冷冻过程中洋葱细胞胞内冰晶的生长行为研究
|
摘要
细胞胞内冰的形成会导致严重的细胞损伤从而导致低温贮存中的诸多问题。生物组织保存前的预冷过程对胞内冰晶的形成具有重要影响。本文分别采用-1,-5,-10,-20,-50,-80℃/min的降温速率预冷洋葱表皮细胞至-7℃接种冰晶.采用高速摄像仪观察冰晶生长过程,发现了三种模式的胞内冰晶生长行为。模式A:细胞壁结晶后继续在边缘结晶;模式B:细胞壁结晶后,向中心扩散,横贯中心位置;模式C:中心首先结晶,扩散至细胞边缘。最后对三种不同过程的冰晶生长速率及细胞变形度进行了计算分析。
The formation of intracellular ice in the cell could lead to severe cell damage and many problems in cryopreservation. This paper investigates the effects of pre-cooling before ice seeding. The onion epidermal cells were pre-cooled at-1,-,5-10,-20,-50 and-80 ℃/min respectively and the ice is seeded as soon as the cryostage achieved-7 ℃. The high speed camera was used to observe the patterns in the location and formation of intracellular ice crystals and to evaluate the degree of cell deformation. The results showed that an increase in pre-cooling rate influenced the formation pattern of intracellular ice crystals. Mode A: ice crystal first appearing in the cytoderm and spreading to another cytoderm; Mode B: ice crystal spreading toward the center after cytoderm crystallization;Mode C: ice first appearing in the inner cell and spreading to cytoderm. Finally, the growth rate and cell deformability of ice crystals were calculated and analyzed in three different processes.
The formation of intracellular ice in the cell could lead to severe cell damage and many problems in cryopreservation. This paper investigates the effects of pre-cooling before ice seeding. The onion epidermal cells were pre-cooled at-1,-,5-10,-20,-50 and-80 ℃/min respectively and the ice is seeded as soon as the cryostage achieved-7 ℃. The high speed camera was used to observe the patterns in the location and formation of intracellular ice crystals and to evaluate the degree of cell deformation. The results showed that an increase in pre-cooling rate influenced the formation pattern of intracellular ice crystals. Mode A: ice crystal first appearing in the cytoderm and spreading to another cytoderm; Mode B: ice crystal spreading toward the center after cytoderm crystallization;Mode C: ice first appearing in the inner cell and spreading to cytoderm. Finally, the growth rate and cell deformability of ice crystals were calculated and analyzed in three different processes.
引文
[1]华泽钊.低温生物医学技术[M].北京:科学出版社,1994HUA Zezhao.Low-Temperature Biomedical Technology[M].Beijing:Science Press,1994
[2]Mansoori G A.Kinetics of Water Loss from Cells at Subzero Centigrade Temperatures[J].Cryobiology,1975,12(1):34-45
[3]Mazur P.Freezing of Living Cells:Mechanisms and Implications[J].American Journal of Physiology,1984,247(3Pt 1):125-142
[4]Leef J L,Mazur P.Physiological Response of Neurospora Conidia to Freezing in the Dehydrated,Hydrated,or Germinated State[J].Applied&Environmental Microbiology,1978,35(1):72-83
[5]Ninagawa T,Eguchi A,Kawamura Y,et al.A Study on Ice Crystal Formation Behavior at Intracellular Freezing of Plant Cells Using a High-Speed Camera[J].Cryobiology,2016,73(1):20-29
[6]杨戈尔,张爱丽,徐学敏,等.胞内冰晶形成(综述)[J].工程热物理学报,2007,28(z2):55-57YANG Geer,ZHANG Aili,XU Xuemin,et al.Formation of Intracellular Ice Crystals(Review)[J].Journal of Engineering Thermophysics,2007,28(z2):55-57
[7]Modlibowska I,Rogers W S.Freezing of Plant Tissues Under the Microscope[J].Journal of Experimental Botany,1955,6(18):384-391
[8]Asahina E.The Freezing Process of Plant Cell[J].Contributions from the Institute of Low Temperature Science,1956,10:83-126
[9]Stott S L,Karlsson J O M.Visualization of Intracellular Ice Formation Using High-Speed Video Cryomicroscopy[J].Cryobiology,2009,58(1):84-95
[10]张哲,赵静,田津津,等.冷冻-复温过程中葡萄细胞结晶变化研究[J].农业机械学报,2016,47(5):211-217ZHANG Zhe,ZHAO Jing,TIAN Jinjin,et al.Study on Crystallization Changes of Grape Cells During FreezingRestoration[J].Journal of Agricultural Machinery,2016,47(5):211-217
[11]Schneider C A,Rasband W S,Eliceiri K W.NIH Image to Image J:25 Years of Image Analysis[J].Nature Methods,2012,7(9):671-675
[12]高永毅.植物的细胞力学行为研究及在水果机械损伤研究中的应用[D].北京:中国农业大学,2003GAO Yongyi.Study on Plant Cytological Behavior and Its Application to Fruit Mechanical Damage[D].Beijng:China Agricultural University,2003
[2]Mansoori G A.Kinetics of Water Loss from Cells at Subzero Centigrade Temperatures[J].Cryobiology,1975,12(1):34-45
[3]Mazur P.Freezing of Living Cells:Mechanisms and Implications[J].American Journal of Physiology,1984,247(3Pt 1):125-142
[4]Leef J L,Mazur P.Physiological Response of Neurospora Conidia to Freezing in the Dehydrated,Hydrated,or Germinated State[J].Applied&Environmental Microbiology,1978,35(1):72-83
[5]Ninagawa T,Eguchi A,Kawamura Y,et al.A Study on Ice Crystal Formation Behavior at Intracellular Freezing of Plant Cells Using a High-Speed Camera[J].Cryobiology,2016,73(1):20-29
[6]杨戈尔,张爱丽,徐学敏,等.胞内冰晶形成(综述)[J].工程热物理学报,2007,28(z2):55-57YANG Geer,ZHANG Aili,XU Xuemin,et al.Formation of Intracellular Ice Crystals(Review)[J].Journal of Engineering Thermophysics,2007,28(z2):55-57
[7]Modlibowska I,Rogers W S.Freezing of Plant Tissues Under the Microscope[J].Journal of Experimental Botany,1955,6(18):384-391
[8]Asahina E.The Freezing Process of Plant Cell[J].Contributions from the Institute of Low Temperature Science,1956,10:83-126
[9]Stott S L,Karlsson J O M.Visualization of Intracellular Ice Formation Using High-Speed Video Cryomicroscopy[J].Cryobiology,2009,58(1):84-95
[10]张哲,赵静,田津津,等.冷冻-复温过程中葡萄细胞结晶变化研究[J].农业机械学报,2016,47(5):211-217ZHANG Zhe,ZHAO Jing,TIAN Jinjin,et al.Study on Crystallization Changes of Grape Cells During FreezingRestoration[J].Journal of Agricultural Machinery,2016,47(5):211-217
[11]Schneider C A,Rasband W S,Eliceiri K W.NIH Image to Image J:25 Years of Image Analysis[J].Nature Methods,2012,7(9):671-675
[12]高永毅.植物的细胞力学行为研究及在水果机械损伤研究中的应用[D].北京:中国农业大学,2003GAO Yongyi.Study on Plant Cytological Behavior and Its Application to Fruit Mechanical Damage[D].Beijng:China Agricultural University,2003