Skip to main content
Log in

Cellular imaging using phase holographic microscopy: for the study of pathophysiology of red blood cells and human umbilical vein endothelial cells

  • Regular Paper
  • Published:
Journal of Visualization Aims and scope Submit manuscript

Abstract

Digital holographic microscopy (DHM) for phase-contrast imaging is employed for live cell imaging to study pathophysiology of red blood cells (RBCs) and human umbilical vein endothelial cells (HUVECs). The morphology and membrane fluctuation of healthy RBCs are investigated by adapting post-image processing which automatically quantify the morphological properties of RBCs, such as perimeter, mean diameter, cell volume, cell thickness and membrane fluctuation. Moreover, morphological variation of malaria parasite-infected RBCs and hypertonic-shocked RBCs was observed. As another biological application, HUVECs cultured on collagen films and their abnormal deformations caused by collagenase are observed using DHM to understand the role of extracellular matrix (ECM) structure on atheroprotective effect and endothelial dysfunction.

Graphical Abstract

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  • Byun H, Hillman TR, Higgins JM, Diez-Silva M, Peng ZL, Dao M et al (2012) Optical measurement of biomechanical properties of individual erythrocytes from a sickle cell patient. Acta Biomater 8:8–4130

    Article  Google Scholar 

  • Cuche E, Marquet P, Depeursinge C (1999) Simultaneous amplitude-contrast and quantitative phase-contrast microscopy by numerical reconstruction of Fresnel off-axis holograms. Appl Opt 38:6994–7001

    Article  Google Scholar 

  • Herraez MA, Burton DR, Lalor MJ, Gdeisat MA (2002) Fast two-dimensional phase-unwrapping algorithm based on sorting by reliability following a noncontinuous path. Appl Opt 41:7437–7444

    Article  Google Scholar 

  • Kemper B, Bauwens A, Vollmer A, Ketelhut S, Langehanenberg P, Muthing J et al (2010) Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy. J Biomed Opt 15:036009

    Article  Google Scholar 

  • Mann CJ, Yu LF, Lo CM, Kim MK (2005) High-resolution quantitative phase-contrast microscopy by digital holography. Opt Express 13:8693–8698

    Article  Google Scholar 

  • Marquet P, Rappaz B, Magistretti PJ, Cuche E, Emery Y, Colomb T et al (2005) Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy. Opt Lett 30:468–470

    Article  Google Scholar 

  • McLaren CE, Brittenham GM, Hasselblad V (1987) Statistical and graphical evaluation of erythrocyte volume distributions. Am J Physiol 252:H857–H866

    Google Scholar 

  • Nomarski G (1955) Differential microinterferometer with polarized waves. J Phys Radium 16:6–13

    Google Scholar 

  • Park YK, Diez-Silva M, Popescu G, Lykotrafitis G, Choi WS, Feld MS et al (2008) Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum. Proc Natl Acad Sci USA 105:13730–13735

    Article  Google Scholar 

  • Park Y, Best CA, Badizadegan K, Dasari RR, Feld MS, Kuriabova T et al (2010) Measurement of red blood cell mechanics during morphological changes. Proc Natl Acad Sci USA 107:6731–6736

    Article  Google Scholar 

  • Park Y, Best CA, Kuriabova T, Henle ML, Feld MS, Levine AJ et al (2011) Measurement of the nonlinear elasticity of red blood cell membranes. Phys Review E 83:051925

    Article  Google Scholar 

  • Popescu G, Ikeda T, Best CA, Badizadegan K, Dasari RR, and Feld MS (2005) Erythrocyte structure and dynamics quantified by Hilbert phase microscopy. J Biomed Opt 10

  • Popescu G, Ikeda T, Dasari RR, Feld MS (2006) Diffraction phase microscopy for quantifying cell structure and dynamics. Opt Lett 31:7–775

    Article  Google Scholar 

  • Popescu G, Park Y, Choi W, Dasari RR, Feld MS, Badizadegan K (2008) Imaging red blood cell dynamics by quantitative phase microscopy. Blood Cells Mol Dis 41:10–16

    Article  Google Scholar 

  • Rappaz B, Marquet P, Cuche E, Emery Y, Depeursinge C, Magistretti PJ (2005) Measurement of the integral refractive index and dynamic cell morphometry of living cells with digital holographic microscopy. Opt Express 13:9361–9373

    Article  Google Scholar 

  • Seo KW, Choi YS, Seo ES, Lee SJ (2012) Aberration compensation for objective phase curvature in phase holographic microscopy. Opt Lett 37:8–4976

    Article  Google Scholar 

  • Seo ES, Seo KW, Park J, Lee TG, and Lee SJ (2014) Study on the deformation of endothelial cells using a bio-inspired in vitro disease model. Microvasc Res, http://dx.doi.org/10.1016/j.mvr.2014.02.003

  • Yamaguchi I, Zhang T (1997) Phase-shifting digital holography. Opt Lett 22:1268–1270

    Article  Google Scholar 

  • Yu LF, Kim MK (2005) Wavelength-scanning digital interference holography for tomographic three-dimensional imaging by use of the angular spectrum method. Opt Lett 30:2092–2094

    Article  Google Scholar 

  • Zernike F (1942a) Phase contrast, a new method for the microscopic observation of transparent objects. Physica 9:686–698

    Article  Google Scholar 

  • Zernike F (1942b) Phase contrast, a new method for the microscopic observation of transparent objects part II. Physica 9:974–980

    Article  Google Scholar 

  • Zhang T, Yamaguchi I (1998) Three-dimensional microscopy with phase-shifting digital holography. Opt Lett 23:3–1221

    Google Scholar 

Download references

Acknowledgments

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (No. 2008-0061991).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Sang Joon Lee.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Seo, K.W., Seo, E. & Lee, S.J. Cellular imaging using phase holographic microscopy: for the study of pathophysiology of red blood cells and human umbilical vein endothelial cells. J Vis 17, 235–244 (2014). https://doi.org/10.1007/s12650-014-0200-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12650-014-0200-y

Keywords

Navigation