Strategies for locating the female gamete: the importance of measuring sperm trajectories in three spatial dimensions

Guerrero, Adán; Carneiro, Jorge; Pimentel, Arturo; Wood, Christopher D.; Corkidi, Gabriel; Darszon, Alberto

doi:10.1093/molehr/gar042

Cited by 53 publications

(41 citation statements)

References 113 publications

(225 reference statements)

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“…Conventional microscopes equipped with high-magnification objective lenses and high-frame-rate cameras can only meet these requirements for imaging sperms along a 2D plane, which can infer limited information on their natural 3D motion (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16). Estimation of the 3D trajectories of sperms from their 2D observations can also be feasible in some cases by assuming a known swimming pattern (15,29). However, such approaches in general would not be able to infer the details and quantify the fine parameters of 3D sperm trajectories due to lack of position information along the third dimension.…”

Section: Discussionmentioning

confidence: 99%

“…Second, when swimming near a surface, sea urchin sperms tend to follow circular swimming paths with a strongly preferred handedness (9,11,31,34,35), whereas human sperms do not exhibit such behavior. Third, helical trajectories of human sperms can be observed both in free 3D volume and near solid surfaces; however, sea urchin sperms only display helical movement in free 3D volume (17)(18)(19)29). Fourth, the helical trajectories of human sperms, compared to sea urchin sperms (19), exhibit significantly smaller helix radii (1.6 AE 0.5 μm vs. 6.8 AE 1.1 μm) and faster rotation speeds (6.8 AE 4.6 r∕s vs. 4.0 AE 0.8 r∕s), making them much more challenging to resolve in 3D.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

High-throughput lensfree 3D tracking of human sperms reveals rare statistics of helical trajectories

Ozcan

2012

Proc. Natl. Acad. Sci. U.S.A.

314

277

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Dynamic tracking of human sperms across a large volume is a challenging task. To provide a high-throughput solution to this important need, here we describe a lensfree on-chip imaging technique that can track the three-dimensional (3D) trajectories of >1,500 individual human sperms within an observation volume of approximately 8-17 mm 3 . This computational imaging platform relies on holographic lensfree shadows of sperms that are simultaneously acquired at two different wavelengths, emanating from two partially-coherent sources that are placed at 45°with respect to each other. This multiangle and multicolor illumination scheme permits us to dynamically track the 3D motion of human sperms across a field-of-view of >17 mm 2 and depth-of-field of approximately 0.5-1 mm with submicron positioning accuracy. The large statistics provided by this lensfree imaging platform revealed that only approximately 4-5% of the motile human sperms swim along well-defined helices and that this percentage can be significantly suppressed under seminal plasma. Furthermore, among these observed helical human sperms, a significant majority (approximately 90%) preferred right-handed helices over left-handed ones, with a helix radius of approximately 0.5-3 μm, a helical rotation speed of approximately 3-20 rotations∕s and a linear speed of approximately 20-100 μm∕s. This high-throughput 3D imaging platform could in general be quite valuable for observing the statistical swimming patterns of various other microorganisms, leading to new insights in their 3D motion and the underlying biophysics.human sperm imaging | sperm tracking | digital holography | micro-swimmer O bserving three-dimensional (3D) trajectories of sperms is in general a challenging task. This is partially due to limited imaging volume of optical microscopes that are based on conventional lenses. For human sperms this becomes even more challenging since the sperm head is small (approximately 3-4 μm) demanding a relatively high-magnification objective lens, and moves rather fast (20-100 μm∕s) which makes it difficult to track their 3D swimming patterns as they quickly move out of the observation volume of an objective lens. Partly due to this low throughput and the limited spatial and temporal sampling windows that conventional microscopes provide, natural 3D swimming patterns of human sperms and their statistics could not be reported so far. Earlier results (1-19) that were obtained using lens-based conventional microscopes either measured the 2D trajectories of the human sperms along a focal plane, or reported on sperms of other species such as sea urchin, which were significantly easier to resolve under a microscope since their 3D rotation diameter is larger (>13 μm) together with a lower rotation frequency compared to human sperms.Here we report a new technique that is based on lensfree holographic imaging on a chip to dynamically track the 3D trajectories of human sperms across a large volume of approximately 8-17 mm 3 ( Fig. 1) with submicron positioning accuracy. Thi...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

High-throughput lensfree 3D tracking of human sperms reveals rare statistics of helical trajectories

Ozcan

2012

Proc. Natl. Acad. Sci. U.S.A.

314

277

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“…Movement results by repetitive cycles of fl agellar bending, arising from microtubule sliding using the force generated by dynein ATPases whose activity is modulated by pH, ATP, ADP, Ca 2+ , and phosphorylation (Christen et al 1983 ;Lindemann and Goltz 1988 ). Ion transport that supports and controls fl agellar beating plays key roles in sperm motility regulation (Guerrero et al 2011 ;Kaupp et al 2008 ).…”

Section: Motilitymentioning

confidence: 99%

Cl− Channels and Transporters in Sperm Physiology

Treviño

Orta

Figueiras-Fierro

et al. 2014

Sexual Reproduction in Animals and Plants

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Spermatozoa must decode environmental and cellular cues to succeed in fertilization, and this process relies heavily on ion channels. New observations bring to light the relevant participation of Cl − channels and anion transporters in some of the main sperm functions. Here we review the evidence that indicates the participation of Cl − channels in motility, maturation, and the acrosome reaction (AR), and what is known about their molecular identity and regulation. Our better understanding of sperm anion transport will yield tools to handle some infertility problems, improve animal breeding and preserve biodiversity, and develop selective and secure male contraceptives.

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“…El intercambio de información entre los gametos, y entre ellos y su entorno, está cifrado en buena medida por cambios en la permeabilidad iónica de sus membranas. Los canales y transportadores iónicos tienen un papel importante en la movilidad ya que de ellos depende el [Ca 2+ ]i que a su vez regula el nado del espermatozoide [1]. Por la complejidad de análisis del movimiento de los flagelos, los estudios reportados han limitado este análisis a dos dimensiones [2,3,4], cuando en la realidad, su movimiento se realiza en un espacio tridimensional.…”

Section: Introductionunclassified

Determinación del error en la medición de fluorescencia 2D del flagelo de espermatozoide humano durante su batido 3D

Montoya¹

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Resumen--La movilidad es una propiedad fundamental de muchas células. El espermatozoide es una de estas células, su flagelo le permite nadar y cumplir con su objetivo principal, fecundar al óvulo. En este sentido, el estudio de cómo el calcio intracelular ( I. INTRODUCCIÓNEn el movimiento del espermatozoide, el flagelo juega un papel fundamental, por lo que entender su funcionamiento es importante para la reproducción que impacta no solo a la fisiología y la salud humana, sino a la ganadería, la pesca y a la preservación de las especies. La reproducción sexual requiere de la comunicación entre los gametos, el espermatozoide y el óvulo, sin embargo los mecanismos moleculares involucrados no se conocen con detalle. El intercambio de información entre los gametos, y entre ellos y su entorno, está cifrado en buena medida por cambios en la permeabilidad iónica de sus membranas. Los canales y transportadores iónicos tienen un papel importante en la movilidad ya que de ellos depende el [Ca 2+ ]i que a su vez regula el nado del espermatozoide [1]. Por la complejidad de análisis del movimiento de los flagelos, los estudios reportados han limitado este análisis a dos dimensiones [2,3,4], cuando en la realidad, su movimiento se realiza en un espacio tridimensional.En diferentes casos, este tipo de estudios 2D usan fluoróforos que son sensibles a los cambios en el [Ca 2+ ]i. Un fluoróforo es una molécula que emite luz al ser excitada, con la particularidad de que las longitudes de onda de absorción y de emisión son diferentes, lo que facilita filtrar la luz para obtener altos contrastes en las imágenes adquiridas. En particular, para este trabajo, se empleó Fluo4-AM, un fluoróforo que se une a las moléculas de calcio, lo cual produce un aumento en la fluorescencia y además es permeable en la membrana celular de los espermatozoides. Con estas características, el Fluo4-AM reporta los niveles de [Ca 2+ ]i presente en el flagelo, siendo la intensidad de fluorescencia el observable experimental.El desenfoque es uno de los problemas que aparecen cuando se pretende medir la intensidad de la fluorescencia proveniente del flagelo (estructura tridimensional) en un solo plano focal a una altura fija. Este efecto se produce por el movimiento 3D de la célula, teniendo como consecuencia que la intensidad de luz emitida por el Fluo4-AM se vea amortiguada en las regiones desenfocadas, llevando a una medida incierta de la concentración de [Ca 2+ ]i.El objetivo de este trabajo es determinar la magnitud del error que se produce al medir en un solo plano óptico (2D) la totalidad de fluorescencia emitida por la estructura tridimensional del flagelo de espermatozoide de humano. Para lograrlo, se utilizó la configuración del sistema reportado en Corkidi et al 2008 [5], en donde se hace oscilar el objetivo de un microscopio para obtener en intervalos de tiempos cortos, diferentes planos focales que contienen la estructura 3D del flagelo, y de esta manera, realizar medidas 3D que minimicen las consecuencias del desenfoque. II. METODOLOGÍAE...

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Strategies for locating the female gamete: the importance of measuring sperm trajectories in three spatial dimensions

Cited by 53 publications

References 113 publications

High-throughput lensfree 3D tracking of human sperms reveals rare statistics of helical trajectories

High-throughput lensfree 3D tracking of human sperms reveals rare statistics of helical trajectories

Cl− Channels and Transporters in Sperm Physiology

Determinación del error en la medición de fluorescencia 2D del flagelo de espermatozoide humano durante su batido 3D

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