Applications of gray-level variation detection method to intracellular ice formation

Area of Science:

• Cell biology
• Cryobiology
• Biophysics

Background:

• Intracellular ice formation (IIF) is a primary cause of cell death during freezing.
• IIF impedes light penetration, darkening images and complicating analysis.
• Cell deformation often accompanies IIF, necessitating a combined analytical approach.

Purpose of the Study:

• To develop a unified method for analyzing intracellular ice formation (IIF) and accompanying cell deformation.
• To quantify the relationship between gray-level variations and cell displacement during freezing.

Main Methods:

• A normalized parameter C was defined, correlating gray-level variations with pixel displacement.
• The method analyzes changes in gray-level for each pixel within the region of interest.
• The process of IIF was studied in onion epidermal cells and 293T cells.

Main Results:

• The normalized parameter C effectively reflects gray-level changes associated with IIF.
• The method accounts for both IIF-induced image darkening and cell deformation.
• Analysis demonstrated the applicability of the method to different cell types.

Conclusions:

• The developed method provides a comprehensive approach to studying intracellular ice formation.
• This technique enhances the understanding of cell freezing injury by integrating optical and displacement data.
• The findings are relevant for cryopreservation and understanding cellular responses to freezing.