Probing red blood cell morphology using high-frequency photoacoustics.

A method that can rapidly quantify variations in the morphology of single red blood cells (RBCs) using light and sound is presented. When irradiated with a laser pulse, an RBC absorbs the optical energy and emits an ultrasonic pressure wave called a photoacoustic wave. The power spectrum of the resulting photoacoustic wave contains distinctive features that can be used to identify the RBC size and morphology. When particles 5-10 μm in diameter (such as RBCs) are probed with high-frequency photoacoustics, unique periodically varying minima and maxima occur throughout the photoacoustic signal power spectrum at frequencies >100 MHz. The location and distance between spectral minima scale with the size and morphology of the RBC; these shifts can be used to quantify small changes in the morphology of RBCs. Morphological deviations from the normal biconcave RBC shape are commonly associated with disease or infection. Using a single wide-bandwidth transducer sensitive to frequencies between 100 and 500 MHz, we were able to differentiate healthy RBCs from irregularly shaped RBCs (such as echinocytes, spherocytes, and swollen RBCs) with high confidence using a sample size of just 21 RBCs. As each measurement takes only seconds, these methods could eventually be translated to an automated device for rapid characterization of RBC morphology and deployed in a clinical setting to help diagnose RBC pathology.

[1]  Lihong V. Wang,et al.  Subwavelength-resolution label-free photoacoustic microscopy of optical absorption in vivo. , 2010, Optics letters.

[2]  Lihong V. Wang,et al.  Noninvasive laser-induced photoacoustic tomography for structural and functional in vivo imaging of the brain , 2003, Nature Biotechnology.

[3]  P. Canham.  The minimum energy of bending as a possible explanation of the biconcave shape of the human red blood cell. , 1970, Journal of theoretical biology.

[4]  J W Hunt,et al.  High-frequency ultrasound scattering from microspheres and single cells. , 2005, The Journal of the Acoustical Society of America.

[5]  S. Chien,et al.  Red cell rheology in stomatocyte-echinocyte transformation: roles of cell geometry and cell shape. , 1986, Blood.

[6]  R. Lemor,et al.  Developing a high-resolution photoacoustic microscopy platform , 2009 .

[7]  Lihong V. Wang,et al.  Photoacoustic Tomography: In Vivo Imaging from Organelles to Organs , 2012, Science.

[8]  Roy G. M. Kolkman,et al.  In vivo photoacoustic imaging of blood vessels using an extreme-narrow aperture sensor , 2003 .

[9]  V. Kachel,et al.  Uniform lateral orientation, caused by flow forces, of flat particles in flow-through systems. , 1977, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[10]  Qifa Zhou,et al.  Optimal ultraviolet wavelength for in vivo photoacoustic imaging of cell nuclei. , 2012, Journal of biomedical optics.

[11]  W. Grill,et al.  Age-Dependent Acoustic and Microelastic Properties of Red Blood Cells Determined by Vector Contrast Acoustic Microscopy , 2012, Microscopy and Microanalysis.

[12]  N. Gov,et al.  Red blood cell membrane fluctuations and shape controlled by ATP-induced cytoskeletal defects. , 2005, Biophysical journal.

[13]  Y. C. Fung,et al.  Improved measurements of the erythrocyte geometry. , 1972, Microvascular research.

[14]  Subhajit Karmakar,et al.  Computational Investigation on the Photoacoustics of Malaria Infected Red Blood Cells , 2012, PloS one.

[15]  Michael C. Kolios,et al.  Photoacoustic ultrasound spectroscopy for assessing red blood cell aggregation and oxygenation , 2012, Journal of biomedical optics.

[16]  L. Zolla,et al.  Red blood cell storage and cell morphology , 2012, Transfusion medicine.

[17]  Subra Suresh,et al.  Measuring single-cell density , 2011, Proceedings of the National Academy of Sciences.

[18]  Frank Stracke,et al.  Gigahertz optoacoustic imaging for cellular imaging , 2010, BiOS.

[19]  Lihong V. Wang,et al.  Functional photoacoustic microscopy for high-resolution and noninvasive in vivo imaging , 2006, Nature Biotechnology.

[20]  A. C. Burton,et al.  Distribution of Size and Shape in Populations of Normal Human Red Cells , 1968, Circulation research.

[21]  Yongkeun Park,et al.  Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum , 2008, Proceedings of the National Academy of Sciences.

[22]  K Zouaoui Boudjeltia,et al.  Assessment of erythrocyte shape by flow cytometry techniques , 2006, Journal of Clinical Pathology.

[23]  Lihong V. Wang,et al.  Photoacoustic imaging in biomedicine , 2006 .

[24]  Hao Zhang,et al.  Imaging of hemoglobin oxygen saturation variations in single vessels in vivo using photoacoustic microscopy , 2007 .

[25]  David J. Clarke,et al.  Cell manipulation in ultrasonic standing wave fields , 2007 .

[26]  P. J. Goetz,et al.  Ultrasonic characterization of proteins and blood cells. , 2006, Colloids and surfaces. B, Biointerfaces.

[27]  John M. Reid,et al.  Scattering of Ultrasound by Blood , 1976, IEEE Transactions on Biomedical Engineering.

[28]  Lihong V. Wang,et al.  Optical-resolution photoacoustic microscopy for in vivo imaging of single capillaries. , 2008, Optics letters.

[29]  G Cloutier,et al.  A system-based approach to modeling the ultrasound signal backscattered by red blood cells. , 1999, Biophysical journal.

[30]  J. Acker,et al.  Biopreservation of red blood cells: past, present, and future. , 2005, Transfusion medicine reviews.

[31]  Lihong V. Wang.  Multiscale photoacoustic microscopy and computed tomography. , 2009, Nature photonics.

[32]  Paulette Mehta,et al.  Wintrobe’s Clinical Hematology , 2009 .

[33]  G Cloutier,et al.  Ultrasound backscattering from non-aggregating and aggregating erythrocytes--a review. , 1997, Biorheology.

[34]  Michael C. Kolios,et al.  On the use of photoacoustics to detect red blood cell aggregation , 2012, Biomedical optics express.

[35]  M. I. Khan,et al.  Photoacoustic "Signatures" of Particulate Matter: Optical Production of Acoustic Monopole Radiation , 1990, Science.

[36]  Eric M. Strohm,et al.  Photoacoustic measurements of single red blood cells , 2012, 2012 IEEE International Ultrasonics Symposium.

[37]  P. Beard.  Biomedical photoacoustic imaging , 2011, Interface Focus.

[38]  M. Socol,et al.  Full dynamics of a red blood cell in shear flow , 2012, Proceedings of the National Academy of Sciences.

[39]  Ivan Gorelikov,et al.  Vaporization of perfluorocarbon droplets using optical irradiation , 2011, Biomedical optics express.

[40]  S. Chien.  Red cell deformability and its relevance to blood flow. , 1987, Annual review of physiology.

[41]  J. Hudson,et al.  Flow cytometric analysis of human erythrocytes: I. Probed with lectins and immunoglobulins , 1991, Experimental Gerontology.

[42]  Scott R. Manalis,et al.  Measuring the mass, density, and size of particles and cells using a suspended microchannel resonator , 2007 .

[43]  Michael C. Kolios,et al.  Quantitative measurements of apoptotic cell properties using acoustic microscopy , 2010, IEEE Transactions on Ultrasonics, Ferroelectrics and Frequency Control.

[44]  Ivan Gorelikov,et al.  Acoustic and photoacoustic characterization of micron-sized perfluorocarbon emulsions , 2012, Journal of biomedical optics.

[45]  M. Bessis,et al.  Present status of spiculed red cells and their relationship to the discocyte-echinocyte transformation: a critical review. , 1972, Blood.

[46]  Geng Ku,et al.  Photoacoustic microscopy with 2-microm transverse resolution. , 2010, Journal of biomedical optics.