Abstract
Reconstruction of neural circuits at the microscopic scale of individual neurons and synapses, also known as connectomics, is an important challenge for neuroscience. While an important motivation of connectomics is providing anatomical ground truth for neural circuit models, the ability to decipher neural wiring maps at the individual cell level is also important in studies of many neurodegenerative diseases. Reconstruction of a neural circuit at the individual neuron level requires the use of electron microscopy images due to their extremely high resolution. Computational challenges include pixel-by-pixel annotation of these images into classes such as cell membrane, mitochondria and synaptic vesicles and the segmentation of individual neurons. State-of-the-art image analysis solutions are still far from the accuracy and robustness of human vision and biologists are still limited to studying small neural circuits using mostly manual analysis. In this chapter, we describe our image analysis pipeline that makes use of novel supervised machine learning techniques to tackle this problem.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Suzuki K, Horiba I, Sugie N (2003) Neural edge enhancer for supervised edge enhancement from noisy images. IEEE Trans Pattern Anal Mach Intell 25:1582–1596
Suzuki K, Horiba I, Sugie N, Nanki M (2004) Extraction of left ventricular contours from left ventriculograms by means of a neural edge detector. IEEE Trans Med Imag 23:330–339
Sporns O, Tononi G, Ktter R (2005) The human connectome: A structural description of the human brain. PLoS Comput Biol 1:e42
Briggman KL, Denk W (2006) Towards neural circuit reconstruction with volume electron microscopy techniques. Curr Opin Neurobiol 16:562–570
Mishchenko Y (2008) Automation of 3D reconstruction of neural tissue from large volume of conventional serial section transmission electron micrographs. J Neurosci Methods 176(2):276–289
Anderson J, Jones B, Yang J-H, Shaw M, Watt C, Koshevoy P, Spaltenstein J, Jurrus E, U V K, Whitaker R, Mastronarde D, Tasdizen T, Marc R (2009) A computational framework for ultrastructural mapping of neural circuitry. PLoS Biol 7(3):e74
Mishchenko Y, Hu T, Spacek J, Mendenhall J, Harris KM, Chklovskii DB (2010) Ultrastructural analysis of hippocampal neuropil from the connectomics perspective. Neuron 67:1009–1020
Cardona A, Saalfeld S, Preibisch S, Schmid B, Cheng A, Pulokas J, Tomančák P, Hartenstein V (2010) An integrated micro- and macroarchitectural analysis of the Drosophila brain by computer-assisted serial section electron microscopy. PLoS Biol 8(10):e1000502
Bock DD, Lee W-C, Kerlin AM, Andermann ML, Hood G, Wetzel AW, Yurgenson S, Soucy ER, Kim HS, Reid RC (2011) Network anatomy and in vivo physiology of visual cortial neurons. Nature 471:177–182
Briggman KL, Helmstaedter M, Denk W (2011) Wiring specificity in the direction-selectivity circuit of the retina. Nature 471:183–188
Marc RE, Jones BW, Watt CB, Vazquez-Chona F, Vaughan DK, Organisciak DT (2008) Extreme retinal remodeling triggered by light damage: implications for age related macular degeneration. Mol Vis 14:782–806
Marc RE, Jones BW, Watt CB, Strettoi E (2003) Neural remodeling in retinal degeneration. Progr Retin Eye Res 22:607–655
Sutula T (2002) Seizure-induced axonal sprouting: assessing connections between injury, local circuits, and epileptogenesis. Epilepsy Current 2:86–91
Koyama R, Yamada MK, Fujisawa S, Katoh-Semba R, Matsuki N, Ikegaya Y (2004) Brain-derived neurotrophic factor induces hyperexcitable reentrant circuits in the dentate gyrus. J Neurosci 24:7215–7224
Xiao YP, Wang Y, Felleman DJ (2003) A spatially organized representation of colour in macaque cortical area v2. Nature 421(6922):535–539
Minsky M (1961) Microscopy apparatus. U.S. Patent number 301,467, December 1961
Denk W, Strickler JH, Webb WW (1990) Two-photon laser scanning microscopy. Science 248:73–76
Egner A, Hell SW (2005) Fluorescence microscopy with super-resolved optical sections. Trends Cell Biol 15:207–215
Rust MJ, Bates M, Zhuang X (2006) Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (storm). Nat Meth 3:793–796
Betzig E, Patterson G, Sougrat R, Lindwasser O, Olenych S, Bonifacino J, Davidson M, Lippincott-Schwartz J, Hess H (2006) Imaging intracellular fluorescent proteins at nanometer resolution. Science 313(5793):1642–1645
White JG, Southgate E, Thomson JN, Brenner S (1986) The structure of the nervous system of the nematode caenorhabditis elegans. Philos Trans R Soc Lond B Biol Sci 314(1165):1–340
Hall DH, Russell RL (1991) The posterior nervous system of the nematode caenorhaditis elegans: Serial reconstruction of identified neurons and complete pattern of synaptic interactions. J Neurosci 11(1):1–22
Chen BL, Hall DH, Chklovskii DB (2006) Wiring optimization can relate neuronal structure and function. Proc Natl Acad Sci USA 103(12):4723–4728
Chklovskii DB, Vitaladevuni S, Scheffer LK (2010) Semi-automated reconstruction of neural circuits using electron microscopy. Curr Opin Neurobiol 20(5):667–675
Anderson JR, Jones BW, Watt CB, Shaw MV, Yang JH, Demill D, Lauritzen JS, Lin Y, Rapp KD, Mastronarde D, Koshevoy P, Grimm B, Tasdizen T, Whitaker R, Marc RE (2011) Exploring the retinal connectome. Mol Vis 17:355–379
Varshney LR, Chen BL, Paniagua E, Hall DH, Chklovskii DB (2011) Structural properties of the Caenorhabditis elegans neuronal network. PLoS Comput Biol 7(2):e1001066
Deerinck TJ, Bushong EA, Thor A, Ellisman MH (2010) NCMIR methods for 3D EM: A new protocol for preparation of biological specimens for serial block face scanning electron microscopy. Microscopy and Microanalysis Meeting, Portland, OR, 1–5 August 2010
Hayworth K, Kasthuri N, Schalek R, Lichtman J (2006) Automating the collection of ultrathin serial sections for large volume TEM reconstructions. Microsc Microanal 12(2):86–87
Tasdizen T, Koshevoy P, Grimm BC, Anderson JR, Jones BW, Watt CB, Whitaker RT, Marc RE (2010) Automatic mosaicking and volume assembly for high-throughput serial-section transmission electron microscopy. J Neurosci Meth 193(1):132–144
Saalfeld S, Cardona A, Hartenstein V, Tomančák P (2010) As-rigid-as-possible mosaicking and serial section registration of large ssTEM datasets. Bioinformatics 26(12):i57–i63
Tasdizen T, Koshevoy P, Grimm B, Anderson J, Jones B, Watt C, Whitaker R, Marc R (2010) Automatic mosaicking and volume assembly for high-throughput serial-section transmission electron microscopy. J Neurosci Meth 193:132–144
Anderson J, Mohammed S, Grimm B, Jones B, Koshevoy P, Tasdizen T, Whitaker R, Marc R (2011) The viking viewer for connectomics: scalable multi-user annotation and summarization of large volume data sets. J Microscopy 241:13–28
Knott G, Marchman H, Wall D, Lich B (2008) Serial section scanning electron microscopy of adult brain tissue using focused ion beam milling. J Neurosci 28(12):2959–2964
Soto GE, Young SJ, Martone ME, Deerinck TJ, Lamont S, Carragher BO, Hama K, Ellisman MH (1994) Serial section electron tomography: A method for three-dimensional reconstruction of large structures. NeuroImage 1(3):230–243
Chen X, Winters CA, Reese TS (2008) Life inside a thin section: Tomography. J Neurosci 28(38):9321–9327
Hama K, Arii T, Katayama E, Marton M, Ellisman MH (2004) Tri-dimensional morphometric analysis of astrocytic processes with high voltage electron microscopy of thick golgi preparations. J Neurocytol 33:277–285. doi:10.1023/B:NEUR.0000044189.08240.a2
Kreshuk A, Straehle CN, Sommer C, Koethe U, Cantoni M, Knott G, Hamprecht FA (2011) Automated detection and segmentation of synaptic contacts in nearly isotropic serial electron microscopy images. PLoS One 6(10):e24899
Andres B, Köthe U, Helmstaedter M, Denk W, Hamprecht FA (2008) Segmentation of SBFSEM volume data of neural tissue by hierarchical classification. In: Rigoll G (ed) Pattern Recognition. LNCS, vol 5096. Springer, Berlin, Heidelberg, pp 142–152
Jain V, Murray J, Roth F, Turaga S, Zhigulin V, Briggman K, Helmstaedter M, Denk W, Seung H (2007) Supervised learning of image restoration with convolutional networks. IEEE 11th International Conference on Computer Vision, pp 1–8, Rio de Janeiro, Brazil, 14–21 October 2007
Jeong W-K, Beyer J, Hadwiger M, Blue R, Law C, Vazquez-Reina A, Reid RC, Lichtman J, Pfister H (2010) Secret and neurotrace: Interactive visualization and analysis tools for large-scale neuroscience data sets. IEEE Comput Graph 30(3):58–70
Jurrus E, Whitaker R, Jones B, Marc R, Tasdizen T (2008) An optimal-path approach for neural circuit reconstruction. In: Proceedings of the 5th IEEE International Symposium on Biomedical Imaging: From Nano to Macro, pp 1609–1612, Paris, France, 14–17 May 2008
Macke J, Maack N, Gupta R, Denk W, Schölkopf B, Borst A (2008) Contour-propagation algorithms for semi-automated reconstruction of neural processes. J Neurosci Meth 167:349–357
Allen BA, Levinthal C (1990) Cartos II semi-automated nerve tracing: Three-dimensional reconstruction from serial section micrographs. Comput Med Imag Graph 14(5):319–329
Jurrus E, Tasdizen T, Watanabe S, Davis MW, Jorgensen EM, Whitaker RT (2008) Semi-automated reconstruction of the neuromuscular junctions in the c. elegans. In: MICCAI Workshop on Microscopic Image Analysis with Applications in Biology, New York, NY, 5–6 September 2008
Jurrus E, Watanabe S, Paiva A, Ellisman M, Jorgensen E, Tasdizen T (2012) Semi-automated neuron boundary detection and slice traversal algorithm for segmentation of neurons from electron microscopy images. Neuroinformatics 11(1):5-29
Funke J, Andres B, Hamprecht FA, Cardona A, Cook M (2011) Multi-hypothesis crf-segmentation of neural tissue in anisotropic em volumes. CoRR abs/1109.2449
Anderson JR, Mohamed S, Grimm B, Jones BW, Koshevoy P, Tasdizen T, Whitaker R, Marc RE (2010) The viking viewer for connectomics: scalable multi-user annotation and summarization of large volume data sets. J Microsc 241(1):1328
Vazquez L, Sapiro G, Randall G (1998) Segmenting neurons in electronic microscopy via geometric tracing. In: Proceedings of International Conference on Image Processing, pp 814–818, San Diego, CA, 12–15 October 1998
Bertalmío M, Sapiro G, Randall G (2000) Morphing active contours. IEEE Trans Pattern Anal Mach Intell 22:733–737
Jurrus E, Hardy M, Tasdizen T, Fletcher P, Koshevoy P, Chien CB, Denk W, Whitaker R (2009) Axon tracking in serial block-face scanning electron microscopy. Med Image Anal 13:180–188
Reina AV, Miller E, Pfister H (2009) Multiphase geometric couplings for the segmentation of neural processes. In: Proceedings of IEEE Conference on Computer Vision and Pattern Recognition, pp 2020–2027, Miami, FL, 20–25 June 2009.
Jeong W-K, Beyer J, Hadwiger M, Vazquez A, Pfister H, Whitaker RT (2009) Scalable and interactive segmentation and visualization of neural processes in em datasets. IEEE Trans Visual Comput Graph 15(6):1505–1514
Vazquez-Reina A, Miller E, Pfister H (2009) Multiphase geometric couplings for the segmentation of neural processes. In: Proceedings of IEEE Conference on Computer Vision and Pattern Recognition, pp 2020–2027, Miami, FL, 20–25 June 2009
Tasdizen T, Whitaker RT, Marc RE, Jones BW (2005) Enhancement of cell boundaries in transmission electron microscopy images. In: Proceedings of International Conference on Image Processing, vol 2, pp 129–132, Genoa, Italy, 11–14 September 2005
Kumar R, Va andzquez Reina A, Pfister H (2010) Radon-like features and their application to connectomics. In: IEEE Computer Society Conference on CVPRW, pp 186–193, San Francisco, CA, 13–18 June 2010
Akselrod-Ballin A, Bock D, Reid RC, Warfield SK (2009) Improved registration for large electron microscopy images. In: Proceedings of IEEE International Symposium on Biomedical Imaging, pp 434–437, Boston, MA, 28 June–1 July 2009
Preibisch S, Saafeld S, Tomancak P (2009) Globally optimal stitching of tiled 3d microscopic image acquisitions. Bioinformatics 25(11):1463–1465
Vu N, Manjunath B (2008) Graph cut segmentation of neuronal structures from transmission electron micrographs. In: Image Processing, 2008. ICIP 2008. 15th IEEE International Conference on, pp 725–728, San Diego, CA, 12–15 October 2008
Yang H-F, Choe Y (2009) Cell tracking and segmentation in electron microscopy images using graph cuts. In: Proceedings of IEEE International Symposium on Biomedical Imaging: From Nano to Macro, Boston, MA, 28 June–1 July 2009
Yang H-F, Choe Y (2009) 3D volume extraction of densely packed cells in em data stack by forward and backward graph cuts. Computational Intelligence for Multimedia Signal and Vision Processing, pp 47–52, Nashville, Tennessee, 30 March–2 April 2009
Kaynig V, Fuchs T, Buhmann JM (2010) Neuron geometry extraction by perceptual grouping in ssTEM images. In: IEEE Conference on Computer Vision and Pattern Recognition, San Francisco, CA, 13–18 June 2010
Venkataraju KU, Paiva A, Jurrus E, Tasdizen T (2009) Automatic markup of neural cell membranes using boosted decision stumps. In: EEE International Symposium on Biomedical Imaging (ISBI): From Nano to Macro, Boston, MA, 28 June–1 July 2009
Jurrus E, Paiva ARC, Watanabe S, Anderson J, Whitaker BWJRT, Jorgensen EM, Marc R, Tasdizen T (2010) Detection of neuron membranes in electron microscopy images using a serial neural network architecture. Med Image Anal 14(6):770–783
Geman S, Geman D (1984) Stochastic relaxation, gibbs distributions and the bayesian restoration of images. IEEE Trans Pattern Anal Mach Intell 6:721–741
Freeman WT, Pasztor EC, Owen T, Carmichael Y (2000) Merl a mitsubishi electric research laboratory. Int J Comput Vis 40:2000
Lafferty J, McCallum A, Pereira F (2001) Conditional random fields: Probabilistic models for segmenting and labeling sequence data. In: International Conference on Machine Learning, pp 282–289, Williams College, Williamstown, MA, 28 June–1 July 2001
Kumar S, Hebert M (2003) Discriminative random fields: A discriminative framework for contextual interaction in classification. In: ICCV, pp 1150–1157, Nice, France, 14–17 October 2003
Jain V (2010) Machine Larning of Image Analysis with Convolutional Networks and Topological Constraints. PhD Thesis, MIT
Turaga SC, Briggman KL, Helmstaedter M, Denk W, Seung HS (2009) Maximin learning of image segmentation. In: Advances in Neural Information Processing Systems, Vancouver, British Columbia, Canada, 7–10 December 2009
Turaga SC, Murray JF, Jain V, Roth F, Helmstaedter M, Briggman KL, Denk W, Seung HS (2010) Convolutional networks can learn to generate affinity graphs for image segmentation. Neural Comput 22(2):511–538
Jain V, Bollmann B, Richardson M, Berger DR, Helmstaedter MN, Brigmann KL, Bowden JB, Mendenhall JM, Abraham WC, Harris KM, Kasthuri N, Hayworth KJ, Schalek R, Tapia JC, Lichtmann JW, Seung HS (2010) Boundary learning by optimization with topological constraints. In: IEEE Conference on Computer Vision and Pattern Recognition, San Francisco, CA, 13–18 June 2010
Veeraraghavan A, Genkin AV, Vitaladevuni S, Scheffer L, Xu S, Hess H, Fetter R, Cantoni M, Knott G, Chklovskii D (2010) Increasing depth resolution of electron microscopy of neural circuits using sparse tomographic reconstruction. In: IEEE Conference on Computer Vision and Pattern Recognition, San Francisco, CA, 13–18 June 2010
Fukushima K (1982) Neocognitron: A new algorithm for pattern recognition tolerant of deformations and shifts in position. Pattern Recogn 15(6):455–469
Hubel D, Wiesel T (1962) Receptive fields, binocular interaction and functional architecture in the cat’s visual cortex. J Physiol 160:106–154
LeCun Y, Bengio Y (1995) Convolutional networks for images, speech, and time series. The handbook of brain theory and neural networks. MIT Press, Cambridge, MA, pp 255–258
Garcia C, Delakis M (2004) Convolutional face finder: A neurl architecture for fast and robust face detection. IEEE Trans Pattern Anal Mach Intell 26(11):1408–1423
Osadchy R, Miller M, LeCun Y (2005) Synergistic face detection and pos estimation with energy-based model. In: Advances in Neural Information Processing Systems, Vancouver, British Columbia, Canada, 5–8 December 2005
Lawrence S, Giles CL, Tsoi AC, Back A (1997) Face recognition: A convolutional neural network approach. IEEE Trans Neural Network 8(1):98–113
LeCun Y, Huang FJ, Bottou L (2004) Learning methods for generic object recognition with invariance to pose and lighting. In: Proceedings of the 2004 I.E. Computer Society Conference on Computer Vision and Pattern Recognition, vol 2, pp II–97–104, Washington, DC, 27 June–2 July 2004
Huang FJ, LeCun Y (2006) Large-scale learning with svm and convolutional nets for generic object categorization. In: IEEE Conference on Computer Vision and Pattern Recognition, New York, NY, 17–22 June 2006
Ning F, Delhomme D, Lecun Y, Piano F, Bottou L, Barbano PE (2005) Toward automatic phenotyping of developing embryos from videos. IEEE Trans Image Process 14:1360–1371
Seyedhosseini M, Kumar R, Jurrus E, Guily R, Ellisman M, Pfister H, Tasdizen T (2011) Detection of neuron membranes in electron microscopy images using multi-scale context and radon-like features. In: Medical Image Computing and Computer-Assisted Intervention MICCAI 2011. Lecture Notes in Computer Science (LNCS), vol 6891. Springer, Berlin, Heidelberg, pp 670–677
Lee H, Grosse R, Ranganath R, Ng AY (2009) Convolutional deep belief networks for scalable unsupervised learning of hierarchical representations. In: International Conference on Machine Learning, vol. 382, pp 609–616, Montreal, Quebec, Canada, 2009
Norouzi M, Ranjbar M, Mori G (2009) Stacks of convolutional restricted boltzmann machines for shift-invariant feature learning. In: IEEE Conference on Computer Vision and Pattern Recognition, Miami, FL, 20–25 June 2009
Hinton GE, Osindero S (2006) A fast learning algorithm for deep belief nets. Neural Comput 18:2006
Ciresan D, Giusti A, Gambardella L, Schmidhuber J (2012) Neural networks for segmenting neuronal structures in EM stacks. In: ISBI Electron Microscopy Segmentation Challenge, Barcelona, Spain, 2–5 May 2012
Tu Z (2008) Auto-context and its application to high-level vision tasks. IEEE Conference on Computer Vision and Pattern Recognition, pp 1–8, Anchorage, Alaska, 24–26 June 2008
Tu Z, (2008) Auto-context and its application to high-level vision tasks. In: Proceedings of IEEE Computer Vision and Pattern Recognition, Anchorage, Alaska, 24–26 June 2008
Haykin S (1999) Neural networks: A comprehensive foundation, 2nd edn. Prentice-Hall, Upper Saddle River, NJ
Principe JC, Euliano NR, Lefebvre WC (2000) Neural and adaptive systems: Fundamentals through simulations. Wiley, New York
Pomerleau D (1993) Knowledge-based training of artificial neural networks for autonomous robot driving. In: Connell J, Mahadevan S (eds) Robot Learning. Kluwer Academic, Dordrecht, pp 19–43
Wells G, Venaille C, Torras C (1996) Promising research: Vision-based robot positioning using neural networks. Image Vis Comput 14:715–732
Cottrell G (1990) Extracting features from faces using compression networks: face, identity, emotion and gender recognition using holons. In: Connection models: Proceedings of the 1990 summer school. Morgan Kaufmann, San Mateo, CA, pp 328–337
Rabi G, Lu S (1998) Visual speech recognition by recurrent neural networks. J Electron Imag 7:61–69
Venkatataju KU, Paiva A, Jurrus E, Tasdizen T (2009) Automatic markup of neural cell membranes using boosted decision stumps. In: Proceedings of the 6th IEEE International Symposium on Biomedical Imaging, pp 1039–1042, Boston, MA, 28 June–1 July 2009.
Leung T, Malik J (2001) Representing and recognizing the visual appearance of materials using three-dimensional textons. Int J Comput Vision 43(1):29–44
Schmid C (2001) Constructing models for content-based image retrieval. In: Computer Vision and Pattern Recognition, 2001, CVPR 2001. Proceedings of the 2001 I.E. Computer Society Conference on, vol. 2, pp II–39–II–45, Kauai, HI, 8–14 December 2001
Varma M, Zisserman A (2003) Texture classification: are filter banks necessary? In: Computer Vision and Pattern Recognition, 2003. Proceedings of the 2003 I.E. Computer Society Conference on, vol. 2, pp II–691–8, Madison, WI, 16–22 June 2003
Awate SP, Tasdizen T, Whitaker RT (2006) Unsupervised Texture Segmentation with Nonparametric Neighborhood Statistics. In: Proceedings of the European Conference on Computer Vision. pp 494–507, Graz, Austria, 7–13 May 2006
Awate SP, Whitaker RT (2006) Unsupervised, information-theoretic, adaptive image filtering for image restoration. IEEE Trans Pattern Anal Mach Intell 28(3):364–376
Buades A, Coll B, Morel J-M (2005) A non-local algorithm for image denoising. In: Proceedings of IEEE Conference on Computer Vision and Pattern Recognition, pp 60–65, San Diego, CA, 20–26 June 2005
Tasdizen T (2008) Principal components for non-local means image denoising. In: Proceeding of International Conference on Image Processing, San Diego, California, 12–15 October 2008
Breiman L (2001) Random forests. Mach Learn 45(1):5–32
Jin Y, Hoskins R, Horvitz HR (1994) Control of type-D GABAergic neuron differentiation by C. elegans UNC-30 homeodomain protein. Nature 372:780–783
White JQ, Nicholas T, Gritton J, Truong L, Davidson ER, Jorgensen EM (2007) The sensory circuitry for sexual attraction in C. elegans males. Curr Biol 17:1847–1857
Cybenko G (1989) Approximation by superpositions of a sigmoidal function. Math Contr Signals Syst 2:303–314
Hornik K (1991) Approximation capabilities of multilayer feedforward networks. Neural Network 4(2):251–257
Cardona A, Saalfeld S, Schindelin J, Arganda-Carreras I, Preibisch S, Longair M, Tomancak P, Hartenstein V, Douglas RJ (2012) Trakem2 software for neural circuit reconstruction. PLoS One 7(6):e38011
Acknowledgments
This work was supported by NIH R01 EB005832 and 1R01NS075314. The C. elegans dataset was provided by the Jorgensen Lab at the University of Utah. The mouse neuropil dataset was provided by the National Center for Microscopy Imaging Research. The retina dataset was provided by the Marc Lab at the University of Utah. The drosophila VNC dataset was provided by the Cardona Lab at HHMI Janelia Farm.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this chapter
Cite this chapter
Tasdizen, T., Seyedhosseini, M., Liu, T., Jones, C., Jurrus, E. (2014). Image Segmentation for Connectomics Using Machine Learning. In: Suzuki, K. (eds) Computational Intelligence in Biomedical Imaging. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-7245-2_10
Download citation
DOI: https://doi.org/10.1007/978-1-4614-7245-2_10
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4614-7244-5
Online ISBN: 978-1-4614-7245-2
eBook Packages: EngineeringEngineering (R0)