Integration of confocal and atomic force microscopy images

Kondra, Shripad; Laishram, Jummi; Ban, Jelena; Migliorini, Elisa; Foggia, Valentina Di; Lazzarino, Marco; Torre, Vincent; Ruaro, Maria Elisabetta

doi:10.1016/j.jneumeth.2008.09.034

Cited by 33 publications

(26 citation statements)

References 28 publications

(33 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For a systematic measurement of single cell stiffness, we improved the method that has been used previously to study the structural details of subcellular compartments, such as growth cones, by combining AFM with immunocytochemistry (Kondra et al, 2009). We dissociated cortical cells from each of the different stages of development and plated the cells on a grid-lined glass-bottomed dish in order to measure stiffness using AFM (Fig.…”

Section: Resultsmentioning

confidence: 99%

Systematic profiling of spatiotemporal tissue and cellular stiffness in the developing brain

et al. 2014

View full text Add to dashboard Cite

Accumulating evidence implicates the significance of the physical properties of the niche in influencing the behavior, growth and differentiation of stem cells. Among the physical properties, extracellular stiffness has been shown to have direct effects on fate determination in several cell types in vitro. However, little evidence exists concerning whether shifts in stiffness occur in vivo during tissue development. To address this question, we present a systematic strategy to evaluate the shift in stiffness in a developing tissue using the mouse embryonic cerebral cortex as an experimental model. We combined atomic force microscopy measurements of tissue and cellular stiffness with immunostaining of specific markers of neural differentiation to correlate the value of stiffness with the characteristic features of tissues and cells in the developing brain. We found that the stiffness of the ventricular and subventricular zones increases gradually during development. Furthermore, a peak in tissue stiffness appeared in the intermediate zone at E16.5. The stiffness of the cortical plate showed an initial increase but decreased at E18.5, although the cellular stiffness of neurons monotonically increased in association with the maturation of the microtubule cytoskeleton. These results indicate that tissue stiffness cannot be solely determined by the stiffness of the cells that constitute the tissue. Taken together, our method profiles the stiffness of living tissue and cells with defined characteristics and can therefore be utilized to further understand the role of stiffness as a physical factor that determines cell fate during the formation of the cerebral cortex and other tissues.

show abstract

Section: Resultsmentioning

confidence: 99%

Systematic profiling of spatiotemporal tissue and cellular stiffness in the developing brain

et al. 2014

View full text Add to dashboard Cite

show abstract

“…To apply these methods to the images derived from different microscopes we have implemented our Registration program using Matlab7.1 (The MathWorks Inc. http://www.mathworks.com) a standard tool common in the computer vision and biomedical community for statistical analysis and production of figures and images of analyzed data and results. For a more detailed mathematical description of the different transformations and description of the program see Kondra et al, 2009, Appendix 1, 2 and 3). Corresponding points or contours in AFM image and confocal image were marked by hand using a friendly graphical user interface developed for this particular purpose (Figure 3).…”

Section: Characterization Of Embryonic Stem (Es) Neuronal Differentiamentioning

confidence: 99%

“…This problem can be solved by marking many points along the contour of the structure in both images. The contours are then interpolated so that both of them contain the same number of equally spaced points (for mathematical description of transformation see Kondra et al, 2009). An example of this method is illustrated in Figure 5.…”

Section: Characterization Of Embryonic Stem (Es)mentioning

confidence: 99%

Characterization of Embryonic Stem (ES) Neuronal Differentiation Combining Atomic Force, Confocal and DIC Microscopy Imaging

Ruaro¹,

Ban²,

Torre³

2011

Embryonic Stem Cells - Differentiation and Pluripotent Alternatives

Self Cite

View full text Add to dashboard Cite

“…Requiring no special sample preparation, AFM was quickly adapted for studying cells including living and fixed ones: it has been used to reveal structural information in a wide variety of cells and tissues [16][17][18]. In the cancer research field, AFM was used fundamentally for the exploration of cancer-cell morphology [19] and cell elasticity [20] as AFM could easily measure strain and local elastic properties of living cells during physiological processes and in response to anticytoskeletal drugs [21][22][23][24][25].…”

Section: Introductionmentioning

confidence: 99%

Multimicroscopic study of curcumin effect on fixed nonmalignant and cancerous mammalian epithelial cells

Saab¹,

Estephan²,

Bec³

et al. 2011

Journal of Biophotonics

View full text Add to dashboard Cite

The morphology changes, in particular the organization of microtubules in mammalian nonmalignant HMEC 184A1 and cancerous MCF-7 cells during curcumin treatment have been investigated by utilizing multiphoton, fluorescence, and atomic force (AFM) microscopies. Fluorescence microscopy reveals formation of ring-like structures of microtubules circumscribing the nuclear area in HMEC 184A1 cells after treatment, while in MCF-7 cells, no important changes were observed. Topography analyses of fixed HMEC 184A1 and MCF-7 before and after treatment with curcumin were performed using AFM and the effect of the employed cells' fixation method was investigated on MCF-7 cells. Due to its indepth optical sectioning capacity multiphoton microscopy provided valuable complementary information on curcumin's effect on both cells' types. Combining information provided by AFM and optical fluorescence and biphoton microscopes allows us to gain a better understanding of the cells and their curcumin-induced changes, especially for microtubules which are the main target of antitumor chemotherapy treatments. Our multimicroscopic study demonstrates that 6 h incubation with curcumin does not induce significant modifications in the interphase microtubules in the malignant MCF7cell, whereas it has measurable effects on those of the nonmalignant HMEC 184A1 cells, revealing also morphology modifications over the nuclear area of these cells.

show abstract

Integration of confocal and atomic force microscopy images

Cited by 33 publications

References 28 publications

Systematic profiling of spatiotemporal tissue and cellular stiffness in the developing brain

Systematic profiling of spatiotemporal tissue and cellular stiffness in the developing brain

Characterization of Embryonic Stem (ES) Neuronal Differentiation Combining Atomic Force, Confocal and DIC Microscopy Imaging

Multimicroscopic study of curcumin effect on fixed nonmalignant and cancerous mammalian epithelial cells

Contact Info

Product

Resources

About