Abstract

We have proposed and demonstrated a very sensitive volume measurement scheme for a live cell with a quantitative phase microscopy (QPM) utilizing auto-focusing and numerical edge detection schemes. An auto-focusing technique with two different focus measures is applied to find the focus dependent errors in our live cell volume measurement system. The volume of a polystyrene bead sample with 3 μm diameter has been measured for the validity test of our proposed method. We have shown that a small displacement of an object from its focusing position can cause a large volume error. A numerical edge detection technique is also used to accurately resolve the boundary between a cell and its suspension medium. We have applied this method to effectively suppress errors by the surrounding medium of a single red blood cell (RBC).

© 2009 Optical Society of America

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    [CrossRef]
  2. C. L. Curl, C. J. Bellair, P. J. Harris, B. E. Allman, A. Roberts, K. A. Nugent, and L. M. D. Delbridge, "Single Cell Volume Measurement by Quantitative Phase Microscopy (QPM): A Case Study of Erythrocyte Morphology," Cell Physiol Biochem 17, 193-200 (2006)
    [CrossRef] [PubMed]
  3. H. Satoh, L. M. D. Delbridge, L. A. Blatter, and D. M. Bers, "Volume Relationship in Cardiac Myocytes Studied with Confocal Microscopy and Membrane Capacitance Measurements: Species-Dependence and Developmental Effects," Biophys. J. 70, 1494-1504 (1996)
    [CrossRef] [PubMed]
  4. Y. E. Korchev, J. Gorelik, M. J. Lab, E. V. Sviderskaya, C. L. Johnston, C. R. Coombes, I. Vodyanoy, and C. R. W. Edwards, "Cell volume Measurement using Scanning Ion Conductance Microscopy," Biophys. J. 78, 451-457 (2000)
    [CrossRef] [PubMed]
  5. G. S and F. W, "Amiloride-sensitive Na+/H+ exchange in human neutrophils: mechanism of activation by chemotactic factors," Biochem. Biophys. Res. Comm. 122, 755-762 (1984)
    [CrossRef]
  6. Y. R. Kim and L. Ornstein, "Isovolumetric Sphering of Erythrocytes for more accurate and precise cell volume measurement by flow Cytometry," Cytometry 3, 419-427 (1983)
    [CrossRef] [PubMed]
  7. P. J. Harris, J. Y. Chatton, P. H. Tran, P. M. Bungay, and K. R. Spring, "pH, morphology, and diffusion in lateral intercellular spaces of epithelial cell monolayers," Am. J. Physiol. 266, C73-C80 (1994)
    [PubMed]
  8. C. M. Lee, "Measurement of volume injected into individual cells by quantitative fluorescence microscopy," J. Cell Sci 94, 443-447 (1989)
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    [CrossRef]
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    [PubMed]
  11. J. Farinas, M. Kneen, M. Moore, and Verkman, "Plasma membrane water permeability of cultured cells and epithelia measured by light microscopy with spatial filtering," J. Gen. Physiol. 110, 283-296 (1997)
    [CrossRef] [PubMed]
  12. G.  Popescu, L. P.  Deflores, J. C.  Vaughan, K.  Badizadegan, H.  Iwai, R. R.  Dasari, and M. S.  Feld, "Fourier phase microscopy for investigation of biological structures and dynamics," Opt. Lett.  29, 2503-2505 (2004)
    [CrossRef] [PubMed]
  13. N. Lue, W. Choi, and G. Popescu, T. Ikeda, R. R. Dasari, K. Badizadegan, and M. S. Feld, "Quantitative phase imaging of live cells using fast Fourier phase microscopy," Appl. Opt. 46, 1836-1842 (2007)
    [CrossRef] [PubMed]
  14. T.  Ikeda, G.  Popescu, R. R.  Dasari, and M. S.  Feld, "Hilbert phase microscopy for investigating fast dynamics in transparent systems," Opt. Lett.  30, 1165-1168 (2005)
    [CrossRef] [PubMed]
  15. G.  Popescu, T.  Ikeda, R. R.  Dasari, and M. S.  Feld, "Diffraction phase microscopy for quantifying cell structure and dynamics," Opt. Lett.  31, 775-777 (2006)
    [CrossRef] [PubMed]
  16. Y. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, "Diffraction phase and fluorescence microscopy," Opt. Express 14, 8263-8268 (2006)
    [CrossRef] [PubMed]
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  19. C. Fang-Yen, S. Oh, Y. Park, W. Choi, S. Song, H. S. Seung, R. R. Dasari, and M. S. Feld, "Imaging voltage-dependent cell motions with heterodyne Mach-Zehnder phase microscopy," Opt. Lett. 32, 1572-1574 (2007)
    [CrossRef] [PubMed]
  20. F. C. A. Groen, I. T. Young, and G. Ligthart, "A Comparison of Different Focus Functions for Use in Autofocus Algorithms," Cytometry 6, 81-91 (1985)
    [CrossRef] [PubMed]
  21. P. Langehanenberg, B. Kemper, D. Dirksen, and G. V. Bally, "Autofocusing in digital holographic phase contrast microscopy on pure phase objects for live cell imaging," Appl. Opt. 47, 176-181 (2008)
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  23. Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, "Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum," PNAS.  105, 13730-13735 (2008)
    [CrossRef] [PubMed]
  24. J. Canny, "A Computational Approach To Edge Detection," IEEE Trans. Pattern Analysis and Machine Intelligence,  8:679-714 (1986).
    [CrossRef]
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    [CrossRef]

2008 (2)

P. Langehanenberg, B. Kemper, D. Dirksen, and G. V. Bally, "Autofocusing in digital holographic phase contrast microscopy on pure phase objects for live cell imaging," Appl. Opt. 47, 176-181 (2008)
[CrossRef]

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, "Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum," PNAS.  105, 13730-13735 (2008)
[CrossRef] [PubMed]

2007 (2)

2006 (3)

G.  Popescu, T.  Ikeda, R. R.  Dasari, and M. S.  Feld, "Diffraction phase microscopy for quantifying cell structure and dynamics," Opt. Lett.  31, 775-777 (2006)
[CrossRef] [PubMed]

Y. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, "Diffraction phase and fluorescence microscopy," Opt. Express 14, 8263-8268 (2006)
[CrossRef] [PubMed]

C. L. Curl, C. J. Bellair, P. J. Harris, B. E. Allman, A. Roberts, K. A. Nugent, and L. M. D. Delbridge, "Single Cell Volume Measurement by Quantitative Phase Microscopy (QPM): A Case Study of Erythrocyte Morphology," Cell Physiol Biochem 17, 193-200 (2006)
[CrossRef] [PubMed]

2005 (3)

2004 (1)

2000 (1)

Y. E. Korchev, J. Gorelik, M. J. Lab, E. V. Sviderskaya, C. L. Johnston, C. R. Coombes, I. Vodyanoy, and C. R. W. Edwards, "Cell volume Measurement using Scanning Ion Conductance Microscopy," Biophys. J. 78, 451-457 (2000)
[CrossRef] [PubMed]

1999 (1)

L. Allansson, S. Khatibi, T. Gustavsson, F. Blomstrand, T. Olsson, and E. Hansson, "Single cell volume estimation by three dimensional wide field microscopy applied to astroglial primary cultures," J. Neurosci. Meth. 93,1-11 (1999)
[CrossRef]

1998 (1)

1997 (1)

J. Farinas, M. Kneen, M. Moore, and Verkman, "Plasma membrane water permeability of cultured cells and epithelia measured by light microscopy with spatial filtering," J. Gen. Physiol. 110, 283-296 (1997)
[CrossRef] [PubMed]

1996 (1)

H. Satoh, L. M. D. Delbridge, L. A. Blatter, and D. M. Bers, "Volume Relationship in Cardiac Myocytes Studied with Confocal Microscopy and Membrane Capacitance Measurements: Species-Dependence and Developmental Effects," Biophys. J. 70, 1494-1504 (1996)
[CrossRef] [PubMed]

1994 (1)

P. J. Harris, J. Y. Chatton, P. H. Tran, P. M. Bungay, and K. R. Spring, "pH, morphology, and diffusion in lateral intercellular spaces of epithelial cell monolayers," Am. J. Physiol. 266, C73-C80 (1994)
[PubMed]

1992 (1)

K. HK, O. ER, S. P, and K. C, "Methods for determination of cell volume in tissue culture," Can J. Physiol Pharmacol 70, S323-S333 (1992)
[CrossRef]

1990 (1)

T. Nakahari, M. Murakami, H. Yoshida, M. Miyamoto, Y. Sohma, and Y. Imai, "Decrease in rat submandibular acinar cell volume during Ach stimulation," Am. J. Physiol. 258, G878-G886 (1990)
[PubMed]

1989 (1)

C. M. Lee, "Measurement of volume injected into individual cells by quantitative fluorescence microscopy," J. Cell Sci 94, 443-447 (1989)
[PubMed]

1986 (1)

J. Canny, "A Computational Approach To Edge Detection," IEEE Trans. Pattern Analysis and Machine Intelligence,  8:679-714 (1986).
[CrossRef]

1985 (1)

F. C. A. Groen, I. T. Young, and G. Ligthart, "A Comparison of Different Focus Functions for Use in Autofocus Algorithms," Cytometry 6, 81-91 (1985)
[CrossRef] [PubMed]

1984 (1)

G. S and F. W, "Amiloride-sensitive Na+/H+ exchange in human neutrophils: mechanism of activation by chemotactic factors," Biochem. Biophys. Res. Comm. 122, 755-762 (1984)
[CrossRef]

1983 (1)

Y. R. Kim and L. Ornstein, "Isovolumetric Sphering of Erythrocytes for more accurate and precise cell volume measurement by flow Cytometry," Cytometry 3, 419-427 (1983)
[CrossRef] [PubMed]

Allansson, L.

L. Allansson, S. Khatibi, T. Gustavsson, F. Blomstrand, T. Olsson, and E. Hansson, "Single cell volume estimation by three dimensional wide field microscopy applied to astroglial primary cultures," J. Neurosci. Meth. 93,1-11 (1999)
[CrossRef]

Allman, B. E.

C. L. Curl, C. J. Bellair, P. J. Harris, B. E. Allman, A. Roberts, K. A. Nugent, and L. M. D. Delbridge, "Single Cell Volume Measurement by Quantitative Phase Microscopy (QPM): A Case Study of Erythrocyte Morphology," Cell Physiol Biochem 17, 193-200 (2006)
[CrossRef] [PubMed]

Badizadegan, K.

Bally, G. V.

P. Langehanenberg, B. Kemper, D. Dirksen, and G. V. Bally, "Autofocusing in digital holographic phase contrast microscopy on pure phase objects for live cell imaging," Appl. Opt. 47, 176-181 (2008)
[CrossRef]

Bellair, C. J.

C. L. Curl, C. J. Bellair, P. J. Harris, B. E. Allman, A. Roberts, K. A. Nugent, and L. M. D. Delbridge, "Single Cell Volume Measurement by Quantitative Phase Microscopy (QPM): A Case Study of Erythrocyte Morphology," Cell Physiol Biochem 17, 193-200 (2006)
[CrossRef] [PubMed]

Bers, D. M.

H. Satoh, L. M. D. Delbridge, L. A. Blatter, and D. M. Bers, "Volume Relationship in Cardiac Myocytes Studied with Confocal Microscopy and Membrane Capacitance Measurements: Species-Dependence and Developmental Effects," Biophys. J. 70, 1494-1504 (1996)
[CrossRef] [PubMed]

Blatter, L. A.

H. Satoh, L. M. D. Delbridge, L. A. Blatter, and D. M. Bers, "Volume Relationship in Cardiac Myocytes Studied with Confocal Microscopy and Membrane Capacitance Measurements: Species-Dependence and Developmental Effects," Biophys. J. 70, 1494-1504 (1996)
[CrossRef] [PubMed]

Blomstrand, F.

L. Allansson, S. Khatibi, T. Gustavsson, F. Blomstrand, T. Olsson, and E. Hansson, "Single cell volume estimation by three dimensional wide field microscopy applied to astroglial primary cultures," J. Neurosci. Meth. 93,1-11 (1999)
[CrossRef]

Bungay, P. M.

P. J. Harris, J. Y. Chatton, P. H. Tran, P. M. Bungay, and K. R. Spring, "pH, morphology, and diffusion in lateral intercellular spaces of epithelial cell monolayers," Am. J. Physiol. 266, C73-C80 (1994)
[PubMed]

Canny, J.

J. Canny, "A Computational Approach To Edge Detection," IEEE Trans. Pattern Analysis and Machine Intelligence,  8:679-714 (1986).
[CrossRef]

Chatton, J. Y.

P. J. Harris, J. Y. Chatton, P. H. Tran, P. M. Bungay, and K. R. Spring, "pH, morphology, and diffusion in lateral intercellular spaces of epithelial cell monolayers," Am. J. Physiol. 266, C73-C80 (1994)
[PubMed]

Choi, W.

Colomb, T.

Coombes, C. R.

Y. E. Korchev, J. Gorelik, M. J. Lab, E. V. Sviderskaya, C. L. Johnston, C. R. Coombes, I. Vodyanoy, and C. R. W. Edwards, "Cell volume Measurement using Scanning Ion Conductance Microscopy," Biophys. J. 78, 451-457 (2000)
[CrossRef] [PubMed]

Cuche, E.

Curl, C. L.

C. L. Curl, C. J. Bellair, P. J. Harris, B. E. Allman, A. Roberts, K. A. Nugent, and L. M. D. Delbridge, "Single Cell Volume Measurement by Quantitative Phase Microscopy (QPM): A Case Study of Erythrocyte Morphology," Cell Physiol Biochem 17, 193-200 (2006)
[CrossRef] [PubMed]

Dasari, R. R.

Deflores, L. P.

Delbridge, L. M. D.

C. L. Curl, C. J. Bellair, P. J. Harris, B. E. Allman, A. Roberts, K. A. Nugent, and L. M. D. Delbridge, "Single Cell Volume Measurement by Quantitative Phase Microscopy (QPM): A Case Study of Erythrocyte Morphology," Cell Physiol Biochem 17, 193-200 (2006)
[CrossRef] [PubMed]

H. Satoh, L. M. D. Delbridge, L. A. Blatter, and D. M. Bers, "Volume Relationship in Cardiac Myocytes Studied with Confocal Microscopy and Membrane Capacitance Measurements: Species-Dependence and Developmental Effects," Biophys. J. 70, 1494-1504 (1996)
[CrossRef] [PubMed]

Depeursinge, C.

Diez-Silva, M.

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, "Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum," PNAS.  105, 13730-13735 (2008)
[CrossRef] [PubMed]

Dirksen, D.

P. Langehanenberg, B. Kemper, D. Dirksen, and G. V. Bally, "Autofocusing in digital holographic phase contrast microscopy on pure phase objects for live cell imaging," Appl. Opt. 47, 176-181 (2008)
[CrossRef]

Edwards, C. R. W.

Y. E. Korchev, J. Gorelik, M. J. Lab, E. V. Sviderskaya, C. L. Johnston, C. R. Coombes, I. Vodyanoy, and C. R. W. Edwards, "Cell volume Measurement using Scanning Ion Conductance Microscopy," Biophys. J. 78, 451-457 (2000)
[CrossRef] [PubMed]

Emery, Y.

Fang-Yen, C.

Farinas, J.

J. Farinas, M. Kneen, M. Moore, and Verkman, "Plasma membrane water permeability of cultured cells and epithelia measured by light microscopy with spatial filtering," J. Gen. Physiol. 110, 283-296 (1997)
[CrossRef] [PubMed]

Feld, M. S.

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, "Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum," PNAS.  105, 13730-13735 (2008)
[CrossRef] [PubMed]

C. Fang-Yen, S. Oh, Y. Park, W. Choi, S. Song, H. S. Seung, R. R. Dasari, and M. S. Feld, "Imaging voltage-dependent cell motions with heterodyne Mach-Zehnder phase microscopy," Opt. Lett. 32, 1572-1574 (2007)
[CrossRef] [PubMed]

N. Lue, W. Choi, and G. Popescu, T. Ikeda, R. R. Dasari, K. Badizadegan, and M. S. Feld, "Quantitative phase imaging of live cells using fast Fourier phase microscopy," Appl. Opt. 46, 1836-1842 (2007)
[CrossRef] [PubMed]

Y. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, "Diffraction phase and fluorescence microscopy," Opt. Express 14, 8263-8268 (2006)
[CrossRef] [PubMed]

G.  Popescu, T.  Ikeda, R. R.  Dasari, and M. S.  Feld, "Diffraction phase microscopy for quantifying cell structure and dynamics," Opt. Lett.  31, 775-777 (2006)
[CrossRef] [PubMed]

T.  Ikeda, G.  Popescu, R. R.  Dasari, and M. S.  Feld, "Hilbert phase microscopy for investigating fast dynamics in transparent systems," Opt. Lett.  30, 1165-1168 (2005)
[CrossRef] [PubMed]

G.  Popescu, L. P.  Deflores, J. C.  Vaughan, K.  Badizadegan, H.  Iwai, R. R.  Dasari, and M. S.  Feld, "Fourier phase microscopy for investigation of biological structures and dynamics," Opt. Lett.  29, 2503-2505 (2004)
[CrossRef] [PubMed]

Gorelik, J.

Y. E. Korchev, J. Gorelik, M. J. Lab, E. V. Sviderskaya, C. L. Johnston, C. R. Coombes, I. Vodyanoy, and C. R. W. Edwards, "Cell volume Measurement using Scanning Ion Conductance Microscopy," Biophys. J. 78, 451-457 (2000)
[CrossRef] [PubMed]

Groen, F. C. A.

F. C. A. Groen, I. T. Young, and G. Ligthart, "A Comparison of Different Focus Functions for Use in Autofocus Algorithms," Cytometry 6, 81-91 (1985)
[CrossRef] [PubMed]

Gustavsson, T.

L. Allansson, S. Khatibi, T. Gustavsson, F. Blomstrand, T. Olsson, and E. Hansson, "Single cell volume estimation by three dimensional wide field microscopy applied to astroglial primary cultures," J. Neurosci. Meth. 93,1-11 (1999)
[CrossRef]

Hammer, M.

Hansson, E.

L. Allansson, S. Khatibi, T. Gustavsson, F. Blomstrand, T. Olsson, and E. Hansson, "Single cell volume estimation by three dimensional wide field microscopy applied to astroglial primary cultures," J. Neurosci. Meth. 93,1-11 (1999)
[CrossRef]

Harris, P. J.

C. L. Curl, C. J. Bellair, P. J. Harris, B. E. Allman, A. Roberts, K. A. Nugent, and L. M. D. Delbridge, "Single Cell Volume Measurement by Quantitative Phase Microscopy (QPM): A Case Study of Erythrocyte Morphology," Cell Physiol Biochem 17, 193-200 (2006)
[CrossRef] [PubMed]

P. J. Harris, J. Y. Chatton, P. H. Tran, P. M. Bungay, and K. R. Spring, "pH, morphology, and diffusion in lateral intercellular spaces of epithelial cell monolayers," Am. J. Physiol. 266, C73-C80 (1994)
[PubMed]

Ikeda, T.

Imai, Y.

T. Nakahari, M. Murakami, H. Yoshida, M. Miyamoto, Y. Sohma, and Y. Imai, "Decrease in rat submandibular acinar cell volume during Ach stimulation," Am. J. Physiol. 258, G878-G886 (1990)
[PubMed]

Iwai, H.

Johnston, C. L.

Y. E. Korchev, J. Gorelik, M. J. Lab, E. V. Sviderskaya, C. L. Johnston, C. R. Coombes, I. Vodyanoy, and C. R. W. Edwards, "Cell volume Measurement using Scanning Ion Conductance Microscopy," Biophys. J. 78, 451-457 (2000)
[CrossRef] [PubMed]

Kemper, B.

P. Langehanenberg, B. Kemper, D. Dirksen, and G. V. Bally, "Autofocusing in digital holographic phase contrast microscopy on pure phase objects for live cell imaging," Appl. Opt. 47, 176-181 (2008)
[CrossRef]

Khatibi, S.

L. Allansson, S. Khatibi, T. Gustavsson, F. Blomstrand, T. Olsson, and E. Hansson, "Single cell volume estimation by three dimensional wide field microscopy applied to astroglial primary cultures," J. Neurosci. Meth. 93,1-11 (1999)
[CrossRef]

Kim, Y. R.

Y. R. Kim and L. Ornstein, "Isovolumetric Sphering of Erythrocytes for more accurate and precise cell volume measurement by flow Cytometry," Cytometry 3, 419-427 (1983)
[CrossRef] [PubMed]

Kneen, M.

J. Farinas, M. Kneen, M. Moore, and Verkman, "Plasma membrane water permeability of cultured cells and epithelia measured by light microscopy with spatial filtering," J. Gen. Physiol. 110, 283-296 (1997)
[CrossRef] [PubMed]

Kolb, A.

Korchev, Y. E.

Y. E. Korchev, J. Gorelik, M. J. Lab, E. V. Sviderskaya, C. L. Johnston, C. R. Coombes, I. Vodyanoy, and C. R. W. Edwards, "Cell volume Measurement using Scanning Ion Conductance Microscopy," Biophys. J. 78, 451-457 (2000)
[CrossRef] [PubMed]

Lab, M. J.

Y. E. Korchev, J. Gorelik, M. J. Lab, E. V. Sviderskaya, C. L. Johnston, C. R. Coombes, I. Vodyanoy, and C. R. W. Edwards, "Cell volume Measurement using Scanning Ion Conductance Microscopy," Biophys. J. 78, 451-457 (2000)
[CrossRef] [PubMed]

Langehanenberg, P.

P. Langehanenberg, B. Kemper, D. Dirksen, and G. V. Bally, "Autofocusing in digital holographic phase contrast microscopy on pure phase objects for live cell imaging," Appl. Opt. 47, 176-181 (2008)
[CrossRef]

Lee, C. M.

C. M. Lee, "Measurement of volume injected into individual cells by quantitative fluorescence microscopy," J. Cell Sci 94, 443-447 (1989)
[PubMed]

Ligthart, G.

F. C. A. Groen, I. T. Young, and G. Ligthart, "A Comparison of Different Focus Functions for Use in Autofocus Algorithms," Cytometry 6, 81-91 (1985)
[CrossRef] [PubMed]

Lue, N.

Lykotrafitis, G.

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, "Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum," PNAS.  105, 13730-13735 (2008)
[CrossRef] [PubMed]

Magistretti, P.

Magistretti, P. J.

Marquet, P.

Michel, B.

Miyamoto, M.

T. Nakahari, M. Murakami, H. Yoshida, M. Miyamoto, Y. Sohma, and Y. Imai, "Decrease in rat submandibular acinar cell volume during Ach stimulation," Am. J. Physiol. 258, G878-G886 (1990)
[PubMed]

Moore, M.

J. Farinas, M. Kneen, M. Moore, and Verkman, "Plasma membrane water permeability of cultured cells and epithelia measured by light microscopy with spatial filtering," J. Gen. Physiol. 110, 283-296 (1997)
[CrossRef] [PubMed]

Murakami, M.

T. Nakahari, M. Murakami, H. Yoshida, M. Miyamoto, Y. Sohma, and Y. Imai, "Decrease in rat submandibular acinar cell volume during Ach stimulation," Am. J. Physiol. 258, G878-G886 (1990)
[PubMed]

Nakahari, T.

T. Nakahari, M. Murakami, H. Yoshida, M. Miyamoto, Y. Sohma, and Y. Imai, "Decrease in rat submandibular acinar cell volume during Ach stimulation," Am. J. Physiol. 258, G878-G886 (1990)
[PubMed]

Nugent, K. A.

C. L. Curl, C. J. Bellair, P. J. Harris, B. E. Allman, A. Roberts, K. A. Nugent, and L. M. D. Delbridge, "Single Cell Volume Measurement by Quantitative Phase Microscopy (QPM): A Case Study of Erythrocyte Morphology," Cell Physiol Biochem 17, 193-200 (2006)
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Y. R. Kim and L. Ornstein, "Isovolumetric Sphering of Erythrocytes for more accurate and precise cell volume measurement by flow Cytometry," Cytometry 3, 419-427 (1983)
[CrossRef] [PubMed]

Park, Y.

Popescu, G.

Rappaz, B.

Roberts, A.

C. L. Curl, C. J. Bellair, P. J. Harris, B. E. Allman, A. Roberts, K. A. Nugent, and L. M. D. Delbridge, "Single Cell Volume Measurement by Quantitative Phase Microscopy (QPM): A Case Study of Erythrocyte Morphology," Cell Physiol Biochem 17, 193-200 (2006)
[CrossRef] [PubMed]

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H. Satoh, L. M. D. Delbridge, L. A. Blatter, and D. M. Bers, "Volume Relationship in Cardiac Myocytes Studied with Confocal Microscopy and Membrane Capacitance Measurements: Species-Dependence and Developmental Effects," Biophys. J. 70, 1494-1504 (1996)
[CrossRef] [PubMed]

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Seung, H. S.

Sohma, Y.

T. Nakahari, M. Murakami, H. Yoshida, M. Miyamoto, Y. Sohma, and Y. Imai, "Decrease in rat submandibular acinar cell volume during Ach stimulation," Am. J. Physiol. 258, G878-G886 (1990)
[PubMed]

Song, S.

Spring, K. R.

P. J. Harris, J. Y. Chatton, P. H. Tran, P. M. Bungay, and K. R. Spring, "pH, morphology, and diffusion in lateral intercellular spaces of epithelial cell monolayers," Am. J. Physiol. 266, C73-C80 (1994)
[PubMed]

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Y. E. Korchev, J. Gorelik, M. J. Lab, E. V. Sviderskaya, C. L. Johnston, C. R. Coombes, I. Vodyanoy, and C. R. W. Edwards, "Cell volume Measurement using Scanning Ion Conductance Microscopy," Biophys. J. 78, 451-457 (2000)
[CrossRef] [PubMed]

Thamm, E.

Tran, P. H.

P. J. Harris, J. Y. Chatton, P. H. Tran, P. M. Bungay, and K. R. Spring, "pH, morphology, and diffusion in lateral intercellular spaces of epithelial cell monolayers," Am. J. Physiol. 266, C73-C80 (1994)
[PubMed]

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Vodyanoy, I.

Y. E. Korchev, J. Gorelik, M. J. Lab, E. V. Sviderskaya, C. L. Johnston, C. R. Coombes, I. Vodyanoy, and C. R. W. Edwards, "Cell volume Measurement using Scanning Ion Conductance Microscopy," Biophys. J. 78, 451-457 (2000)
[CrossRef] [PubMed]

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T. Nakahari, M. Murakami, H. Yoshida, M. Miyamoto, Y. Sohma, and Y. Imai, "Decrease in rat submandibular acinar cell volume during Ach stimulation," Am. J. Physiol. 258, G878-G886 (1990)
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P. J. Harris, J. Y. Chatton, P. H. Tran, P. M. Bungay, and K. R. Spring, "pH, morphology, and diffusion in lateral intercellular spaces of epithelial cell monolayers," Am. J. Physiol. 266, C73-C80 (1994)
[PubMed]

T. Nakahari, M. Murakami, H. Yoshida, M. Miyamoto, Y. Sohma, and Y. Imai, "Decrease in rat submandibular acinar cell volume during Ach stimulation," Am. J. Physiol. 258, G878-G886 (1990)
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[CrossRef] [PubMed]

Y. E. Korchev, J. Gorelik, M. J. Lab, E. V. Sviderskaya, C. L. Johnston, C. R. Coombes, I. Vodyanoy, and C. R. W. Edwards, "Cell volume Measurement using Scanning Ion Conductance Microscopy," Biophys. J. 78, 451-457 (2000)
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[CrossRef] [PubMed]

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Y. R. Kim and L. Ornstein, "Isovolumetric Sphering of Erythrocytes for more accurate and precise cell volume measurement by flow Cytometry," Cytometry 3, 419-427 (1983)
[CrossRef] [PubMed]

F. C. A. Groen, I. T. Young, and G. Ligthart, "A Comparison of Different Focus Functions for Use in Autofocus Algorithms," Cytometry 6, 81-91 (1985)
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L. Allansson, S. Khatibi, T. Gustavsson, F. Blomstrand, T. Olsson, and E. Hansson, "Single cell volume estimation by three dimensional wide field microscopy applied to astroglial primary cultures," J. Neurosci. Meth. 93,1-11 (1999)
[CrossRef]

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Figures (8)

Fig. 1.
Fig. 1.

Schematic diagram of our experimental setup for QPM.

Fig. 2.
Fig. 2.

(a) Measured interferogram images of a polystyrene bead for 5 different object positions from -2 μm to +2 μm with the step size of 1 μm. (b) The reconstructed amplitude images from the interferograms displayed in Fig. 2(a). (c) Calculated focus measures of SGA and LFA for the five reconstructed amplitude images obtained in Fig. 2(a). Each graph is normalized by its peak value.

Fig. 3.
Fig. 3.

(a) Calculate phase images of the polystyrene bead obtained from the interferograms displayed in Fig. 2(a). (b) The thickness line profiles of the sample indicated by each arrow in Fig. 3(a). (c) The thickness line profile of the sample at the focus position (z = 0) and the simulated thickness profile for a perfect sphere.

Fig. 4.
Fig. 4.

Measured volume of a polystyrene bead with respect to focus position.

Fig. 5.
Fig. 5.

Optical path length fluctuation in a series of phase images without a sample.

Fig. 6.
Fig. 6.

(a) and (b) Quantitative phase images of a single RBC. (c) and (d) Thickness line profiles along the vertical and the horizontal arrows indicated in Fig. 6(a).

Fig. 7.
Fig. 7.

(a) Calculated boundary line of an RBC calculated from the phase image of Fig. 6(a). (b) The dilated binary boundary image modified from Fig. 7(a). (c) Center-filled binary mask modified from Fig. 7(b). (d) Background suppressed phase image by multiplying the binary mask of Fig. 7(c) to the original phase image in Fig. 6(a).

Fig. 8.
Fig. 8.

Measured volumes of a single RBC with and without our background suppression method at a measurement speed of 100 frames per second.

Equations (6)

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I ( x , y ) = I R + I S ( x , y ) + 2 I R · I S ( x , y ) cos [ q x x + q y y + ϕ ( x , y ) ] ,
q max = 1 λ sin ( θ max )
SGA [ ( f ( x , y ) x ) 2 + ( f ( x , y ) y ) 2 ] dxdy
LFA [ ( 2 f ( x , y ) x 2 ) + ( 2 f ( x , y ) y 2 ) ] 2 dxdy
ϕ ( x , y ) = 2 π λ Δ n · t ( x , y ) ,
g ( x , y ) ϕ 1 ( x , y ) 2

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