Abstract
Preparation of isolated cells and microorganisms for ultrastructural examination always provides a challenge in terms of adequate immobilization of the cells and prevention of subsequent sample loss and damage during various steps of sample processing. Using a positively charged nylon membrane substrate we demonstrate that it is possible to easily immobilize and retain a sample of isolated cells in culture for a wide variety of microscopy-based techniques. Radiolabelled E. coli cells when immobilized on the charged membrane were seen to be highly resistant to detachment when subjected to the normal sample processing procedures associated with microscopy. In contrast cells on regular millipore membranes were rapidly lost during sample preparation. We demonstrate the utility of charged nylon membranes for a wide variety of microscopy based analysis including scanning and transmission electron microscopy (SEM and TEM), atomic force microscopy (AFM), TEM based immunogold labelling, laser confocal microscopy and SEM based elemental analysis.
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Acknowledgements
The skilful technical assistance of Mr. Anil Theophilus is gratefully acknowledged. R. Dhiman was the recipient of CSIR research fellowship. This manuscript is IMTECH communication number No. 05/2006.
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Raje, M., Dhiman, R., Majumdar, S. et al. Charged nylon membrane substrate for convenient and versatile high resolution microscopic analysis of Escherichia coli & mammalian cells in suspension culture. Cytotechnology 51, 111–117 (2006). https://doi.org/10.1007/s10616-006-9027-2
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DOI: https://doi.org/10.1007/s10616-006-9027-2