Scale-like cantilever cell traps

Liu, Boyin; Fu, Jianzhong; Somers, Anthony; Muradoglu, Murat; Ng, Tuck Wah

doi:10.1039/c3ra46123h

Cited by 1 publication

(2 citation statements)

References 32 publications

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“…In a previous study, adhesion force was applied to quantity antibiotic and peptidoglycan interaction [14], and combining the same tip functionalization protocols with the current study has great potential for intracellular drug localization studies. Together with recently developed approaches for improved bacterial cell collection within specific regions [26,76] and correlative imaging [77][78][79], we expect that the complete FIB-AFM platform will help in gaining deeper insights of bacteria-drug interactions to develop potential strategies for combating multi-drug resistance.…”

Section: Discussionmentioning

confidence: 99%

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In situprobing the interior of single bacterial cells at nanometer scale

Liu

Uddin

et al. 2014

Nanotechnology

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We report a novel approach to probe the interior of single bacterial cells at nanometre resolution by combining focused ion beam (FIB) and atomic force microscopy (AFM). After removing layers of pre-defined thickness in the order of 100 nm on the target bacterial cells with FIB milling, AFM of different modes can be employed to probe the cellular interior under both ambient and aqueous environments. Our initial investigations focused on the surface topology induced by FIB milling and the hydration effects on AFM measurements, followed by assessment of the sample protocols. With fine-tuning of the process parameters, in situ AFM probing beneath the bacterial cell wall was achieved for the first time. We further demonstrate the proposed method by performing a spatial mapping of intracellular elasticity and chemistry of the multi-drug resistant strain Klebsiella pneumoniae cells prior to and after it was exposed to the 'last-line' antibiotic polymyxin B. Our results revealed increased stiffness occurring in both surface and interior regions of the treated cells, suggesting loss of integrity of the outer membrane from polymyxin treatments. In addition, the hydrophobicity measurement using a functionalized AFM tip was able to highlight the evident hydrophobic portion of the cell such as the regions containing cell membrane. We expect that the proposed FIB-AFM platform will help in gaining deeper insights of bacteria-drug interactions to develop potential strategies for combating multi-drug resistance.

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Section: Discussionmentioning

confidence: 99%

“…After treatment with 1 mL of 2.5% glutaraldehyde in phosphate buffered saline for 20 min, the samples were washed two times and re-suspended in Milli-Q water. A recent approach for improved cell collection at specific regions was applied in this process [26].…”

Section: Cell Growth and Harvestingmentioning

confidence: 99%