N.m.r. imaging of intact biological systems

Internal images of structured objects may be obtained with n.m.r. by labelling component parts with different magnetic field strengths and therefore recognizably different n.m.r. frequencies. A linear field gradient generates a one-dimensional projection of nuclear density and a variety of techniques are employed to manipulate this one-dimensional probe to yield internal images in two and three dimensions. In the past few years, n.m.r. imaging, sometimes also called zeugmatography or spin mapping, has been applied progressively to provide proton images of small phantoms, fruit, vegetables and small animals, and finally to in vivo imaging of the human body; it promises to provide a valuable means of interior investigation of intact biological systems generally. For medical imaging the method is non-invasive, does not use ionizing radiations, appears to be without hazard and penetrates bony cavities without attenuation. Furthermore, other n.m.r. parameters, for example, relaxation times and fluid flow, may also be mapped; there is evidence that the relaxation times from tumours are significantly longer than those from corresponding normal tissue. Effort to date has mostly been concentrated on proton n.m.r., but some work has been done with other nuclei. Three examples are shown of n.m.r. images of intact biological systems: a fruit, an animal and a human system. The discussion includes the quantitative nature of the images, tissue discrimination, the relation between the resolution in the image and image acquisition time, attenuation and phase shift of the r.f. field in the biological tissue, and magnets suitable for n.m.r. imaging. In principle, all conventional n.m.r. techniques can be combined with n.m.r. methods in order to investigate heterogeneous systems. Overhauser imaging is briefly discussed.     

The Wellcome Foundation lecture, 1981. Nuclear magnetic resonance imaging in medicine: physical principles

In recent years nuclear magnetic resonance (n.m.r.) has become a means of providing excellent images of the interior of the human body which are proving useful in medical practice. The development of n.m.r. imaging, much of which was pioneered in Britain, is outlined. Proton image resolution of human anatomy is comparable with X-ray computed tomography images, but without the hazard of ionizing radiation. There is improved soft tissue discrimination and pathological contrast through the basic imaging parameters of the proton density and the relaxation times T1 and T2, whose differences from one tissue to another are exploited by use of appropriate radiofrequency pulse sequences. Images may be obtained directly of transverse, coronal and sagittal sections of the head and body. Single slices or multiple slices may be imaged and imaging may be done in three dimensions. The lecture describes the more important imaging techniques and gives illustrative examples of images obtained. The efficient use of time in n.m.r. imaging is discussed, particularly mentioning the multiecho-multislice procedure and the development of real-time n.m.r. imaging. Magnetic field strengths in current use for proton n.m.r. imaging range from 0.02 to 2 T. At the lower end of the range resistive magnets are used, while for higher fields superconducting magnets are needed. A considerable improvement in image quality is obtained by use of special receiver coils.     

The Wellcome Foundation lecture, 1984. Nuclear magnetic resonance imaging in medicine: medical and biological applications and problems

From early biological work and the first T1 nuclear magnetic resonance (n.m.r.) animal image in 1974, whole-body patient images, by using a two-dimensional Fourier transform method were achieved in Aberdeen in 1980 with a 0.04 T vertical resistive magnet. Different pulse sequences produce images dependent by different amounts on proton density, T1 and T2, and for clinical work it is advantageous to use more than one pulse sequence to image pathology. The slow improvement of spatial resolution with increasing standing magnetic field strength is discussed and information on the T1 and T2 contrast dependence is reviewed: it suggests that the gains from high fields may be less than believed hitherto. Electrocardiogram gating can be used to produce moving images of the beating heart; blood flow can be imaged and surface radiofrequency coils are used for improved detail. N.m.r. imaging has considerable potential for studying response to therapy; mental states and dementia; tissue generation; discriminating body fat and body fluids. Other nuclei such as 23Na can be imaged and the potential to image fluorine-labelled pharmaceuticals could be very exciting; n.m.r. contrast agents are now being developed. Images formed from T1 values measured for each pixel are very useful for diagnosis, but the numerical values themselves are less valuable for distinctive pathological identification. With 15 companies manufacturing n.m.r. imagers and over 200 in use in hospitals, the technique is rapidly becoming established in diagnostic clinical practice and some typical uses are presented.     

N.m.r. spectroscopy: A non-invasive tool for studying intracellular processes

The development of nuclear magnetic resonance (n.m.r.) spectroscopy as a non-invasive, non-destructive tool for the study of metabolic processes in cells and tissues is reviewed. Although n.m.r. measurements are subject to some limitations, e.g. low sensitivity and the need for special probes for in vivo work, the quality of n.m.r. instruments is steadily improving and wide bore spectrometers have been introduced. The article reviews in vivo n.m.r. studies of suspended and immobilized cells and also gives examples of the use of solid state n.m.r. Plant cell suspensions and tissues are good systems for study by n.m.r. as the P_i resonances from different compartments in the cell can be easily identified, and are important indicators of cellular metabolism. The literature on the use of magnetization transfer and on techniques for the study of enzyme catalysed reactions is also reviewed. Future applications of n.m.r. in examining biotechnological processes are discussed.     

Fluorescent probes in membrane studies.

A number of spectroscopic techniques are suitable for studying biological membranes. Of these, fluorescence has the sensitivity and time resolution for following membrane events associated with nerve excitation. In this paper, the nature of the information derived from measurements of the fluorescence properties of externally introduced chromophores in membranes is examined. In particular, the locations of various probes are described on the basis of nuclear magnetic resonance (n.m.r.) experiments in model situations. Then the motional characteristics of the probe molecules (rotation and diffusion) are discussed. Finally experiments designed to relate the detailed observations that can be made in lipid bilayers using n.m.r. and fuorescence measurements to those (more limited in nature) that can be made in membranes are described.     

Field focusing n.m.r. (FONAR) and the formation of chemical images in man

The first proposals for n.m.r. scanning in medical diagnosis was made by Damadian (1971a; 1972) and were followed by Lauterbur (1973). Damadian's method of scanning used the principle that the forced precessions of a nuclear magnetization under radio frequency (r.f.) driving field specify the conditions for obtaining spatial resolution of the signal producing domains of a nuclear resonance sample. Sufficient coupling of the nuclear spins to the radiation field to produce a signal detectable by r.f. spectroscopy requires that the stringent Bohr frequency condition, hv = microH0/I, be met. It became possible to construct, with the aid of direct current auxiliary coils, a small volume, called the resonance aperture, inside the applied static field of the magnetic resonance experiment. The correct value of H0 for the applied frequency is restricted to this aperture. The technique (Damadian 1972) was developed to provide a method for non-surgically detecting chemical abnormalities in the diseased organs of patients (Damadian 1971a). The first n.m.r. scans of normal patients and of those with malignant disease are discussed.     

N.m.r. studies of metabolism in perfused organs

Several metabolites and intracellular pH in intact organs can be studied in a non-destructive manner by phorphorus nuclear magnetic resonance (31P n.m.r.). This possibility was demonstrated by us nearly five years ago. Since then we have developed the appropriate physiological techniques and improved the n.m.r. method for the study of animal hearts and kidneys. Here we described measurements aimed at clarifying three problesm. (1) Having measured the enzyme-catalysed fluxes between phosphocreatine and ATP by the method of saturation transfer n.m.r., we examine the relations between energy supply and heart rate in the isolated perfused rat heart. (2) We describe experiments to establish the validity of the perfusion model. For the first time, we report 31P n.m.r. measurements of an in vivo rat heart and compare the results with those obtained for the perfused rat heart. (3) Ischaemia and metabolism in rabbit kidneys is investigated to establish the relation between functional and metabolic recovery after a renal transplant operation.     

Some biophysical applications of motional contrast in n.m.r. microscopy

The principal advantage of the n.m.r. imaging method lies in the specific contrasts which are available. In this work we describe the use of velocity and diffusion contrast methods in biophysical applications and at microscopic spatial resolution. In the first example, involving water-protein interactions, the relationship between water self-diffusion and water concentration, as measured using pulsed gradient spin echo n.m.r., is shown. It is demonstrated that this relationship can be used to provide a water concentration image. The result is compared with the conventional proton density and transverse relaxation maps. The next example concerns the use of dynamic n.m.r. microscopy to obtain water diffusion and velocity maps for wheat grain in vivo. Finally we suggest how the method may be used in the study of polymer-water interactions in an unusual adjunct to conventional polymer self-diffusion studies.     

医学领域的磁共振成像革命——2003年诺贝尔生理及医学奖主要工作介绍

在磁场中 ,自旋的原子核会吸收频率与其自旋频率相同的电磁波 ,使自身能量增加 ,发生能级跃迁 ,当原子核迁移回原能级时 ,就会把多余的能量以电磁波的形式释放出来 ,称为核磁共振 (NMR) .磁共振成像(MRI)利用这一原理 ,依据所释放的能量在物质内部不同结构环境中不同的衰减 ,通过外加梯度磁场检测所发射出的电磁波 ,即可得知构成这一物体原子核的位置和种类 ,据此可以绘制成物体内部的结构图像 .将这种技术用于人体内部结构的成像 ,就产生出一种革命性的医学诊断工具 .快速变化的梯度磁场的应用 ,大大加快了磁共振成像的速度 ,使该技术在临床诊断、科学研究的应用成为现实 ,极大地推动了医学、神经生理学和认知神经科学的迅速发展     

In vivo biomicroscopy of the skin with high-resolution magnetic resonance imaging and high frequency ultrasound.

Noninvasive imaging and characterization of the skin is of great interest in dermatology. In order to get relevant diagnostic information, high-resolution imaging techniques have to be applied. Ultrasonic imaging is a potential method for this purpose where the special requirements concerning the spatial resolution make it essential to apply high frequency ultrasound (HFUS). Alternatively, magnetic resonance imaging (MRI), being a very promising imaging modality, also shows the perspective of becoming a valuable diagnostic tool in dermatology. However, to account for the small dimensions of the structures under observation, very specialized system designs have to be developed. In this paper, a HFUS imaging system working in the 50 MHz and 100 MHz range is applied for high-resolution skin imaging. Furthermore, a commercial MRI-system was equipped with specially designed low noise rf (radio frequency) coils with minimized volume, and customized imaging sequences were applied to optimize the signal-to-noise ratio. With HFUS and high-resolution magnetic resonance (HR-MR) imaging complementary imaging techniques for in vivo biomicroscopy of the skin are available.     

光学透明技术在植物多尺度成像中的应用

光学透明技术是一种通过各种化学试剂,将原本不透明的生物样本实现透明化,并在光学显微镜下深度成像的技术。结合多种光学显微成像新技术,光学透明技术可对整个组织进行成像和三维重建,深度剖析生物体内部空间特征与形成机制。近年来,多种植物光学透明技术和多尺度成像技术被陆续研发,并取得了丰硕的研究成果。该文综述了生物体光学透明技术的基本原理和一些新技术,重点介绍基于光学透明技术开发的新型成像方法及其在植物成像与细胞生物学中的应用,为后续植物整体、组织或器官的透明、成像与三维重构及功能研究提供理论依据和技术支持。     

Experimental considerations in implementing a whole body multiple sensitive point nuclear magnetic resonance imaging system

The multiple sensitive point technique on n.m.r. imaging is discussed in terms of the mode of n.m.r. excitation and detection, provision of field gradients and the methods of data processing. The design criteria of an instrument to implement this and related imaging methods at whole body size are described and particular attention is paid to techniques employed to increase the versatility and ease of use of the apparatus.     

Artificial intelligence in image reconstruction: The change is here

Innovations in CT have been impressive among imaging and medical technologies in both the hardware and software domain. The range and speed of CT scanning improved from the introduction of multidetector-row CT scanners with wide-array detectors and faster gantry rotation speeds. To tackle concerns over rising radiation doses from its increasing use and to improve image quality, CT reconstruction techniques evolved from filtered back projection to commercial release of iterative reconstruction techniques, and recently, of deep learning (DL)-based image reconstruction. These newer reconstruction techniques enable improved or retained image quality versus filtered back projection at lower radiation doses. DL can aid in image reconstruction with training data without total reliance on the physical model of the imaging process, unique artifacts of PCD-CT due to charge sharing, K-escape, fluorescence x-ray emission, and pulse pileups can be handled in the data-driven fashion. With sufficiently reconstructed images, a well-designed network can be trained to upgrade image quality over a practical/clinical threshold or define new/killer applications. Besides, the much smaller detector pixel for PCD-CT can lead to huge computational costs with traditional model-based iterative reconstruction methods whereas deep networks can be much faster with training and validation. In this review, we present techniques, applications, uses, and limitations of deep learning-based image reconstruction methods in CT.     

Automatic system for 3D reconstruction of the chick eye based on digital photographs

The geometry of anatomical specimens is very complex and accurate 3D reconstruction is important for morphological studies, finite element analysis (FEA) and rapid prototyping. Although magnetic resonance imaging, computed tomography and laser scanners can be used for reconstructing biological structures, the cost of the equipment is fairly high and specialised technicians are required to operate the equipment, making such approaches limiting in terms of accessibility. In this paper, a novel automatic system for 3D surface reconstruction of the chick eye from digital photographs of a serially sectioned specimen is presented as a potential cost-effective and practical alternative. The system is designed to allow for automatic detection of the external surface of the chick eye. Automatic alignment of the photographs is performed using a combination of coloured markers and an algorithm based on complex phase order likelihood that is robust to noise and illumination variations. Automatic segmentation of the external boundaries of the eye from the aligned photographs is performed using a novel level-set segmentation approach based on a complex phase order energy functional. The extracted boundaries are sampled to construct a 3D point cloud, and a combination of Delaunay triangulation and subdivision surfaces is employed to construct the final triangular mesh. Experimental results using digital photographs of the chick eye show that the proposed system is capable of producing accurate 3D reconstructions of the external surface of the eye. The 3D model geometry is similar to a real chick eye and could be used for morphological studies and FEA.     

Superresolution microscopy for microbiology

This review provides a practical introduction to superresolution microscopy from the perspective of microbiological research. Because of the small sizes of bacterial cells, superresolution methods are particularly powerful and suitable for revealing details of cellular structures that are not resolvable under conventional fluorescence light microscopy. Here we describe the methodological concepts behind three major categories of superresolution light microscopy: photoactivated localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM) and stimulated emission‐depletion (STED) microscopy. We then present recent applications of each of these techniques to microbial systems, which have revealed novel conformations of cellular structures and described new properties of in vivo protein function and interactions. Finally, we discuss the unique issues related to implementing each of these superresolution techniques with bacterial specimens and suggest avenues for future development. The goal of this review is to provide the necessary technical background for interested microbiologists to choose the appropriate superresolution method for their biological systems, and to introduce the practical considerations required for designing and analysing superresolution imaging experiments.     

Ultrasonographic contrast-agent imaging of sub-millimeter vessel structures with spatial compounding: in vitro analyses.

In clinical diagnostics, ultrasonographic contrast-agent imaging gives access to medical parameters such as perfusion and vascularization. In addition to the artifacts that are typical for ultrasonic imaging, e.g., speckle noise and depth-dependent sensitivity and resolution, contrast-agent imaging shows more pronounced depth dependence and may suffer from shadowing artifacts that arise from high attenuation of the ultrasound waves by the contrast agent at high concentrations. By imaging an object from different viewing angles in one 2D image plane and summing the images obtained (spatial compounding), image quality can be increased and artifacts can be suppressed. In the present study, we combined both techniques to overcome the limitations of contrast-agent imaging. We used a commercially available ultrasound scanner and a custom-made high-precision mechanical system to rotate the ultrasound transducer fully around the object under investigation. Using this set-up, ultrasound data were acquired in reflection mode to generate a 360 degrees compound scan of a flow-mimicking phantom supplied with contrast agent.     

Studies of the biochemistry of contracting and relaxing muscle by the use of 31P n.m.r. in conjunction with other techniques

When n.m.r. is applied to suitable chosen biological problems it yields a wealth of fundamental information unmatched by any other technique. By means of 31P n.m.r. we have studied intact living muscle at rest, during contraction and during recovery from contraction. Phosphocreatine, ATP, inorganic phosphate, phosphorylated intermediaries of glycolysis, pH and the binding of Mg2+ to ATP are observed directly in the spectra. From the spectra can be calculated the concentration of free ADP, the free energy change of ATP hydrolysis, the production of lactic acid and the total ATP turnover. Changes in these quantities can thus be followed continuously in vivo and we have shown how they are related to the decline in force development and to the slowing of relaxation that occur during fatigue. Similar methods have been applied to study the control of glycolysis.     

Selective spin decoupling in the J-resolved two-dimensional 1H n.m.r. spectra of proteins.

Methods were developed where selective homonuclear spin decoupling is used for the identification of the spin systems of individual amino acid residues in J-resolved two-dimensional high field ¹H n.m.r. spectra of proteins. Experiments with the basic pancreatic trypsin inhibitor are shown to illustrate the practical application of these new techniques.     

N.m.r. studies of red cells

Recent n.m.r. studies of intact red cells are described. With 1H n.m.r. the normal high resolution spectra of red cells, even at high fields, are relatively uninformative because the very large number of resonances from the cells merge into a broad envelope. If a simple 90-tau-180 degree spin echo pulse sequence is used, however, many resonances can all be resolved. These include signals from haemoglobin histidines, glutathione, lactate and pyruvate. 13C and 31P signals have also been seen with a spectrometer converted to observe these nuclei essentially simultaneously. N.m.r. is well suited to monitor the time course of events after a perturbation of the cell system. Lactate increase, glutathione recovery after oxidation and alkylation of glutathione by iodoacetate can all be observed directly in red cell suspensions by means of 1H spin echo n.m.r. This method has also been used to measure isotope exchange (1H-2H) of lactate and of pyruvate at both the C-3 and the C-2 positions, and some of these exchange rates can be interpreted in terms of the activity of specific enzymes in the cells. 1H spin echo n.m.r. has also been used to obtain information about the transport rates of small molecules into cells. By means of the 13C/31P spectrometer and [13C-1] glucose, the 13C enrichment of 2,3-diphosphoglycerate (2,3-DPG) can be monitored at the same time as the levels of 2,3-DPG, ATP and inorganic phosphate are observed by 31P n.m.r.     

High-performance electron tomography of complex biological specimens

We have evaluated reconstruction methods using smooth basis functions in the electron tomography of complex biological specimens. In particular, we have investigated series expansion methods, with special emphasis on parallel computation. Among the methods investigated, the component averaging techniques have proven to be most efficient and have generally shown fast convergence rates. The use of smooth basis functions provides the reconstruction algorithms with an implicit regularization mechanism, very appropriate for noisy conditions. Furthermore, we have applied high-performance computing (HPC) techniques to address the computational requirements demanded by the reconstruction of large volumes. One of the standard techniques in parallel computing, domain decomposition, has yielded an effective computational algorithm which hides the latencies due to interprocessor communication. We present comparisons with weighted back-projection (WBP), one of the standard reconstruction methods in the areas of computational demand and reconstruction quality under noisy conditions. These techniques yield better results, according to objective measures of quality, than the weighted backprojection techniques after a very few iterations. As a consequence, the combination of efficient iterative algorithms and HPC techniques has proven to be well suited to the reconstruction of large biological specimens in electron tomography, yielding solutions in reasonable computation times.