Abstract
We report a three dimensional (3D) quantitative visualization of a mammalian mitochondrion by coherent x-ray diffractive imaging (CXDI) using synchrotron radiation. The internal structures of a mitochondrion from a mouse embryonic fibroblast cell line (NIH3T3) were visualized by tomographic imaging at approximately 60 nm resolution without the need for sectioning or staining. The overall structure consisted of a high electron density region, composed of the outer and inner membranes and the cristae cluster, which enclosed the lower density mitochondrial matrix. The average mass density of the mitochondrion was about 1.36 g/cm3. Sectioned images of the cristae reveal that they have neither a baffle nor septa shape but were instead irregular. In addition, a high resolution, about 14 nm, 2D projection image was captured of a similar mitochondrion with the aid of strongly scattering Au reference objects. Obtaining 3D images at this improved resolution will allow CXDI to be an effective and nondestructive method for investigating the innate structure of mitochondria and other important life supporting organelles.
Highlights
Over the last few decades, there has been enormous effort to visualize the internal structures of cellular organelles utilizing a variety of microscopy techniques such as optical, fluorescence, and electron microscopies[1,2,3,4,5,6,7,8,9,10]
We demonstrate the feasibility of Coherent x-ray diffractive imaging (CXDI) for imaging the internal structure of a mammalian mitochondrion three dimensionally
We illustrate the feasibility of visualizing unstained and unsectioned mitochondria by applying CXDI to a mitochondrion of NIH3T3 which is a standard mammalian fibroblast cell line
Summary
Over the last few decades, there has been enormous effort to visualize the internal structures of cellular organelles utilizing a variety of microscopy techniques such as optical, fluorescence, and electron microscopies[1,2,3,4,5,6,7,8,9,10]. The resolution of conventional optical microscopies is fundamentally limited to several hundred nanometers due to long wavelengths used, following Abbe’s principle This is not sufficient to observe most sub-cellular structures. Much higher spatial resolution is available in electron microscopies but thick specimens have to be sectioned which disrupts their internal structure, and in addition most of them need to be stained with heavy metals, which complicates image interpretation[11,12,13]. All these methods suffer from the aberration issues caused by various types of lenses involved in microscopes. A high resolution 2D projection image was obtained using strongly scattering Au reference objects[32, 33] that amplify the diffraction signal
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