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

Paper Details

Research Paper 01/10/2016
Views (878)
current_issue_feature_image
publication_file

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.
Copyright Statement: Copyright 2016; The Author(s).
License: CC BY-NC 4.0

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.

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

Related Articles

Muscle type and meat quality of local chickens according to preslaughter transport conditions and sex in Benin

Assouan Gabriel Bonou*, Finagnon Josée Bernice Houéssionon, Kocou Aimé Edenakpo, Serge Gbênagnon Ahounou, Chakirath Folakè Arikè Salifou, Issaka Abdou Karim Youssao, Int. J. Biosci. 27(6), 241-250, December 2025.

Effects of micronutrients and timing of application on the agronomic and yield characteristics of cucumber (Cucumis sativus)

Princess Anne C. Lagcao, Marissa C. Hitalia*, Int. J. Biosci. 27(6), 214-240, December 2025.

Response of different soybean varieties to phosphorus fertilizer microdosing and rhizobium inoculation in the sub-humid zone of Northern Benin

Pierre G. Tovihoudji*, Kamarou-Dine Seydou, Lionel Zadji, Sissou Zakari, Valerien A. Zinsou, Int. J. Biosci. 27(6), 201-213, December 2025.

On-farm validation of black soldier fly larvae meal as a sustainable replacement for shrimp meal in rainbow trout diets in the mid hills of Nepal

Ishori Singh Mahato, Krishna Paudel*, Sunita Chand, Anshuka Bhattarai, Int. J. Biosci. 27(6), 189-200, December 2025.

Insect fauna associated with Cucumis sativus (Cucurbitales: Cucurbitaceae) in Parakou, A cotton-growing area of central Benin

Lionel Zadji*, Mohamed Yaya, Roland Bocco, Prudencia M. Tovignahoua, Abdou-Abou-Bakari Lassissi, Raphael Okounou Toko, Hugues Baimey, Leonard Afouda, Int. J. Biosci. 27(6), 175-188, December 2025.

First record of two hymenopteran species, Brachymeria excarinata Gahan (Chalcididae) and Pteromalus sp. (Pteromalidae), as hyperparasitoids of Diadegma insulare in Senegal

Babacar Labou*, Etienne Tendeng, Mamadou Diatte, El hadji Sérigne Sylla, Karamoko Diarra, Int. J. Biosci. 27(6), 167-174, December 2025.

Hepatoprotective and antinociceptive effects of terpinolene in streptozotocin-induced diabetic peripheral neuropathic rats

Ravishankar Sarumathi, Muthukumaran Preethi, Chandrasekaran Sankaranarayanan*, Int. J. Biosci. 27(6), 156-166, December 2025.