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To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
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Automated device for rapid sample cooling via controlled submersion.

Purva Joshi1, Zachary Chau1, Shaun Keating2

  • 1Center for Engineering in Medicine & Surgery, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and Shriners Children's Boston, USA.

Cryobiology
|May 11, 2025
PubMed
Summary
This summary is machine-generated.

A new low-cost, automated device precisely submerges specimens in liquid nitrogen for rapid cooling, crucial for vitrification. This portable system ensures consistent cooling rates comparable to manual methods, enhancing cryopreservation reliability.

Keywords:
Rapid submersionUltra-fast coolingVitrification

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Area of Science:

  • Cryobiology
  • Biotechnology
  • Robotics

Background:

  • Vitrification requires precise, rapid cooling of biological specimens.
  • Current methods for achieving optimal cooling rates can be inconsistent or labor-intensive.
  • Automated solutions are needed to improve the reliability and reproducibility of cryopreservation.

Purpose of the Study:

  • To develop and validate a low-cost, portable automated device for programmable specimen submersion in liquid nitrogen.
  • To enable precise control over cooling rates for vitrification applications.
  • To assess the device's precision, reliability, and practical functionality.

Main Methods:

  • A device utilizing a stepper motor, linear actuator, and 3D-printed robotic arm was designed for programmable vertical sample submersion.
  • The device's precision in positioning was validated by measuring starting height and submersion distance errors.
  • Cooling rates were compared between automated and manual submersion and assessed using a standard insemination straw.

Main Results:

  • The automated device demonstrated high precision, with less than 1.5% relative error in positioning.
  • Automated submersion achieved cooling rates statistically indistinguishable from manual methods.
  • Practical application testing with a 0.25 mL insemination straw confirmed functionality and achieved cooling rates consistent with literature and computational predictions.

Conclusions:

  • The developed automated device offers a precise, reliable, and cost-effective solution for rapid specimen cooling in vitrification.
  • Its portability and compatibility with various cryogenic containers enhance its utility in cryobiology research and applications.
  • The device's performance validates its potential to improve the consistency and success rates of cryopreservation.