Evaluation of changes in erythrocytes using atomic force microscopy and scanning electron microscopy (death from hypothermia)

In medicine, the issues of hypothermia and “revival” of the body remain open. Insufficient study of the features of the mechanisms of dying at ultra-low temperatures gives grounds to supplement the understanding of the cellular and molecular mechanisms of the process of fatal hypothermia. The aim of this study was to evaluate the morphology of erythrocytes in fatal hypothermia at extremely low temperatures (below -40°C) using atomic force microscopy and scanning electron microscopy. The object of the study was blood smears of persons who died of hypothermia and of gunshot wounds. The study was conducted using the atomic force microscope NT-MDT, SolverNext model, in the semi-contact mode and the high-resolution scanning electron microscope JSM-7800F. In the man who died of hypothermia in conditions of ultra-low temperatures (-42°C), whose smear was made within 1 day, red blood cells were characterized by minimal changes than in the dead from a gunshot wound. The discocytic form of red blood cells with a flat surface was preserved, but we found an increase in the depth of the central concavity of red blood cells, which may be associated with an increase in the volume of red blood cells. It was found that even a small difference in body temperature (∆ 2.°C) at the time of blood collection affects the ratio of different forms of red blood cells after complete thawing: the lower the temperature, the more pronounced degenerative forms of red blood cells. After complete thawing of the corpse for 4 days, all red blood cells (100 %) had a different degree of dysmorphy, a flattened surface, an increase in the diameter and heterogeneity and plasma roughness, indicating signs of hemolysis.

erythrocytes, death from hypothermia, atomic force microscopy, scanning electron microscopy

1. Palmiere, C., Mangin, P. Postmortem biochemical investigations in hypothermia fatalities / C. Palmiere, P. Mangin // International Journal of Legal Medicine. - 2013. - № 2/127. - P. 267-276.

2. Алексеев, Р.З. Изменение общих клинических параметров и показателей сердечно сосудистой системы при холодовой травме у собак / Р.З. Алексеева // Якутский медицинский журнал. - 2017. - № 1 (57). - С. 54-56.

3. Palmiere C., Biochemical markers of fatal hypothermia / C. Palmiere, Bardy D. Bardy, Letovanec I. Letovanec, et al. // Forensic Science International. - 2013. - № 10 (226). - Т (1-3). - Р. 54-61. DOI: 10.1016/j.forsciint.2012.12.007. Epub 2013 Jan 10.

4. Мороз, В.В. Строение и функция эритроцита в норме и при критических состояниях / В.В. Мороз, А.М. Голубев, А.В. Афанасьев и др. // Общая реаниматология. - 2012. - Т.8. - № 1. - С. 52-60.

5. Кидалов, В.Н. К вопросу о физиологической значимости изменений формы, ультраструктуры и флуоресценции эритроцитов периферической крови, трансформирующихся в эхиноциты / В.Н. Кидалов, Н.И. Сясин, А.А. Хадарцев // Вестник новых медицинских технологий. - 2005. - № 2 (12). - С. 6-9.

6. Berling, C. The RBC morphological dependence of the RBC disaggregability / C. Berling, Lacombe C. Lacombe, Lelie`vre J. C. Lelie`vre et al. // Biorheology. - 1988. - № 5. - Р. 791-798.

7. Parthasarathi, K., Lipowsky, N.N. Capillary recruitment in response to tissue hypoxia and its dependence on red blood cell deformability // K. Parthasarathi, N.N. Lipowsky // American Journal Physiology. - 1999. - № 6 (277). - P. 2145-2157.

8. Бархина, Т.Г. Патология мембран форменных элементов крови при заболеваниях в экспериментах / Т.Г. Бархина, Г.М. Никитина, М.М. Бархина и др. // Успехи современного естествознания. - 2006. - № 6. - С. 64-66.

9. Вычисление объема эритроцитов при анализе данных атомно-силовой микроскоп [Электронный ресурс] // Фундаментальные исследования. - 2013. - № 1. - С. 181-184. Режим доступа: https://e.mail.ru/attachment/15552924170000001012/0;1?xemail=lucy_morgan97 %40mail.ru

10. Ross, P.D., Minton, A.P. Hard quasispherical model for the viscosity of hemoglobin solutions / P.D. Ross, A.P. Minton // Biochemical and Biophysycal Research Communications. - 1977. - № 4 (76). - P. З971 - 3976.