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Oocyte/embryo vitrification is one of the basic techniques employed in assisted reproduction and fertility preservation. However, the current process typically requires manual operation by experienced embryologists, which is both time-consuming and exhibits inconsistent outcomes. To resolve this issue, we herein developed an automated vitrification device for oocytes/embryos. The device consists of a microfluidic mixing unit, a microgrid capillary, and a mechanical sliding unit. The microfluidic mixing unit was adopted to determine continuous changes in cryoprotective agent (CPA) concentration, reducing osmotic damage during CPA loading/removal; and the microgrid capillary was used to load/remove the CPA and vitrify the oocytes/embryos in the same carrier so as to reduce cellular loss due to cell transfer. The medical absorbent dressing was placed under the carrier, and the excess liquid was absorbed to minimize the remaining CPA solution after CPA loading. Eight different loading/removal curves were developed for CPA loading and removal protocols, and we conducted oocyte vitrification with automated vitrification equipment. Our results revealed that a quadratic function-based, loading-removal curve achieved the highest oocyte survival rate within an 8-min loading and removal duration. In addition, there was no significant difference in oocyte survival rates between the automated vitrification device and the Cryotop multi-step equilibration method (90.33 % and 94.33 %, respectively); nor did the two methods differ in terms of survival or hatching rates of 8-cell embryos. The current system automates and standardizes the oocyte/embryo vitrification process while achieving survival and developmental potential.
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http://dx.doi.org/10.1016/j.cryobiol.2025.105275 | DOI Listing |
Device
June 2025
Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Boston, MA, USA.
Oocyte cryopreservation is a critical step of fertilization (IVF). The handling and loading of cryoprotectant currently rely on manual pipetting methods, which leads to outcome variations, high cost and limited accessibility. Here we developed a low-cost and user-friendly microfluidic device to automate the complex multistep process of oocyte CPA loading, delivering cryopreservation-ready oocytes.
View Article and Find Full Text PDFFront Cell Dev Biol
July 2025
Guangdong Second Provincial General Hospital, Guangzhou, China.
[This corrects the article DOI: 10.3389/fcell.2023.
View Article and Find Full Text PDFbioRxiv
May 2025
School of Chemical, Biological and Environmental Engineering, Oregon State University.
Vitrification is a promising approach for cryopreserving complex biological structures such as organs. However, to prevent ice formation, high concentrations of cell-permeable cryoprotective agents (CPAs) are required, which can be highly toxic. The current reliance on a small number of CPAs limits optimization of low-toxicity compositions for vitrification.
View Article and Find Full Text PDFCurr Opin Struct Biol
August 2025
State Key Laboratory of Membrane Biology, Peking-Tsinghua Center for Life Sciences, School of Life Sciences, Peking University, Beijing 100871, China; Changping Laboratory, Beijing 102206, China. Electronic address:
Cryo-electron tomography (cryo-ET) is revolutionizing in situ structural analysis of single-cell specimens, yet its application to tissues has been hindered, primarily due to challenges posed by tissue thickness. Advances in sample vitrification, cryo-focused ion beam (cryo-FIB) milling, and lift-out techniques have substantially improved tissue preparation, enabling thin, electron microscopy-compatible samples. Furthermore, the integration of automation, complementary imaging modalities, and AI has streamlined imaging workflows and data analysis.
View Article and Find Full Text PDFCryobiology
June 2025
Institute of Biothermal Science & Technology, University of Shanghai for Science and Technology, Shanghai, 200093, China; Shanghai Collaborative Innovation Center for Tumor Energy Therapy, Shanghai, 200093, China; Shanghai Technical Service Platform for Cryopreservation of Biological Resources, Shan
Oocyte/embryo vitrification is one of the basic techniques employed in assisted reproduction and fertility preservation. However, the current process typically requires manual operation by experienced embryologists, which is both time-consuming and exhibits inconsistent outcomes. To resolve this issue, we herein developed an automated vitrification device for oocytes/embryos.
View Article and Find Full Text PDF