Evaluation of cells integrity using different fixation time by scanning electron microscopy

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Research Paper 01/10/2016
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Evaluation of cells integrity using different fixation time by scanning electron microscopy

M. Y. Ida Muryany, R. Nor Fadilah, M. Y. Ina Salwany, A. R. Ghazali, M. Zamri Saad, M. A. Kaswandi, A. Z. Sahalan, H. L. Hing
Int. J. Biosci.9( 4), 236-241, October 2016.
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Abstract

The study aims to evaluate the integrity and structural changes of two intestinal cells adhered with Lactobacillus sp. in different fixation time. HT-29 and CCD-18Co intestinal cells with Lactobacillus sp. were fixed with 2.5% glutaraldehyde in 0.1 M phosphate buffer for two, four, six and 12 hours at room temperature. Our study revealed that the different fixation time affects and change the integrity, viability and durability of cells. HT-29 cells structure remain intact even after 12 hours fixation while CCD-18Co cells remain intact at two and four hour fixation time whereas at six hour cells also remain intact except cilia structure not clearly seen on surface of the cell. However, at 12 hours fixation, CCD-18Co cells were completely broken and degraded. Lactobacillus sp. cells remains stable in both cell lines, showing that the bacteria cells were unaffected with various fixation time. The present study suggested fixation time is important as an aware different cell has different endurance and structural integrity.

VIEWS 42

Allen TD. 1983. The application of scanning electron microscopy to cells in culture: selected methodologies. Scanning electron microscopy 4, 1963–1972.

An Y, Friedman R. 2000. Handbook of Bacterial Adhesion. Principles, Methods and Applications. Humana Press Inc. Totowa, 1-27 p.

Bozzola JJ, Russel LD. 1999. Electron microscopy: Principles and techniques for biologists. Sudbury: Jones and Barlett Publishing Ltd., 48–71 p.

Chang DE, Smalley DJ,Tucker DL, Leatham MP, Norris WE, Stevenson SJ, Anderson AB, Grissom JE, Laux DC, Cohen SP, Conway T. 2004. Proceedings of the National Academy of Sciences of the United States of America 101, 7427.

Eddie W, Filip B, Hans D, Celien V, Ger K, Steven WM, Olde D, Maartje AJ van den B, Bart DG, Cees HCD, Chise T, Peter F. 2010. Fixation methods for electron microscopy of human and other liver. World Journal of Gastroenterology 16(23), 2851-2866. http://dx.doi.org/10.3748/wjg/16.23.2851-2866

Eltoum I, Fredenburgh J, Myers RB, Grizzle WE. 2001. Introduction to the theory and practice of fixation of tissues. Journal of Histotechnology 24, 173 -190. 

Gomide Jr. MH, Elton VS, Marcos M, Isabel CB. 2004. Use of scanning electron microscopy for the evaluation of intestinal epithelium integrity. Revista brasileira de zootecnia 33, 1500-1505. http://dx.doi.org/10.1590/S/33.1500-1505

Hayat MA. 2000. Principles and techniques of electron microscopy: Biological applications. 4th ed. Cambridge University Press, 400–431 p.

Mahta M, Michelle CA. 2010. An in vitro study on bacterial growth interactions and intestinal epithelial cell adhesion characteristics of probiotic combinations. Current Microbiology 60, 327-335. http://dx.doi.org/10.1007/s00284-009-9545-1

Neděla V, Tihlaříková E, Hampl A, Sedláčková M. 2012. SEM and ESEM observation of stem cells: Comparative study of human embryonic stem cell morphology. Imaging and microscopy 4, 32–34.

Samaranayake LP. 1996. Essential microbiology for dentistry. Churchill Livingstone, New York, p. 45-58.

Wurzinger-mayer A, Shipway JR, Kristof A, Scwaha T, Cragg SM, Wanninger A. 2014. Developmental dynamics of myogenesis in the shipworm Lyro pedicellatus (Mollusca: Bivalva). Frontiers in Zoology 11, 90. http://dx.doi.org/10.1186/s12983-014-0090-9