Comparative three-dimensional imaging of living neurons with confocal and atomic force microscopy

McNally, Helen; Rajwa, Bartek; Sturgis, Jennifer; Robinson, J. Paul

doi:10.1016/j.jneumeth.2004.08.018

Cited by 35 publications

(31 citation statements)

References 24 publications

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“…The combination of CLSM with AFM has already been described in the literature [18][19][20]. For example, a comparison of CLSM and AFM images taken independently on similar (but not identical) areas of neuron samples has demonstrated the potential of combining the information obtained by both techniques [21]. One of the key innovations shown in the present study is to apply CLSM and AFM on exactly the same part of biofilm samples.…”

Section: Introductionmentioning

confidence: 84%

Towards chemical analysis of nanostructures in biofilms I: imaging of biological nanostructures

et al. 2008

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Due to their direct influence on the stability of bacterial biofilms, a better insight into the nanoscopic spatial arrangement of the different extracellular polymeric substances (EPS), e.g., polysaccharides and proteins, is important for the improvement of biocides and for process optimization in wastewater treatment and biofiltration. Here, the first application of a combination of confocal laser-scanning microscopy (CLSM) and atomic force microscopy (AFM) to the investigation of river-water biofilms and related biopolymers is presented. AFM images collected at selected areas of CLS micrographs dramatically demonstrate the heterogeneity of biofilms at the nanometer scale and the need for a chemical imaging method with nanoscale resolution. The nanostructures (e.g., pili, flagella, hydrocolloids, and EPS) found in the extracellular matrix are classified according to shape and size, which is typically 50-150 nm in width and 1-10 nm in thickness, and sets the demands regarding spatial resolution of a potential chemical imaging method. Additionally, thin layers of the polysaccharide alginate were investigated. We demonstrate that calcium alginate is a good model for the EPS architecture at the nanometer scale, because of its similar network-like structure.

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Section: Introductionmentioning

confidence: 84%

Towards chemical analysis of nanostructures in biofilms I: imaging of biological nanostructures

et al. 2008

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“…Indeed, recently the use of AFM was reported to emphasize various new applications in neurobiology (38).…”

Section: Discussionmentioning

confidence: 99%

Static magnetic fields affect cell size, shape, orientation, and membrane surface of human glioblastoma cells, as demonstrated by electron, optic, and atomic force microscopy

et al. 2006

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Background: It is common knowledge that static magnetic fields (SMF) do not interact with living cells; thus, fewer studies of SMF compared with variable magnetic fields are carried out. However, evidence demonstrated that SMF affect cellular structures. To investigate the effect of exposure to increasing doses of SMF on cell morphology, human glioblastoma cells were exposed to SMF ranging between 80 and 3,000 G (8 and 300 mT). Methods: Cell morphology of human glioblastoma cells, derived from a primary culture, was studied by electron and optic microscopy. FITC-phalloidin staining of actin filaments was also investigated. Finally, cell surface structure changes were detected by atomic force microscopy.Results: Scanning electron microscopy demonstrated a dose-dependent cell shape modification, progressive cell detachment, loss of the long villi, and appearance of membrane roughness and blebs. FITC-phalloidin staining confirmed the villi retention and cell dimension decrease. At

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“…Henderson et al, 1992;Rotsch et al, 1997;Schneider et al, 1997;Braet et al, 2001;McNally and Borgens, 2004;Pesen and Hoh, 2005). A small number of AFM studies investigated neurons, focusing on the cell body, axon, and synaptic vesicle structure (Parpura et al, 1993;Lal et al, 1995;Tojima et al, 2000;McNally and Borgens, 2004;Ricci et al, 2004;McNally et al, 2005); however, there was a limited attempt to study neuronal growth cones in detail.…”

Section: Introductionmentioning

confidence: 99%

High‐resolution analysis of neuronal growth cone morphology by comparative atomic force and optical microscopy

Grzywa

Lee

et al. 2006

J. Neurobiol.

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Neuronal growth cones are motile sensory structures at the tip of axons, transducing guidance information into directional movements towards target cells. The morphology and dynamics of neuronal growth cones have been well characterized with optical techniques; however, very little quantitative information is available on the three-dimensional structure and mechanical properties of distinct subregions. In the present study, we imaged the large Aplysia growth cones after chemical fixation with the atomic force microscope (AFM) and directly compared our data with images acquired by light microscopy methods. Constant force imaging in contact mode in combination with force-distant measurements revealed an average height of 200 nm for the peripheral (P) domain, 800 nm for the transition (T) zone, and 1200 nm for the central (C) domain, respectively. The AFM images show that the filopodial F-actin bundles are stiffer than surrounding F-actin networks. Enlarged filopodia tips are 60 nm higher than the corresponding shafts. Measurements of the mechanical properties of the specific growth cone regions with the AFM revealed that the T zone is stiffer than the P and the C domain. Direct comparison of AFM and optical data acquired by differential interference contrast and fluorescence microscopy revealed a good correlation between these imaging methods. However, the AFM provides height and volume information at higher resolution than fluorescence methods frequently used to estimate the volume of cellular compartments. These findings suggest that AFM measurements on live growth cones will provide a quantitative understanding of how proteins can move between different growth cone regions.

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Comparative three-dimensional imaging of living neurons with confocal and atomic force microscopy

Cited by 35 publications

References 24 publications

Towards chemical analysis of nanostructures in biofilms I: imaging of biological nanostructures

Towards chemical analysis of nanostructures in biofilms I: imaging of biological nanostructures

Static magnetic fields affect cell size, shape, orientation, and membrane surface of human glioblastoma cells, as demonstrated by electron, optic, and atomic force microscopy

High‐resolution analysis of neuronal growth cone morphology by comparative atomic force and optical microscopy

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