Unbiased Stereology, Second Edition is a practical guide to making unbiased 3-D measurements via the microscope. Only those stereological techniques which have been tried and tested by real application are included. Although this technology is essentially mathematical and statistical, the authors do not immerse the reader in complex analysis, but rather provide simple heuristic explanations and references to the original proof, and illustrate the theory by analogies drawn from everyday experience. To give practical experience in application of the techniques, exercises are provided at the end of each chapter, complete with detailed worked answers.

Stereological techniques allow biologists to create quantitative, three-dimensional descriptions of biological structures from two- dimensional images of tissue viewed under the microscope. For example, they can accurately estimate the size of a particular organelle, the total length of a mass of capillaries, or the number of neurons or synapses in a particular region of the brain. This book provides a practical guide to designing and critically evaluating stereological studies of the nervous system and other tissues. It explains the basic concepts behind design-based stereology and how to get started. Also included are detailed descriptions of how to prepare tissue appropriately, perform pilot studies and decide on the appropriate sampling strategy, and account for phenomena such as tissue shrinkage. Numerous examples of applications of stereological methods that are applicable to studies of the central system and a wide variety of other tissues are explained. The book is therefore essential reading for neurobiologists and cell biologists interested in generating accurate representations of cell and tissue architecture.

vi on geometric probability is included, students can be expected to create a few simple programs like those shown, but for other geometries. I am indebted to Tom Hare for critical reviews of the material and an endless enthusiasm to debate and derive stereological relationships; to John Matzka at Plenum Press for patiently instructing me in the intricacies of typesetting; to Chris Russ for helping to program many of these measurement techniques; and especially to Helen Adams, both for her patience with my creative fever to write yet another book, and for pointing out that the title, which I had intended to contrast to "theoretical stereology," can also be understood as the antonym of "impractical stereology." John C. Russ Raleigh,NC July, 1986 Chapter 1: Statistics 1 Accuracy and precision 1 The mean and standard deviation 5 Distributions 7 Comparison 13 Correlation 18 Nonlinear fitting 19 Chapter 2: Image Types 23 Planar sections 23 Projected images 25 Finite sections 28 Space-filling structures and dispersed phases 29 Types of images and contrast mechanisms 31 Sampling 32 Chapter 3: Manual Methods 35 Volume fraction 35 Surface density 38 Contiguity 41 Mean intercept length 42 Line density 43 Grain size determination 55 Curvature 48 Reticles to aid counting 49 Magnification and units 51 Chapter4: Size Distributions 53 Intercept length in spheres 53 Nonspherical shapes 57 Corrections for finite section thickness 59 Lamellae 61 Measurement of profile size 62 Nonspherical particles 69 vii Contents viii Chapter 5: Computer Metlwds 73

This update to Peter R. Mouton's pioneering work provides bioscientists with the concepts needed in order to apply the principles and practices of unbiased stereology to research involving biological tissues. Mouton starts with a brief explanation of the history and theory of the process before defining the terms, concepts, and tools of unbiased stereological procedures. He compares and contrasts the procedures with less-exacting approaches to quantitative analysis of biological structure using specific examples from biomedical literature. The book incorporates existing best practices with new methodologies, such as the Rare Event Protocol, while simplifying the dense, often difficult literature on the Subject to show the utility and importance of unbiased stereology. This clear, insightful guide goes a step further than other books on this Subject by demonstrating not only how to use unbiased stereology but also how to interpret and present the results. Written by the official U.S. representative to the International Society for Stereology, this is the most complete, up-to-date resource on the science of unbiased stereology. Those new to bioscience research as well as experienced practitioners will find that Mouton's explanations are the perfect companion for stereology courses and workshops.

Stereological methods provide researchers with unparalleled quantitative data from tissue samples and allow for well-evidenced research advances in a broad range of scientific fields. Presenting a concise introduction to the methodology and application of stereological research in neuroscience, Neurostereology provides a fuller understanding of the use of these methods in research and a means for replicating successful scientific approaches. Providing sound footing for future research, Neurostereology is a useful tool for basic and clinical researchers and advanced students looking to integrate these methods into their research.

Over the past few decades an exceedingly large number of experimental and clinical investigations have been performed in an attempt to analyze the way in which the kidney functions. The basis for all this work was established during the nineteenth and the early twentieth century by morphologists (Bowman 1842; Hyrtl1863, 1872; Heidenhain 1874; Peter 1909; von Mollendorf 1930). All these investigators clearly outlined the extremely heterogeneous assembly of renal tissue and also defined the nephron as the smallest morphological unit. It was further the merit of these anato mists and histologists to preclude quite a number of nephron functions based merely on their careful observations. Contemporary histologists have been able to add little to these observations. Unfortunately with the introduction of physiologic in vivo et situ studies on kidneys the interest in heterogeneity waned. This lack of attention was aggravated by the introduction of the clearance techniques which cannot account for regional differences in the function of the smallest unit, the nephron. That ana tomic heterogeneity has a functional correlate was strongly suggested by Trueta et al. (1947) and vigorously stimulated a number of studies. The development of physiologic microtechniques, like micropuncture and microperfusion of single nephrons, or the perfusion of isolated nephron portions and electrophysiologic studies, enormously expanded our knowledge concerning details regarding nephron and total renal func tion.