共查询到20条相似文献,搜索用时 31 毫秒
1.
P Perner A Rapp C Dressler L Wollweber J Beuthan K O Greulich M Hausmann 《Analytical cellular pathology》2002,24(2-3):89-100
The response of single breast cancer cells (cell line T-47D) to 17beta-estradiol (E(2)) under different concentrations was studied by using an instrument that allows to combine far-field light microscopy with high resolution scanning near-field (AFM/SNOM) microscopy on the same cell. Different concentrations of E(2) induce clearly different effects as well on cellular shape (in classical bright-field imaging) as on surface topography (atomic force imaging) and absorbance (near-field light transmission imaging). The differences range from a polygonal shape at zero via a roughly spherical shape at physiological up to a spindle-like shape at un-physiologically high concentrations. The surface topography of untreated control cells was found to be regular and smooth with small overall height modulations. At physiological E(2) concentrations the surfaces became increasingly jagged as detected by an increase in membrane height. After application of the un-physiological high E(2) concentration the cell surface structures appeared to be smoother again with an irregular fine structure. The general behaviour of dose dependent differences was also found in the near-field light transmission images. In order to quantify the treatment effects, line scans through the normalised topography images were drawn and a rate of co-localisation between high topography and high transmission areas was calculated. The cell biological aspects of these observations are, so far, not studied in detail but measurements on single cells offer new perspectives to be empirically used in diagnosis and therapy control of breast cancers. 相似文献
2.
Understanding the structural organization and distribution of proteins in biological cells is of fundamental importance in biomedical research. The use of conventional fluorescent microscopy for this purpose is limited due to its relatively low spatial resolution compared to the size of a single protein molecule. Atomic force microscopy (AFM), on the other hand, allows one to achieve single-protein resolution by scanning the cell surface using a specialized ligand-coated AFM tip. However, because this method relies on short-range interactions, it is limited to the detection of binding sites that are directly accessible to the AFM tip. We developed a method based on magnetic (long-range) interactions and applied it to investigate the structural organization and distribution of endothelin receptors on the surface of smooth muscle cells. Endothelin receptors were labeled with 50-nm superparamagnetic microbeads and then imaged with magnetic AFM. Considering its high spatial resolution and ability to “see” magnetically labeled proteins at a distance of up to 150 nm, this approach may become an important tool for investigating the dynamics of individual proteins both on the cell membrane and in the submembrane space. 相似文献
3.
N. N. van der Wel C. A. J. Putman S. J. T. van Noort B. G. de Grooth A. M. C. Emons 《Protoplasma》1996,194(1-2):29-39
Summary Atomic force microscopy (AFM) holds unique prospects for biological microscopy, such as nanometer resolution and the possibility of measuring samples in (physiological) solutions. This article reports the results of an examination of various types of plant material with the AFM. AFM images of the surface of pollen grains ofKalanchoe blossfeldiana andZea mays were compared with field emission scanning electron microscope (FESEM) images. AFM reached the same resolutions as FESEM but did not provide an overall view of the pollen grains. Using AFM in torsion mode, however, it was possible to reveal differences in friction forces of the surface of the pollen grains. Cellulose microfibrils in the cell wall of root hairs ofRaphanus sativus andZ. mays were imaged using AFM and transmission electron microscopy (TEM). Imaging was performed on specimens from which the wall matrix had been extracted. The cell wall texture of the root hairs was depicted clearly with AFM and was similar to the texture known from TEM. It was not possible to resolve substructures in a single microfibril. Because the scanning tip damaged the fragile cells, it was not possible to obtain images of living protoplasts ofZ. mays, but images of fixed and dried protoplasts are shown. We demonstrate that AFM of plant cells reaches resolutions as obtained with FESEM and TEM, but obstacles still have to be overcome before imaging of living protoplasts in physiological conditions can be realized.Abbreviations AFM
atomic force microscope
- FESEM
field emission scanning electron microscope
- PyMS
pyrolysis mass spectrometry
- TEM
transmission electron microscope 相似文献
4.
The application of atomic force microscopy to the study of living vertebrate cells in culture 总被引:8,自引:0,他引:8
Dvorak JA 《Methods (San Diego, Calif.)》2003,29(1):86-96
Atomic force microscopy (AFM), a relatively new variant of scanning probe microscopy developed for the material sciences, is becoming an increasingly important tool in other disciplines. In this review I describe in nontechnical terms some of the basic aspects of using AFM to study living vertebrate cells. Although AFM has some unusual attributes such as an ability to be used with living cells, AFM also has attributes that make its use in cell biology a real challenge. This review was written to encourage researchers in the biological and biomedical sciences to consider AFM as a potential (and potent) tool for their cell biological research. 相似文献
5.
Novel Combination of Atomic Force Microscopy and Epifluorescence Microscopy for Visualization of Leaching Bacteria on Pyrite 总被引:5,自引:0,他引:5
下载免费PDF全文
![点击此处可从《Applied microbiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Stefanie Mangold Kerstin Harneit Thore Rohwerder Günter Claus Wolfgang Sand 《Applied microbiology》2008,74(2):410-415
Bioleaching of metal sulfides is an interfacial process comprising the interactions of attached bacterial cells and bacterial extracellular polymeric substances with the surface of a mineral sulfide. Such processes and the associated biofilms can be investigated at high spatial resolution using atomic force microscopy (AFM). Therefore, we visualized biofilms of the meso-acidophilic leaching bacterium Acidithiobacillus ferrooxidans strain A2 on the metal sulfide pyrite with a newly developed combination of AFM with epifluorescence microscopy (EFM). This novel system allowed the imaging of the same sample location with both instruments. The pyrite sample, as fixed on a shuttle stage, was transferred between AFM and EFM devices. By staining the bacterial DNA with a specific fluorescence dye, bacterial cells were labeled and could easily be distinguished from other topographic features occurring in the AFM image. AFM scanning in liquid caused deformation and detachment of cells, but scanning in air had no effect on cell integrity. In summary, we successfully demonstrate that the new microscopic system was applicable for visualizing bioleaching samples. Moreover, the combination of AFM and EFM in general seems to be a powerful tool for investigations of biofilms on opaque materials and will help to advance our knowledge of biological interfacial processes. In principle, the shuttle stage can be transferred to additional instruments, and combinations of AFM and EFM with other surface-analyzing devices can be proposed. 相似文献
6.
Atomic force microscopy (AFM) has been used to study the micromechanical properties of biological systems. Its unique ability to function both as an imaging device and force sensor with nanometer resolution in both gaseous and liquid environments has meant that AFM has provided unique insights into the mechanical behaviour of tissues, cells and single molecules. As a surface scanning device, AFM can map properties such as adhesion and the Young's modulus of surfaces. As a force sensor and nanoindentor AFM can directly measure properties such as the Young's modulus of surfaces or the binding forces of cells. As a stress-strain gauge AFM can study the stretching of single molecules or fibres and as a nanomanipulator it can dissect biological particles such as viruses or DNA strands. The present paper reviews key research that has demonstrated the versatility of AFM and how it can be exploited to study the micromechanical behaviour of biological materials. 相似文献
7.
Atomic force and total internal reflection fluorescence microscopy for the study of force transmission in endothelial cells 总被引:9,自引:0,他引:9
下载免费PDF全文
![点击此处可从《Biophysical journal》网站下载免费的PDF全文](/ch/ext_images/free.gif)
This paper describes the combined use of atomic force microscopy (AFM) and total internal reflection fluorescence microscopy (TIRFM) to examine the transmission of force from the apical cell membrane to the basal cell membrane. A Bioscope AFM was mounted on an inverted microscope, the stage of which was configured for TIRFM imaging of fluorescently labeled human umbilical vein endothelial cells (HUVECs). Variable-angle TIRFM experiments were conducted to calibrate the coupling angle with the depth of penetration of the evanescent wave. A measure of cellular mechanical properties was obtained by collecting a set of force curves over the entire apical cell surface. A linear regression fit of the force-indentation curves to an elastic model yields an elastic modulus of 7.22 +/- 0. 46 kPa over the nucleus, 2.97 +/- 0.79 kPa over the cell body in proximity to the nucleus, and 1.27 +/- 0.36 kPa on the cell body near the edge. Stress transmission was investigated by imaging the response of the basal surface to localized force application over the apical surface. The focal contacts changed in position and contact area when forces of 0.3-0.5 nN were applied. There was a significant increase in focal contact area when the force was removed (p < 0.01) from the nucleus as compared to the contact area before force application. There was no significant change in focal contact coverage area before and after force application over the edge. The results suggest that cells transfer localized stress from the apical to the basal surface globally, resulting in rearrangement of contacts on the basal surface. 相似文献
8.
9.
Lewis W. Francis Paul D. Lewis Chris J. Wright R. Steve Conlan 《Biology of the cell / under the auspices of the European Cell Biology Organization》2010,102(2):133-143
AFM (atomic force microscopy) analysis, both of fixed cells, and live cells in physiological environments, is set to offer a step change in the research of cellular function. With the ability to map cell topography and morphology, provide structural details of surface proteins and their expression patterns and to detect pico‐Newton force interactions, AFM represents an exciting addition to the arsenal of the cell biologist. With the explosion of new applications, and the advent of combined instrumentation such as AFM—confocal systems, the biological application of AFM has come of age. The use of AFM in the area of biomedical research has been proposed for some time, and is one where a significant impact could be made. Fixed cell analysis provides qualitative and quantitative subcellular and surface data capable of revealing new biomarkers in medical pathologies. Image height and contrast, surface roughness, fractal, volume and force analysis provide a platform for the multiparameter analysis of cell and protein functions. Here, we review the current status of AFM in the field and discuss the important contribution AFM is poised to make in the understanding of biological systems. 相似文献
10.
Atomic force microscopy (AFM) is a specialised form of scanning probe microscopy, which was invented by Binnig and colleagues in 1986. Since then, AFM has been increasingly used to study biomedical problems. Because of its high resolution, AFM has been used to examine the topography or shape of surfaces, such as during the molecular imaging of proteins. This, combined with the ability to operate under known force regimes, makes AFM technology particularly useful for measuring intermolecular bond forces and assessing the mechanical properties of biological materials. Many of the constraints (e.g. complex instrumentation, slow acquisition speeds and poor vertical range) that previously limited the use of AFM in cell biology are now beginning to be resolved. Technological advances will enable AFM to challenge both confocal laser scanning microscopy and scanning electron microscopy as a method for carrying out three-dimensional imaging. Its use as both a precise micro-manipulator and a measurement tool will probably result in many novel and exciting applications in the future. In this article, we have reviewed some of the current biological applications of AFM, and illustrated these applications using studies of the cell biology of bone and integrin-mediated adhesion. 相似文献
11.
Ki Woo Kim 《Journal of microbiology (Seoul, Korea)》2016,54(11):703-708
Microorganisms, or microbes, can function as threatening pathogens that cause disease in humans, animals, and plants; however, they also act as litter decomposers in natural ecosystems. As the outermost barrier and interface with the environment, the microbial cell surface is crucial for cell-to-cell communication and is a potential target of chemotherapeutic agents. Surface ultrastructures of microbial cells have typically been observed using scanning electron microscopy (SEM) and atomic force microscopy (AFM). Owing to its characteristics of low-temperature specimen preparation and superb resolution (down to 1 nm), cryo-field emission SEM has revealed paired rodlets, referred to as hydrophobins, on the cell walls of bacteria and fungi. Recent technological advances in AFM have enabled high-speed live cell imaging in liquid at the nanoscale level, leading to clear visualization of cell-drug interactions. Platinum-carbon replicas from freeze-fractured fungal spores have been observed using transmission electron microscopy, revealing hydrophobins with varying dimensions. In addition, AFM has been used to resolve bacteriophages in their free state and during infection of bacterial cells. Various microscopy techniques with enhanced spatial resolution, imaging speed, and versatile specimen preparation are being used to document cellular structures and events, thus addressing unanswered biological questions. 相似文献
12.
13.
14.
苍耳柄锈菌三裂叶豚草专化型冬孢子萌发过程及萌发条件的研究 总被引:2,自引:1,他引:1
本文采用光学显微镜和扫描电子显微镜技术及荧光染色技术,对苍耳柄锈菌三裂叶豚草专化型Puccinia xanthii sp.ambrosiae-trifidae冬孢子的萌发过程和萌发条件进行了研究。结果表明:冬孢子堆成熟时突破寄主表皮外露;在寄主上冬孢子萌发时由上细胞顶部出现皱褶和帽状物,由帽状物下伸出担子。冬孢子的上细胞和下细胞都可萌发;冬孢子在水中于25℃2h即可萌发,24h后达到萌发高峰,萌发率为12%;温度20-25℃、相对湿度97%以上、pH5-7的条件利于冬孢子萌发,光照对冬孢子萌发没有影响,木糖和乳糖对冬孢子萌发有促进作用;无机氮源营养对冬孢子萌发有抑制作用。肌醇、烟酸、核黄素及三裂叶豚草叶汁对冬孢子萌发有促进作用。 相似文献
15.
Changes in the Endosperm Cell Walls of Two Datura Species before Radicle Protrusion 总被引:3,自引:2,他引:1
下载免费PDF全文
![点击此处可从《Plant physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
The possibility of an association between changes in cell walls of the micropylar portion of the endosperm and the induction of germination was explored in seeds of Datura ferox and Datura stramonium. The structure of the inner surface of the endosperm was studied by scanning electron microscopy and the composition of cell wall polysaccharides analyzed by gas chromatography and gas chromatography-mass spectrometry. Both scanning electron microscope images and chemical analysis showed changes in the micropylar portion of the endosperm in induced seeds before radicle protrusion. The inner surface of the endosperm appeared eroded, and in some areas, wall material seemed to be missing. The content of the main component of the cell wall polysaccharides, containing predominantly 4-linked mannose, decreased well before the emergence of the radicle through the endosperm. We propose that the degradation of a mannan type polysaccharide is an important factor in the reduction in mechanical strength of the endosperm, thus facilitating germination. 相似文献
16.
Arnaldo da Silva Junior Omar Teschke 《World journal of microbiology & biotechnology》2005,21(6-7):1103-1110
Summary Atomic force microscopy (AFM) images of living cells in physiological solution were used to monitor the different stages involved
in the interaction between Escherichia coli and the antimicrobial peptide PGLa. Damage on bacterial membranes was observed in the past using standard electron microscopy;
stiffness measurements and images scanned in physiological solution demonstrate the advantage of AFM for such studies. From
force versus separation curve measurements it is possible to determine the variation of the cellular stiffness. PGLa action
on components of the cell structure like the outer membrane, the bacterial pili, the peptidoglycan wall and the inner membrane
was determined by the comparison of AFM images of bacteria before and after PGLa addition. The interaction of Escherichia coli with PGLa in the culture medium has two stages. The first is characterized by the loss of surface stiffness and the formation
of micelles probably originating from the disruption of the outer membrane and the loss of the bacteria’s ability to adhere
to the substrates. In the second stage there is further damage, which resulted in total cell rupture. AFM images of bacteria
in air and surface roughness measurements were also used to estimate peptide damage. 相似文献
17.
M. S. Kuyukina I. O. Korshunova E. V. Rubtsova I. B. Ivshina 《Applied Biochemistry and Microbiology》2014,50(1):1-9
Atomic-force microscopy (AFM) is an efficient method for studying the surface ultrastructure and nanomechanical properties of biological objects, including microorganisms. A correctly selected method of microorganism immobilization that provides a strong attachment of cells on the surface of a biologically inert substrate and preservation of their native properties is important for AFM scanning in liquid media. Comparative characteristics of methods of microorganism immobilization applied in dynamic AFM studies are discussed in the review. Technologies of mechanical entrapment and chemical binding of cells to a substrate, as well as protein and immunospecific adsorption, are considered. 相似文献
18.
Yu. M. Efremov D. V. Bagrov E. V. Dubrovin K. V. Shaitan I. V. Yaminskii 《Biophysics》2011,56(2):257-267
We review the advances of the method of atomic force microscopy (AFM) for investigating the animal cells and analyze its development,
paying much attention to studies of living cells. We consider the specific features and tasks of AFM, and a number of special
AFM-based techniques. We discuss the choice of probe geometry for studies of animal cells, determination of cell adhesion
on substrate, mapping of the cell surface using chemically modified cantilevers, and analysis of the distribution of molecular
components inside the cell with the use of micro- and nanosurgical approaches, as well as combining AFM with optical and laser
scanning confocal microscopy, and the possible applications of AFM in biotechnology and medicine. 相似文献
19.
Establishment and characterization of human malignant lymphoma cell line derived from uterine cervix
Kiguchi K Ishiwata I Ishiwata C Ishiwata E Koshitaka Y Iwata M Okuda Y Kobayashi Y Wada Y Ishizuka B Ishikawa H 《Human cell》2005,18(1):53-58
Human uterine cervical malignant lymphoma (B-cell type) was cultured and the cell line (HIUML) was newly established. The HIUML cells were round in shape and had a tendency to make floating clusters. The cells had a smooth surface or protrusion on the margin of the cytoplasm, and proliferate in floatation. The population doubling time was about 32 hours and 42 or more passages were successfully observed in two years. The HIUML cells were not transplantable into nude mice but were successfully done in the cheek pouch of hamster with formation of malignant lymphoma. Epstein-Barr virus was detected in the HIUML cells. 相似文献
20.
Bacterial biofilms impair the operation of many industrial processes. Deinococcus geothermalis is efficient primary biofilm former in paper machine water, functioning as an adhesion platform for secondary biofilm bacteria. It produces thick biofilms on various abiotic surfaces, but the mechanism of attachment is not known. High-resolution field-emission scanning electron microscopy and atomic force microscopy (AFM) showed peritrichous adhesion threads mediating the attachment of D. geothermalis E50051 to stainless steel and glass surfaces and cell-to-cell attachment, irrespective of the growth medium. Extensive slime matrix was absent from the D. geothermalis E50051 biofilms. AFM of the attached cells revealed regions on the cell surface with different topography, viscoelasticity, and adhesiveness, possibly representing different surface layers that were patchily exposed. We used oscillating probe techniques to keep the tip-biofilm interactions as small as possible. In spite of this, AFM imaging of living D. geothermalis E50051 biofilms in water resulted in repositioning but not in detachment of the surface-attached cells. The irreversibly attached cells did not detach when pushed with a glass capillary but escaped the mechanical force by sliding along the surface. Air drying eliminated the flexibility of attachment, but it resumed after reimmersion in water. Biofilms were evaluated for their strength of attachment. D. geothermalis E50051 persisted 1 h of washing with 0.2% NaOH or 0.5% sodium dodecyl sulfate, in contrast to biofilms of Burkholderia cepacia F28L1 or the well-characterized biofilm former Staphylococcus epidermidis O-47. Deinococcus radiodurans strain DSM 20539(T) also formed tenacious biofilms. This paper shows that D. geothermalis has firm but laterally slippery attachment not reported before for a nonmotile species. 相似文献