Molecular Cytogenetics: Protocols and Applications

Abstract

In 1986, Pinkel and Gray first utilized FISH techniques in cytogenetic diagnoses. FISH technology, a combination of molecular and conventional cytogenetic techniques, has brought modern cytogenetics to a new era with significantly higher resolutions and a much wider testing spectrum. Since then, numerous new FISH-based technologies have been emerging, from metaphase FISH to interphase FISH and then fiber FISH, from single-color FISH to spectral karyotyping (SKY) and multicolor FISH (M-FISH), from regular FISH to reverse FISH (also called “microFISH”), from comparative genomic hybridization (CGH) to microarray CGH, and so on. Each technique is associated with a specific testing spectrum and limitations. It is neither easy for clinicians to choose the right tests for their patients—i.e., those that will bring the benefits afforded by the new technologies—nor straightforward for clinical cytogeneticists to decide which new technologies to implement in their laboratories with limited laboratory budgets and low reimbursement rates.Although many books and articles about FISH technologies and their clinical applications have been published, a book that contains step-by-step and easy-to-follow protocols for most of the new FISH technologies was not available until the recent publication of Molecular Cytogenetics: Protocols and Applications, edited by Yao-Shan Fan. Dr. Fan has done a marvelous job of putting together a collection of laboratory protocols and review articles of different FISH technologies. The book is divided into four parts: (1) “Basic Concepts and Techniques,” including FISH probe sources, fluorophores, probe labeling, and basic FISH procedures; (2) “Evolving Techniques and Applications,” including chromosome microdissection (microFISH), primed in situ labeling (PRINS), SKY, M-FISH, CGH, color banding, fiber FISH, telomere FISH, and microarray CGH; (3) “Special Applications in Chromosomal Disorders,” including delineating marker chromosomes, prenatal diagnosis of common aneuploidies, prenatal diagnosis using fetal cells in maternal blood, preimplantation test, microdeletion syndromes, uniparental disomy, and reproductive pathology; and (4) “Special Application in Oncology,” including FISH in chronic myeloid leukemia (CML), FISH detecting Her-2/neu amplification in breast cancer, CGH in cancer study, etc.The book focuses mainly on laboratory diagnosis. Most of the chapters were written in a format similar to a laboratory procedure manual, with step-by-step instructions. Some chapters are especially well written and deserve special mention: chapters 7, 8, 9, 12, and 18, describing SKY, M-FISH, CGH, multitelomere FISH, and preimplantation tests, respectively, are of exceptional value for clinical cytogenetic laboratory directors and technologists. The authors of the different chapters not only gave detailed, easy-to-follow protocols, but also provided their first-hand experiences of troubleshooting. Chapter 19 concisely summarizes common microdeletion and microduplication syndromes, a good reference for neonatologists and dysmorphologists. Although the book targets diagnosis, some chapters can be useful for researchers in the fields of human development and cancer biology, such as microdissection and microarray CGH.The most apparent drawback of this book is that it does not emphasize enough FISH applications in cancer genetics. According to the recently published The World Health Organization Classification of Neoplasms, one of the most important biological features used for the classification of cancer is the cytogenetic alteration of tumor cells. These alterations are directly associated with patient responses to therapies and are therefore of not only diagnostic but also prognostic significance. In many cytogenetic laboratories, FISH has become a major diagnostic method for hematopoietic malignancies and solid tumors. In this regard, Introduction to Fluorescence In Situ Hybridization, Principles and Clinical Applications, by Andreeff and Pinkel, would be an excellent reference for cytogeneticists as well as hematologists and oncologists, although it does not include step-by-step protocols, as does Dr. Fan’s book.We all understand that there is no book that includes everything about molecular cytogenetics. Compared with other related books, Molecular Cytogenetics: Protocols and Applications is packed with information on cutting-edge FISH technologies and their applications, limitations, and pitfalls, along with detailed laboratory protocols. I would certainly recommend that this book be included in the libraries of cytogeneticists and cytogenetic laboratories.