Structural and elemental characterization of heart cells grown in a collagen matrix*1

LeFurgey, Ann; Hawkey, L.A.; Ingram, Peter; Lieberman, Melvyn

doi:10.1016/1047-8477(91)90061-z

Cited by 10 publications

(2 citation statements)

References 22 publications

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“…In these studies a variety of experimental systems are being utilized to address questions fundamental to cell physiology and pathophysiology; these systems include protozoa (Leishmania major) , cultured embryonic chick heart cells (LeFurgey et al, , 1991a, a suspension of proximal kidney tubules (LeFurgey et al, 1986b(LeFurgey et al, , 1991bSpencer et al, 1990aSpencer et al, , 1991, isolated single skeletal muscle fibres (Nassar et al, 1986;Ingram et al, 1989), whole heart preparations (Sommer et al, 1988b), and nasal epithelia . Our approach to investigating the roles of Ca, Na, K and other elements in cell function is, of necessity, multidisciplinary, for cell function involves complex interactions of metabolism, membrane transport, electrical properties, structure and element compartmentation.…”

Section: Experimental Rationale and Protocolsmentioning

confidence: 99%

Real‐time quantitative elemental analysis and mapping: microchemical imaging in cell physiology

LeFurgey

Davilla

Kopf

et al. 1992

Journal of Microscopy

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K E Y w ORDS. X-ray microanalysis, microchemical microscopy, biological cryosections, electron microscopy, X-ray mapping, cell physiology, computer imaging. S U M M A R YRecent advances in widely available microcomputers have made the acquisition and processing of digital quantitative X-ray maps of one to several cells readily feasible. Here we describe a system which uses a graphics-based microcomputer to acquire spectrally filtered X-ray elemental image maps that are fitted to standards, to display the image in real time, and to correct the post-acquisition image map with regard to specimen drift. Both high-resolution quantitative energy-dispersive X-ray images of freeze-dried cyrosections and low-dose quantitative bright-field images of frozenhydrated sections can be acquired to obtain element and water content from the same intracellular regions. T h e software programs developed, together with the associated hardware, also allow static probe acquisition of data from selected cell regions with spectral processing and quantification performed on-line in real time. In addition, the unified design of the software program provides for off-line processing and analysing by several investigators at microcomputers remote from the microscope. The overall experimental strategy employs computer-aided imaging, combined with static probes, as an essential interactive tool of investigation for biological analysis. This type of microchemical microscopy facilitates studies in cell physiology and pathophysiology which focus on mechanisms of ionic (elemental) compartmentation, i.e. structurefunction correlation at cellular and subcellular levels; it allows investigation of intracellular concentration gradients, of the heterogeneity of cell responses to stimuli, of certain fast physiological events in uiuo at ultrastructural resolution, and of events occurring with low incidence or involving cell-to-cell interactions.

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Section: Experimental Rationale and Protocolsmentioning

confidence: 99%

Real‐time quantitative elemental analysis and mapping: microchemical imaging in cell physiology

LeFurgey

Davilla

Kopf

et al. 1992

Journal of Microscopy

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“…ESI and EELS imaging create new challenges for optimal biological specimen preparation for the analysis of diffusible elements (COllieX, 1986;Simon and Heng, 1988). Potential problems of movement of ionic forms of non-protein bound elements may be eliminated by cryofixation and freeze drying (Godleski et al, 1990;LeFurgey et al, 1989).…”

Section: Discussionmentioning

confidence: 99%

Contribution of osmium tetroxide to the image quality and detectability of iron in cells studied by electron spectroscopic imaging and electron energy loss spectroscopy

Stearns

Katler

Godleski

1994

Microscopy Res & Technique

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The detection of elemental distributions within ultrastructural cellular components presents a number of challenges. There are many technical questions that need to be resolved including optimal fixation protocols. Another is the impact of heavy metals, such as osmium tetroxide (OsO4), on the detectability of other elements when OsO4 is used in chemical fixation protocols for biological samples. OsO4 was examined by varying its concentrations from 0% to 1% and time of fixation from 5 to 30 minutes with hamster alveolar macrophages. The morphological quality of cellular images observed and the detectability of iron using electron spectroscopic imaging (ESI) and electron energy loss spectroscopy (EELS) were evaluated. One percent OsO4 for 30 minutes in the chemical fixation protocol enhances the quality of the ESI and does not interfere with the ESI or EELS signal of iron. Positive results from both methods indicate the presence of the specific element. The loss of 59Fe during the chemical fixation procedure was also studied. Less than 10% was lost during the primary fixation step, but minimal losses occurred through dehydration, embedding, and sectioning. Careful technical assessment of the presence of an element as well as factors which might interfere with its detection is an important step in the application of any analytical microscopic technique.

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Collagen gels populated with rat neonatal heart cells can be used for optical recording of rhythmic contractions which also show ECG-like potentials

1994

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Neonatal heart cells populated collagen gels show rhythmic contractions which can be recorded optically and electrically. Optical recordings revealed two populations of rhythmically contracting gels: 1) highly coherent contracting collagen gels with normally distributed contraction interval times and contraction amplitudes, and 2) irregularly contracting gels with a multi-modal distribution of contraction interval times and amplitudes. The irregularly contracting gels were shown to be 'semi regular', which means that a short contraction interval was preferentially followed by another short interval. The volume of the collagen gel during the contraction decreased, and our calculations indicate that the myocytes expel 3-10 times their own volume from the gel. Changes in electrical potential were observed depending on the location of the electrodes. These electrical, ECG-like changes in potential were maximal when one electrode was placed in the centre and the other at the edge of the gel. The results of this study indicate that myocyte-populated collagen gels are a very promising system for studies of electrophysiology and coherent contractions.

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Structural and elemental characterization of heart cells grown in a collagen matrix*1

Cited by 10 publications

References 22 publications

Real‐time quantitative elemental analysis and mapping: microchemical imaging in cell physiology

Real‐time quantitative elemental analysis and mapping: microchemical imaging in cell physiology

Contribution of osmium tetroxide to the image quality and detectability of iron in cells studied by electron spectroscopic imaging and electron energy loss spectroscopy

Collagen gels populated with rat neonatal heart cells can be used for optical recording of rhythmic contractions which also show ECG-like potentials

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