H, Yamazaki I, Takagi T. Investigation of sheath flow chambers for flow cytometers (micromachined flow chamber with low pressure loss). JSME Int. J. B 1997: 40) 106–13.

Use of Microfluidic Technology for Cell Separation 225

[68] Chen YC, Lou X, Ingram P, Yoon E. The 15th International Conference on Miniaturized Systems for Chemistry and Life Sciences: conference proceedings, October 2-6, 2011,

[69] Englert DL, Manson MD, Jayaraman A. Flow-Based Microfluidic Device for Quantifying Bacterial Chemotaxis in Stable, Competing Gradients. Applied and

[70] Walker GM, Sai J, Richmond A, Stremler M, Chung CY, Wikswo JP. Effects of flow and diffusion on chemotaxis studies in a microfabricated gradient generator. Lab on a Chip

[71] MacDonald MP, Spalding GC, Dholakia K. Microfluidic sorting in an optical lattice.

[72] Kovac JR, Voldman J. Intuitive, Image-Based Cell Sorting Using Optofluidic Cell

[73] Shirasaki Y, Tanaka J, Makazu H, Tashiro K, Shoji S, Tsukita S, Funatsu T. On-Chip Cell Sorting System Using Laser-Induced Heating of a Thermoreversible Gelatin Polymer to

[74] Lin CC, Chen A, Kin CH. Microfluidic cells counter/sorter utilizing multiple particle tracing technique and optically switching approach. Biomed Microdevices 2008: (10) 55-

[75] Hwang H, Park JK. Optoelectrofluidic platforms for chemistry and biology. Lab on a

[76] Wang MM, Tu E, Raymond DE, Yang MJ, Zhang H, Hagen N, Dees B, Mercer EM, Forster AH, Kariv I, Marchand PJ, Butler W. Nature Biotechnology 2004: (23) 83-87. [77] Pommer MS, Zhang Y, Keerthi N, Chen D, Thomson JA, Meinhard CD, Soh, HT. Dielectrophoretic separation of platelets from diluted whole blood in microfluidic

[78] Hu X, Bessette PH, Qian J, Meinhard CD, Daugherty PS, Soh HT. Marker-specific sorting of rare cells using dielectrophoresis. PNAS 2005: (102) 44 15757-15761. [79] Vahey MD, Voldman J. An Equilibrium Method for Continuous-Flow Cell Sorting

[80] Lapizco-Encinas BH, Simmons BA, Cummings EB, Fintschenko Y. Dielectrophoretic Concentration and Separation of Live and Dead Bacteria in an Array of Insulators. Anal

[81] Morijiri T, Sunahiro S, Senaha M, Yamada M, Seki M. Sedimentation pinched-flow fractionation for size- and density-based particle sorting in microchannels. Microfluid

[82] Wilding P, Kricka LJ, Cheng J, Hvichia G, Shoffner MA, Fortina P. Integrated cell isolation and polymerase chain reaction analysis using silicon microfilter chambers.

[83] Yuen PK, Kricka LJ, Fortina P, Panaro PJ, Sakazume T, Wilding P. Microchip module for blood sample preparation and nucleic acid amplification reactions. Genome Res

Seattle, Washington.

2005: (5) 611-618.

63.

Nature 2003: (426) 421-424.

Chip 2011: (11) 33-47.

Chem 2004: (76) 1571-1579.

Nanofluid 2011: (11) 105-110.

Anal Biochem 1998: (257) 95-200.

2001: (11) 405-412.

Enviromental Microbiology 2009: (75) 4557–4564

Sorting. Anal. Chem. 2007: (79) 9321-9330.

Control Flow. Anal Chem 2006: (78) 695-701.

channels. Electrophoresis 2008: (29) 1213-1218.

Using Dielectrophoresis. Anal Chem 2008: (80) 3135-3143.


[68] Chen YC, Lou X, Ingram P, Yoon E. The 15th International Conference on Miniaturized Systems for Chemistry and Life Sciences: conference proceedings, October 2-6, 2011, Seattle, Washington.

224 Blood Cell – An Overview of Studies in Hematology

Biotechnol. Prog. 2002: (18) 1439-1442.

Chem 2008: (392) 1317–1324.

San Diego, California, USA.

Biomed Microdevices 2006: (8) 299–308.

microspheres. Microfluid Nanofluid 2009: (6) 647–660.

magnetic arrays. PNAS 2010: (107) 33 14525-14529.

105 (47) 18165–18170.

196.

1997: 40) 106–13.

(81) 5188–5196

[49] Miyake R, Ohki H, Yamazaki I, Takagi T. Investigation of sheath flow chambers for flow cytometers (micromachined flow chamber with low pressure loss). JSME Int. J. B

[50] Huh D, Gu W, Kamotani Y, Grotberg JB, Takayama S. Microfluidics for flow cytometric

[51] Altendorf E, Zebert D, Holl M, Yager P. Differential blood cell counts obtained using a

[52] Fu AY, Spence C, Scherer A, Arnold FH, Quake SR. A microfabricated fluorescence-

[53] Oakey J, Allely J, Marr DWM. Laminar-Flow-Based Separations at the Microscale.

[54] Johansson L, Nikolajeff F, Johansson S, Thorslund S. On-Chip Fluorescence-Activated Cell Sorting by an Integrated Miniaturized Ultrasonic Transducer. Anal. Chem. 2009:

[55] Chun H, Chung TD, Kim HC, Cytometry and Velocimetry on a Microfluidic Chip

[57] Han KH, Frazier B. Paramagnetic capture mode magnetophoretic microseparator for

[58] Qu BY,Wu ZY, Fang F, Bai ZM, Yang DZ, Xu SK. A glass microfluidic chip for continuous blood cell sorting by a magnetic gradient without labeling. Anal Bioanal

[59] Adams JD, Kimb U, Sohb HT. Multitarget magnetic activated cell sorter. PNAS 2008:

[60] Xia N, Hunt TP, Mayers BT, Alsberg E*,* Whitesides *GM,* Westervelt RM, Ingber DE. Combined microfluidic-micromagnetic separation of living cells in continuous flow.

[61] Modak N, Datta A, Ganguly R. Cell separation in a microfluidic channel using magnetic

[62] Lee H, Purdon AM, Westervelta RM. Manipulation of biological cells using a

[63] Salibaa AE, Saiasa L, Psycharia E, Minca N, Simonb D, Bidardc FC, Mathiotd C, Piergac JY, Fraisierf V, Salamerof J, Saadag V, Faraceg F, Vielhg P, Malaquina L, Jean-Louis Viovya JL. Microfluidic sorting and multimodal typing of cancer cells in self-assembled

[64] Kose AR, Koser H. Ferrofluid mediated nanocytometry. Lab on a Chip 2012: (12) 190-

[67] Agrawal N, Toner M, Irimia D. Twelfth International Conference on Miniaturized Systems for Chemistry and Life Sciences: conference proceedings, October 12-16, 2008,

[65] Li, D Electrokinetics in microfluidics, 1st edition, vol. 2, Elsevier, Amsterdam (2004). [66] Al-Abboodi A, Tjeung R, Doran P, Yeo L, Friend J, Chan P. Microfluidic chip containing

porous gradient for chemotaxis study. Proc. of SPIE 2011: (8204) H1-H6

microelectromagnet matrix. Applied Physics Letters 2004: (85) 1063-1065.

analysis of cells and particles. Physiol. Meas. 2005: (26) R73–R98.

activated cell sorter. Nature Biotechnology 1999: (17) 1109-1111

microchannel based flow cytometer. Transducers 1997: (97) 531–534.

Using Polyelectrolytic Salt Bridges. Anal. Chem. 2005: (77) 2490-2495. [56] Pamme N. Magnetism and microfluidics. Lab on a chip 2006: (6) 24-38.

high efficiency blood cell separations. Lab on a Chip 2006: (6) 265-273

	- [84] Panaro NJ, Lou XJ, Fortina P, Kicka LJ, Wilding P. Micropillar array chip for integrated white blood cell isolation and PCR. Biomed Eng 2005: (21) 157-162.

**Section 3** 

**Applications in Haematology** 


**Applications in Haematology** 

226 Blood Cell – An Overview of Studies in Hematology

Conference, vol1, pp. 1-4, 2005.

May 20-24, 2007, Santa Clara, California.

14784.

237.

899.

[84] Panaro NJ, Lou XJ, Fortina P, Kicka LJ, Wilding P. Micropillar array chip for integrated

[85] Cabodi M, ChenYF, Turner SWP, Craighead HG, Austin RH. Continuous separation of biomolecules by the laterally asymmetric diffusion array with out-of-plane sample

[86] Huang LR. Cox EC, Austin RH, Sturm JC. Continuous Particle Separation Through

[87] Davis JA, Inglis DW, Morton KJ, Lawrence DA, Huang LR, Chou SY, Sturm JC, Austin RH. Deterministic hydrodynamics: Taking blood apart. PNAS 2006: 103 (40) 14779–

[88] Mohamed H, McCurdy LD, Szarowski DH, Duva S, Turner JN, Caggana M. Development of a Rare-Cell Fractionation Device: Application for Cancer Detection.

[89] H. Mohamed, M. Murray, J. N. Turner, and M. Caggana, "Circulating tumor cells: capture with a micromachined device," Proceedings of the 2005 NSTI Bio Nano

[90] Mohamed H, Murray M, Turner JN, Caggana M. Isolation of tumor cells using size and

[91] H. Mohamed, J. N. Turner, and M. Caggana. Bio Nano Conference: conference proceedings, May 7-11, 2006, Hynes Convention Center, Boston Massachusetts. [92] Mohamed H, Turner JN, Caggana M. Bio Nano Conference: conference proceedings,

[93] Murthy SK, Sethu P,Vunjak-Novakovic G, Toner M, Radisic M. Size-based microfluidic enrichment of neonatal rat cardiac cell populations. Biomed Microdevices 2006: (8) 231–

[94] Huang R, Barber TA, Schmidt MA, Tompkins RG, Toner M, Bianchi DW, Kapur R, Flejter WL. A microfluidics approach for the isolation of nucleated red blood cells (fNRBCs) from the peripheral blood of pregnant women. Prenat Diagn. 2008: (28) 892–

deformation. Journal of Chromatography A 2009: (1216) 8289–8295.

white blood cell isolation and PCR. Biomed Eng 2005: (21) 157-162.

Deterministic Lateral Displacement. Science 2004: (304) 987-990.

IEEE Transactions on NanoBioscience 2004: (3) 251-256.

injection. Electrophoresis 2002: (23) 3496–3503.

**Chapter 12** 

© 2012 Shrivastav and Singh, licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2012 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution,

© 2012 Shrivastav and Singh, licensee InTech. This is a paper distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use,

distribution, and reproduction in any medium, provided the original work is properly cited.

and reproduction in any medium, provided the original work is properly cited.

**Tigers Blood:** 

A.B. Shrivastav and K.P. Singh

http://dx.doi.org/10.5772/50360

**1. Introduction** 

Additional information is available at the end of the chapter

member of forest ecology (Shrivastav, 2001).

may reflect the physiological disorders (Harvey, 1997).

**Haematological and Biochemical Studies** 

Tiger (*Panthera tigris tigris*) population in their historic ranges is critically endangered owing to habitat destructions, ruthless poaching and retaliatory killing. The tiger population now remains in few thousands located in about 150 fragments in 13 countries (Karanth and Gopal, 2005). However, declination is also associated with health related problems such as nutritional deficiencies and infectious diseases (Prater, 2005). Therefore, health monitoring and scientific health management, disease diagnosis and treatment should be made mandatory for conservation of wildlife as the tiger is a key stone species and important

Haematological and biochemical studies are important tool for health evaluation and their interpretations to know the status of physiological functions of various organs. The concentration of biochemical constituents in tissues as well as in body fluid is fixed and during adverse conditions, it may be elevated or decreased (Douglas and Nelson, 1991). However, qualitative and quantitative analysis of corpuscles and chemical constituents of plasma or serum are closely linked with functional unit of the cell and their assessments

Nevertheless, several factors involved to transmit infectious diseases either mechanically or biologically through contaminated water, food or vectors (Lice, Flea, Ticks and Mites) and the pathogens may alter the normal physiology (Shah, 1983). Viral, bacterial and parasitic diseases are very common in tigers which can affect the haematological and biochemical normal values (Rao and Acharyjo, 2002). Types of anaemia and significant blood loss may be estimated through complete blood count (CBC) and physiological function of different organs by biochemical parameters (Jain, 1986). Qualitative and quantitative reduction in the blood commonly observed in captive felid particularly in cubs those maintaining on milk alone. The values of liver function test, elevated on repeated immobilization by sedative
