Dr. Pedro Barbosa is a Professor Emeritus at the Department of Entomology of the University of Maryland, College Park. He has been a professor at Rutgers University, the University of Massachusetts at Amherst and at the University of Maryland. Although his research focused on the ecology of insects and insect-plant interactions, he also has an interest in insect histology. Since his retirement in 2010 he has written several books on insects. Dr. Deborah L. Berry is Co-Director for the Histopathology and Tissue Shared Resource at the Lombardi Comprehensive Cancer Center, Georgetown University, which provides research pathology support for translational cancer research. She has over 20 years’ experience in histology with a specialty in the histology of the fruit fly Drosophila melanogaster. Dr. Christina S. Kary is a scientific editor with the journal Genes & Development at Cold Spring Harbor Laboratory Press, New York. Before becoming an editor, she earned a PhD with Dr. Eric Baehrecke at the University of Maryland and was a Jane Coffin Childs post-doctoral fellow with Dr. Susan Mango at Harvard University.

“A very useful book to have in your library.”  (British Journal of Entomology & Natural History, 1 March 2015)

• Preface ix Acknowledgements xiIntroduction xiiiAbout the companion website xiv1 Problems of sclerotized chitin: Softening insect cuticle 11.1 Introduction 11.2 General Methods 31.3 Preparations of insect eggs 141.4 Double Embedding Techniques 16References 192 Fixation 212.1 Introduction 212.2 Aldehyde based fixatives 212.3 Protein denaturing 302.4 Picric acid based 332.5 Mercuric chloride based 372.6 SEM/TEM 402.7 Other 46References 513 Dehydrating, clearing, and embedding 543.1 Dehydration 543.2 Clearing 603.3 Embedding General 653.4 Embedding – Ester Wax 733.5 Embedding – Methacrylate 74References 774 Staining 794.1 Single-contrast staining – Carmines 814.2 Single contrast staining – Nuclear Stains 834.3 Single contrast staining – General Stains 864.4 Single contrast staining – Golgi 894.5 Single contrast staining – Eggs 894.6 Single contrast staining – Silver Stains 904.7 Polychrome staining techniques – General 924.8 Polychrome staining – Brain/Nerve 1024.9 Polychrome staining – blood 1034.10 Single contrast procedures for chitinous material 1054.11 Polychrome staining procedures for chitinous material 1064.12 Polychrome staining for chitinous material – KOH 1104.13 Polychrome staining for chitinous material – Differential staining of Individual Organs 1114.14 Staining of specific tissues 1134.15 Two dye combinations 114References 1175 Immunohistochemical techniques 1195.1 Introduction 1195.2 General immunostaining techniques 1275.3 Immunolabeling of samples for Transmission Electron Microscopy (TEM) 1355.4 Proliferation assays 1405.5 Methods to detect specific proteins 142References 1446 Use of genetic markers in insect histology 1466.1 Introduction 1466.2 Inducible genetic markers 1496.3 Mosaic gene expression 1566.4 Fluorescent markers for live imaging and kinetic microscopy 165References 1697 Fluorescence 1717.1 Introduction 171References 1928 Mounting 1948.1 Introduction 194References 2069 Preparation of whole mounts 2089.1 Introduction 208References 22910 Preparation of whole mounts for staining 23110.1 Introduction 23110.2 Detection of NAPDHd 23710.3 SEM 23810.4 In situ hybridization 240References 24411 Preparation of genitalia, mouthparts and other body parts 246References 25612 Preparation of chromosomes 258References 28813 Preparation of other specific insect organs and tissues 29013.1 Introduction 290References 323Appendix Dissecting fluids and saline solutions 325Index 333

Dr. Pedro Barbosa is a Professor Emeritus at the Department of Entomology of the University of Maryland, College Park. He has been a professor at Rutgers University, the University of Massachusetts at Amherst and at the University of Maryland. Although his research focused on the ecology of insects and insect-plant interactions, he also has an interest in insect histology. Since his retirement in 2010 he has written several books on insects. Dr. Deborah L. Berry is Co-Director for the Histopathology and Tissue Shared Resource at the Lombardi Comprehensive Cancer Center, Georgetown University, which provides research pathology support for translational cancer research. She has over 20 years’ experience in histology with a specialty in the histology of the fruit fly Drosophila melanogaster. Dr. Christina S. Kary is a scientific editor with the journal Genes & Development at Cold Spring Harbor Laboratory Press, New York. Before becoming an editor, she earned a PhD with Dr. Eric Baehrecke at the University of Maryland and was a Jane Coffin Childs post-doctoral fellow with Dr. Susan Mango at Harvard University.

“A very useful book to have in your library.”  (British Journal of Entomology & Natural History, 1 March 2015)

• Preface ix Acknowledgements xiIntroduction xiiiAbout the companion website xiv1 Problems of sclerotized chitin: Softening insect cuticle 11.1 Introduction 11.2 General Methods 31.3 Preparations of insect eggs 141.4 Double Embedding Techniques 16References 192 Fixation 212.1 Introduction 212.2 Aldehyde based fixatives 212.3 Protein denaturing 302.4 Picric acid based 332.5 Mercuric chloride based 372.6 SEM/TEM 402.7 Other 46References 513 Dehydrating, clearing, and embedding 543.1 Dehydration 543.2 Clearing 603.3 Embedding General 653.4 Embedding – Ester Wax 733.5 Embedding – Methacrylate 74References 774 Staining 794.1 Single-contrast staining – Carmines 814.2 Single contrast staining – Nuclear Stains 834.3 Single contrast staining – General Stains 864.4 Single contrast staining – Golgi 894.5 Single contrast staining – Eggs 894.6 Single contrast staining – Silver Stains 904.7 Polychrome staining techniques – General 924.8 Polychrome staining – Brain/Nerve 1024.9 Polychrome staining – blood 1034.10 Single contrast procedures for chitinous material 1054.11 Polychrome staining procedures for chitinous material 1064.12 Polychrome staining for chitinous material – KOH 1104.13 Polychrome staining for chitinous material – Differential staining of Individual Organs 1114.14 Staining of specific tissues 1134.15 Two dye combinations 114References 1175 Immunohistochemical techniques 1195.1 Introduction 1195.2 General immunostaining techniques 1275.3 Immunolabeling of samples for Transmission Electron Microscopy (TEM) 1355.4 Proliferation assays 1405.5 Methods to detect specific proteins 142References 1446 Use of genetic markers in insect histology 1466.1 Introduction 1466.2 Inducible genetic markers 1496.3 Mosaic gene expression 1566.4 Fluorescent markers for live imaging and kinetic microscopy 165References 1697 Fluorescence 1717.1 Introduction 171References 1928 Mounting 1948.1 Introduction 194References 2069 Preparation of whole mounts 2089.1 Introduction 208References 22910 Preparation of whole mounts for staining 23110.1 Introduction 23110.2 Detection of NAPDHd 23710.3 SEM 23810.4 In situ hybridization 240References 24411 Preparation of genitalia, mouthparts and other body parts 246References 25612 Preparation of chromosomes 258References 28813 Preparation of other specific insect organs and tissues 29013.1 Introduction 290References 323Appendix Dissecting fluids and saline solutions 325Index 333