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Ivaylo L Dimitrov1, Feyzim V Hodzhaoglu, Dobryana P Koleva

  • 1Department of Phase Formation, Crystalline and Amorphous Materials, Rostislaw Kaischew Institute of Physical Chemistry, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., bl. 11, Sofia, 1113, Bulgaria, idimitrov@ipc.bas.bg.

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This study develops a novel method to measure lysozyme crystal nucleation rates by analyzing crystal presence in sampled solution volumes. The technique provides direct evaluation of nucleation parameters and is adaptable for various soluble substances.

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Area of Science:

  • Crystallization Science
  • Biophysical Chemistry
  • Materials Science

Background:

  • Crystal nucleation is a critical, stochastic process in crystallization.
  • Understanding nucleation kinetics is essential for controlling crystal formation.
  • Lysozyme crystallization is a model system for protein crystallization studies.

Purpose of the Study:

  • To investigate lysozyme crystal nucleation in quiescent solutions under constant supersaturation.
  • To develop and validate a novel experimental method for estimating crystal nucleation rates.
  • To determine key nucleation parameters including stationary nucleation rates and nucleus formation work.

Main Methods:

  • Utilized optical microscopy and discrete time sampling of small solution volumes.
  • Employed a probabilistic approach based on the stochastic nature of nucleation.
  • Fitted experimental data to a probability function for parameter evaluation.

Main Results:

  • Successfully estimated probabilities for crystal detection.
  • Directly evaluated stationary rates for lysozyme crystal nucleation.
  • Calculated nucleation parameters such as nucleus size and surface energy.

Conclusions:

  • The developed method offers a simple and adaptable approach for studying crystal nucleation.
  • The technique allows for direct evaluation of nucleation kinetics and related parameters.
  • This method can be applied to diverse soluble substances for nucleation studies.