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Cryo-electron Microscopy01:28

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Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
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Preparation of Binary and Ternary Deep Eutectic Systems
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Natural deep eutectic systems, an emerging class of cryoprotectant agents.

Ana Rita Jesus1, Liane Meneses1, Ana Rita C Duarte1

  • 1LAQV-REQUIMTE, Campus da Caparica, Monte da Caparica, 2825-149, Caparica, Portugal.

Cryobiology
|May 11, 2021
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Summary

Natural deep eutectic systems (NADES) show promise as cryoprotectant agents (CPAs) for cell preservation. These biocompatible compounds offer effective cryoprotection with low toxicity, presenting a viable alternative to conventional agents.

Keywords:
BiomaterialsCryobiologyCryopreservationFreeze-thawingNatural deep eutectic systemsNatural metabolites

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Area of Science:

  • Cryobiology
  • Biochemistry
  • Cell Biology

Background:

  • Cryoprotectant agents (CPAs) are crucial for preserving cells during freezing.
  • Conventional CPAs like DMSO can exhibit toxicity.
  • There is a need for safer and effective alternatives in cryopreservation.

Purpose of the Study:

  • To evaluate natural deep eutectic systems (NADES) as potential CPAs.
  • To assess the cytotoxicity and efficacy of NADES in cell cryopreservation.
  • To explore NADES as alternatives to traditional cryoprotectants.

Main Methods:

  • Preparation of NADES from natural primary metabolites found in cold-adapted animals.
  • Cytotoxicity assessment of NADES on L929 cells.
  • Evaluation of NADES as CPAs for L929 and HacaT cell lines after freeze-thawing.
  • Comparison of NADES performance against Me₂SO.

Main Results:

  • NADES demonstrated low cytotoxicity towards L929 cells at high concentrations (1-2 M).
  • L929 cells showed high tolerance to 10% (w/v) NADES compared to Me₂SO.
  • NADES exhibited similar cryoprotective effects to Me₂SO for L929 cells.
  • Significant improvement in post-thawing recovery was observed for HacaT cells using NADES.
  • NADES do not require removal after thawing and can function as CPAs at -20 °C.

Conclusions:

  • NADES are effective and safe cryoprotectant agents.
  • These systems offer advantages such as ease of use and effectiveness at lower temperatures.
  • NADES represent a promising alternative for cryobiology applications.