Prediction of sperm progression in three dimensions using rapid optical imaging and dynamic mechanical modeling

Abstract We present a multidisciplinary approach for predicting how sperm cells with various morphologies swim in three-dimensions (3D), over time scales of milliseconds to hours at spatial resolutions of less than half a micron. We created the sperm 3D geometry and built a numerical mechanical model using the experimentally acquired dynamic 3D refractive index profiles of sperm cells swimming freely in vitro as imaged by high-resolution optical diffraction tomography. By controlling parameters in the model, such as the size and shape of the sperm head and tail, we can then predict how different sperm cells, normal or abnormal, would swim in 3D, in the short or long term. We quantified various 3D structural factor effects on the sperm long-term motility. We found that some abnormal sperm cells swim faster than normal sperm cells, in contrast to the commonly-used sperm selection assumption during IVF, according to which sperm cells should mainly be chosen based on their progressive motion. We established a new tool for sperm analysis and male-infertility diagnosis, as well as new sperm selection criteria for fertility treatments.

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Label-free monitoring of 3D cortical neuronal growth in vitro using optical diffraction tomography

10.1101/2021.07.31.454602 ◽

2021 ◽

Author(s):

Ariel J. Lee ◽

DongJo Yoon ◽

SeungYun Han ◽

Herve Hugonnet ◽

WeiSun Park ◽

...

Keyword(s):

Optical Diffraction ◽

Neuronal Cultures ◽

Multiple Time ◽

Diffraction Tomography ◽

Label Free ◽

Neuronal Growth ◽

Monitoring Method ◽

Optical Diffraction Tomography ◽

Immunofluorescence Labeling

The highly complex central nervous systems of mammals are often studied using three-dimensional (3D) in vitro primary neuronal cultures. A coupled confocal microscopy and immunofluorescence labeling are widely utilized for visualizing the 3D structures of neurons. However, this requires fixation of the neurons and is not suitable for monitoring an identical sample at multiple time points. Thus, we propose a label-free monitoring method for 3D neuronal growth based on refractive index tomograms obtained by optical diffraction tomography. The 3D morphology of the neurons was clearly visualized, and the developmental processes of neurite outgrowth in 3D spaces were analyzed for individual neurons.

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286 PRODUCTION OF NORMAL MICE USING LONG-TERM PRESERVED MOUSE SPERMATOZOA WITHOUT FREEZING

Reproduction Fertility and Development ◽

10.1071/rdv23n1ab286 ◽

2011 ◽

Vol 23 (1) ◽

pp. 241

Author(s):

C. Li ◽

E. Mizutani ◽

T. Ono ◽

T. Wakayama

Keyword(s):

Low Cost ◽

Blastocyst Stage ◽

Isolation Medium ◽

Safety Issues ◽

Fertility Treatments ◽

Preservation Method ◽

Refrigerator Temperature ◽

Nuclear Isolation

Mammalian spermatozoa preservation now plays an important role in fertility treatments, generating hybrid animals and protecting endangered and extinct species. To date, the most common method of sperm preservation is freezing in liquid nitrogen (LN2). However, this method requires constant supplementation of LN2 and also presents some safety issues involved in transporting LN2. Here we describe a new sperm preservation method that does not involve freezing. Mouse spermatozoa were cultured in four basic media (HEPES–Chatot-Ziomele-Barister’s medium (HCZB), KSOM, K+-rich nuclear isolation medium (NIM), and PBS) with or without 10% BSA or 15% Ficoll as a cryoprotectant, and preserved in a refrigerator for up to 6 months. These preserved sperm were then injected into fresh oocytes and cultured to the blastocyst stage in vitro or transferred into recipient females to demonstrate their genetic integrity. Oocytes injected with 1-month-preserved spermatozoa in NIM and PBS showed significantly higher blastocyst rates (22.8% and 18.9%) than those in HEPES-CZB and KSOM (1-way ANOVA, P < 0.05). In embryos with 3-month-preserved spermatozoa in NIM or PBS with BSA or Ficoll, 5.3–24.0%; P < 0.05 of embryos, (n = 1056) developed to the blastocyst stage, and the developmental ratio was not decreased even for 6-month preservation (13.6–18.2%; P > 0.05). Surprisingly, 18 pups were obtained using spermatozoa stored in those mediums for 6 months. Moreover, this new method allowed easy production of healthy offspring even after transporting spermatozoa between two countries by aircraft at room temperature without any protection. In conclusion, this method allows for easy long-term preservation of mouse spermatozoa in a simple, modified medium at refrigerator temperature with very low cost and wide application.

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Label-free monitoring of 3D cortical neuronal growth in vitro using optical diffraction tomography

Biomedical Optics Express ◽

10.1364/boe.439404 ◽

2021 ◽

Author(s):

Ariel Lee ◽

DongJo Yoon ◽

SeungYun Han ◽

Herve Hugonnet ◽

Wei Sun Park ◽

...

Keyword(s):

Optical Diffraction ◽

Diffraction Tomography ◽

Label Free ◽

Neuronal Growth ◽

Optical Diffraction Tomography

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Label-free three-dimensional imaging and quantitative analysis of long-term on-chip vasculogenesis using optical diffraction tomography

Quantitative Phase Imaging VII ◽

10.1117/12.2584889 ◽

2021 ◽

Author(s):

Chungha Lee ◽

Seunggyu Kim ◽

Herve Hugonnet ◽

Jessie S. Jeon ◽

WeiSun Park ◽

...

Keyword(s):

Quantitative Analysis ◽

Three Dimensional ◽

Optical Diffraction ◽

Diffraction Tomography ◽

Label Free ◽

Three Dimensional Imaging ◽

Optical Diffraction Tomography ◽

On Chip

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Measurements of morphological and biochemical alterations in individual neuron cells associated with early neurotoxic effects in Parkinson’s disease

10.1101/080937 ◽

2016 ◽

Cited By ~ 3

Author(s):

Su-A Yang ◽

Jonghee Yoon ◽

Kyoohyun Kim ◽

YongKeun Park

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Optical Diffraction ◽

Optical Measurements ◽

Diffraction Tomography ◽

Biochemical Alterations ◽

Optical Diffraction Tomography ◽

Neurotoxic Effects ◽

Index Distribution

AbstractParkinson’s disease (PD) is a common neurodegenerative disease. However, therapeutic methods of PD are still limited due to complex pathophysiology in PD. Here, we present optical measurements of individual neurons fromin vitroPD model using optical diffraction tomography (ODT). By measuring 3-D refractive index distribution of neurons, morphological and biochemical alterations inin-vitroPD model are quantitatively investigated. We found that neurons show apoptotic features in early PD progression. The present approach will open up new opportunities for quantitative investigation of the pathophysiology of various neurodegenerative diseases.

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Intracellular Detection and Localization of Nanoparticles by Refractive Index Measurement

Sensors ◽

10.3390/s21155001 ◽

2021 ◽

Vol 21 (15) ◽

pp. 5001

Author(s):

Alain Géloën ◽

Karyna Isaieva ◽

Mykola Isaiev ◽

Olga Levinson ◽

Emmanuelle Berger ◽

...

Keyword(s):

Refractive Index ◽

Optical Diffraction ◽

Live Cells ◽

Diffraction Tomography ◽

Refractive Index Measurement ◽

Optical Diffraction Tomography ◽

Fluorescence Measurements ◽

Index Measurement ◽

Intracellular Detection

The measuring of nanoparticle toxicity faces an important limitation since it is based on metrics exposure, the concentration at which cells are exposed instead the true concentration inside the cells. In vitro studies of nanomaterials would benefit from the direct measuring of the true intracellular dose of nanoparticles. The objective of the present study was to state whether the intracellular detection of nanodiamonds is possible by measuring the refractive index. Based on optical diffraction tomography of treated live cells, the results show that unlabeled nanoparticles can be detected and localized inside cells. The results were confirmed by fluorescence measurements. Optical diffraction tomography paves the way to measuring the true intracellular concentrations and the localization of nanoparticles which will improve the dose-response paradigm of pharmacology and toxicology in the field of nanomaterials.

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Partially Coherent Optical Diffraction Tomography Toward Practical Cell Study

Frontiers in Physics ◽

10.3389/fphy.2021.666256 ◽

2021 ◽

Vol 9 ◽

Author(s):

Juan M. Soto ◽

José A. Rodrigo ◽

Tatiana Alieva

Keyword(s):

Refractive Index ◽

Computational Imaging ◽

Optical Diffraction ◽

Wide Field ◽

Diffraction Tomography ◽

Diatom Frustule ◽

African Green Monkey Kidney ◽

Partially Coherent ◽

Optical Diffraction Tomography

Optical diffraction tomography (ODT) is a computational imaging technique based on refractive index (RI) contrast. Its application for microscopic imaging of weakly absorbing and scattering samples has been demonstrated by using a specially designed holographic microscope with angular scanning of the coherent sample illumination direction. Recently, an alternative low cost technique based on partially coherent sample illumination (PC-ODT), which is compatible with the conventional wide-field transmission microscope, has been established. In this case, the 3D refractive index distribution of the sample is obtained by deconvolution from a single stack of through-focus intensity images. The performance of PC-ODT has been successfully tested on various fixed specimens (diatom frustule and biological cells) and moving bacteria. Here, we demonstrate that the PC-ODT is an efficient tool for the analysis of living eukaryotic cell dynamics at short- and long-term periods. The COS-7 cells, which hail from the African green monkey kidney, have been chosen for this study. A fast data acquisition setup comprising an optical scanning module can be easily attached to the microscope, and it allows observing cell 3D organelle movements and RI variations, with the required temporal resolution. In particular, a more rapid nucleoli rotation than previously reported has been found. The long-term cell monitoring during necrosis reveals significant changes in cell dry mass concentration obtained from recovered RI contrast.

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Fertility Status and Cancer

Seminars in Reproductive Medicine ◽

10.1055/s-0037-1603098 ◽

2017 ◽

Vol 35 (03) ◽

pp. 291-297 ◽

Cited By ~ 1

Author(s):

Louise Brinton

Keyword(s):

Cancer Risk ◽

Small Sample ◽

Ovarian Cancer Risk ◽

Long Term Effects ◽

Vitro Fertilization ◽

Fertility Treatments ◽

Nulliparous Women ◽

Small Sample Sizes

AbstractElevated estradiol levels associated with ovulation-stimulating drugs used to treat infertility and ovarian punctures involved with in vitro fertilization (IVF) have led to concerns regarding long-term cancer effects. Assessed risks, however, have been contradictory. This may reflect small sample sizes and short follow-up times, as well as an inability of many studies to adjust for appropriate potential confounders, such as causes of infertility and parity. While early studies of ovulation-stimulating drugs suggested large increases in large ovarian cancer risk, more recent studies have not confirmed this. However, several investigations have noted risk elevations associated with drug use or IVF among women who remain nulliparous, possibly due to resistant infertility. Studies also suggest increases in borderline ovarian tumors, although the effects of surveillance bias are unclear. Investigations have not shown large increases in breast cancer risk, although a few studies suggest possible increases in select subgroups, including nulliparous women or those with high drug dosages in the distant past. A few studies have evaluated risks of other cancers (endometrium, colorectum, thyroid, and melanoma), without definitive results. Since women exposed to fertility treatments are just beginning to enter typical cancer age ranges, further well-designed investigations are needed to fully delineate long-term effects.

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Ultrastructural investigations of MDL 19,660-induced effects on the canine platelet and megakaryocyte

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100159412 ◽

1990 ◽

Vol 48 (3) ◽

pp. 374-375

Author(s):

D.E. Loudy ◽

J. Sprinkle-Cavallo ◽

J.T. Yarrington ◽

F.Y. Thompson ◽

J.P. Gibson

Keyword(s):

Antidepressant Drug ◽

Beagle Dogs ◽

Long Term Effects ◽

Short Term ◽

Platelet Counts ◽

Toxicological Studies ◽

Induced Aggregation ◽

Internal Architecture

Previous short term toxicological studies of one to two weeks duration have demonstrated that MDL 19,660 (5-(4-chlorophenyl)-2,4-dihydro-2,4-dimethyl-3Hl, 2,4-triazole-3-thione), an antidepressant drug, causes a dose-related thrombocytopenia in dogs. Platelet counts started to decline after two days of dosing with 30 mg/kg/day and continued to decrease to their lowest levels by 5-7 days. The loss in platelets was primarily of the small discoid subpopulation. In vitro studies have also indicated that MDL 19,660: does not spontaneously aggregate canine platelets and has moderate antiaggregating properties by inhibiting ADP-induced aggregation. The objectives of the present investigation of MDL 19,660 were to evaluate ultrastructurally long term effects on platelet internal architecture and changes in subpopulations of platelets and megakaryocytes.Nine male and nine female beagle dogs were divided equally into three groups and were administered orally 0, 15, or 30 mg/kg/day of MDL 19,660 for three months. Compared to a control platelet range of 353,000- 452,000/μl, a doserelated thrombocytopenia reached a maximum severity of an average of 135,000/μl for the 15 mg/kg/day dogs after two weeks and 81,000/μl for the 30 mg/kg/day dogs after one week.

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