Atomic force microscopy of BHK-21 cells: an investigation of cell fixation techniques

Moloney, Mary; McDonnell, L.; O’Shea, Helen

doi:10.1016/j.ultramic.2003.12.010

Cited by 51 publications

(44 citation statements)

References 22 publications

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“…ELISA detection of anti-fibroblast antibodies A cell-based ELISA was performed on fixed and unpermeabilised fibroblasts [12]. In order to adjust for interassay variability, the optical density value of a positive reference serum was arbitrarily defined as 100% of anti-fibroblast activity.…”

Section: Cell Culturementioning

confidence: 99%

Antibodies to fibroblasts in idiopathic and scleroderma-associated pulmonary hypertension

Tamby

Humbert²,

Guilpain³

et al. 2006

European Respiratory Journal

118

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The aim of the present study was to investigate the presence of anti-fibroblast antibodies in patients with idiopathic or scleroderma-associated pulmonary arterial hypertension (PAH) and healthy controls.PAH was documented by right-heart catheterisation (mean pulmonary artery pressure at rest .25 mmHg). Serum immunoglobulin (Ig)G and IgM reactivities of patients with idiopathic PAH (n535), scleroderma-associated PAH (n510), diffuse (n510) or limited cutaneous (n510) scleroderma without PAH and age-and sex-matched healthy individuals (n565) were analysed by cell-based ELISA and immunoblotting on normal human fibroblasts.As assessed by ELISA, 14 out of 35 (40%) patients with idiopathic PAH and three out of 10 (30%) patients with scleroderma-associated PAH expressed anti-fibroblast IgG antibodies. IgG from all individuals bound to one major 40-kDa protein band. IgG from patients with idiopathic PAH bound to two 25-and 60-kDa bands with a higher intensity than IgG from other individuals.In conclusion, immunoglobulin G anti-fibroblast antibodies are present in the serum of patients with pulmonary arterial hypertension. Immunoglobulin G from patients with idiopathic pulmonary arterial hypertension or scleroderma-associated pulmonary arterial hypertension express distinct reactivity profiles with fibroblasts antigens, suggesting distinct target antigens.

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Section: Cell Culturementioning

confidence: 99%

Antibodies to fibroblasts in idiopathic and scleroderma-associated pulmonary hypertension

Tamby

Humbert²,

Guilpain³

et al. 2006

European Respiratory Journal

118

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“…However, FEISEM ultrastructural analysis showed a definitely compromised cell organization where the cytoplasm and the nucleus were totally extracted. Moloney et al (2004) evaluated the efficiency of different fixatives on cell morphology by atomic force microscopy. They described ethanol/acetic acid-fixed cells as \volcano-like structures" corresponding to depressions inside cell bodies due to dehydration of the cells by the fixative.…”

Section: Discussionmentioning

confidence: 99%

Effect of Fixative on Chromatin Structure and DNA Detection

Falconi

Teti

Zago

et al. 2007

Microscopy Res & Technique

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In this study, we analyzed the chromatin ultrastructure in interphase cells after different chemical fixations. In light of the fact that there is little information regarding the fixation of biological samples in combination with molecular biology methods (such as DNA extraction and in situ hybridization methods) we analyzed the ultrastructure of chromatin in interphase cells fixed with different fixatives and tested under the same conditions for both DNA extraction and in situ hybridization. The results showed that, among the different combinations and concentrations we analyzed, the solution of 4% paraformaldehyde/0.1% glutaraldehyde was the best compromise in order to achieve a well-preserved morphology, successful DNA extraction, and specific signaling of in situ hybridization, suggesting a low interference of this fixative with the chromatin organization.

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“…In general, the fixation procedures, including chemical or physical methods, would change cell morphologies and mechanical properties. Aldehyde-based fixatives are useful for conserving cell or bacterial morphologies [60,61]. Nonetheless, it is very possible that different samples require different notes: This figure is adopted from Ref.…”

Section: Cell Membrane Responses To Chemical Stimulationsmentioning

confidence: 99%

Mechanoprofiling on membranes of living cells with atomic force microscopy and optical nano-profilometry

Lee

Wang

Lee

2017

Advances in Physics: X

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Membrane topography of living cells has been considered as an effective parameter that reflects cellular statuses. With the improvements in spatial and temporal resolutions of various measurement techniques, the changes of membrane topography in response to various external stimulations in the culture environments can be accurately recorded. Membrane roughness is a useful parameter to evaluate the changes in membrane topography. At present, atomic force microscopy (AFM) is the most common technique to measure the membrane topography and roughness of living cells. On the other hand, with the non-contact profiling capability, many optical techniques are also employed in this field of research. This review briefly introduces the evolution and applications of AFM on measuring membrane roughness. Meanwhile, quantifying the cell membrane topography and roughness with a non-scanning, non-contact, and label-free optical technique, non-interferometric wide-field optical profilometry (NIWOP), is also presented.

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Atomic force microscopy of BHK-21 cells: an investigation of cell fixation techniques

Cited by 51 publications

References 22 publications

Antibodies to fibroblasts in idiopathic and scleroderma-associated pulmonary hypertension

Antibodies to fibroblasts in idiopathic and scleroderma-associated pulmonary hypertension

Effect of Fixative on Chromatin Structure and DNA Detection

Mechanoprofiling on membranes of living cells with atomic force microscopy and optical nano-profilometry

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