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Of all scientific instruments, probably none has had more applications in the life sciences than the light microscope. In Light Microscopy: Methods and Protocols, expert researchers explore the basics and the latest advances in microscope instrumentation, sample preparation, and imaging techniques, all of which have been producing fundamental insights into the functions of cells and tissues. Chapters cover a variety of bright field and fluorescence microscopy-based approaches that are central to the study of a range of biological questions, providing information on how to prepare cells and tissues for microscopic investigations, covering detailed staining procedures, and exploring methods to analyze images and interpret the results accurately. Composed in the highly successful Methods in Molecular Biology(TM) series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Comprehensive and current, Light Microscopy: Methods and Protocols is an essential handbook for all researchers who are exploring the intriguing microscopic world of the cell.

Light Microscopy: Methods and Protocols

TABLE OF CONTENTS:

Table of Contents

Preface

Contributors

Part I. Bright-field Microscopy Applications

1. Glycol methacrylate embedding for improved morphological, morphometrical and immunohistochemical investigations under light microscopy: testes as a model
Hélio Chiarini-Garcia, Gleydes Gambogi Parreira and Fernanda Raddichi Almeida

2. Histological processing of teeth and periodontal tissues for light microscopy analysis.
Gerluza A. Borges Silva, Adriana Moreira and José Bento Alves

3. Large plant samples: how to process for 2-hydroxyethyl methacrylate embedding?
Élder Antônio Sousa Paiva, Sheila Zambello de Pinho and Denise Maria Trombert Oliveira

4. Image cytometry: nuclear and chromosomal DNA quantification
Carlos Roberto Carvalho, Wellington Ronildo Clarindo and Isabella Santiago de Abreu

5. Histological approaches to study tissue parasitism during Trypanosoma cruzi infection
Daniela L. Fabrino, Lívia Teixeira, Grazielle Ribeiro and Rossana C.N. Melo

6. Intravital microscopy to study leukocyte recruitment in vivo
Vanessa Pinho, Fernanda Matos Coelho, Gustavo Batista Menezes and Denise Carmona Cara


Part II. Fluorescence Microscopy Applications

7. Introduction to fluorescence microscopy
Ionita Ghiran

8. Using the fluorescent styryl dye FM1-43 to visualize synaptic vesicles exocytosis and
endocytosis in motor nerve terminals
Ernani Amaral, Silvia Guatimosim and Cristina Guatimosim

9. Imaging lipid bodies within leukocytes with different light microscopy techniques
Rossana C. N. Melo, Heloísa D´Ávila, Patrícia T. Bozza and Peter F. Weller

10. EicosaCell - An immunofluorescent-based assay to localize newly synthesized
eicosanoid lipid mediators at intracellular sites.
Christianne Bandeira-Melo, Peter F. Weller and Patricia T. Bozza

11. Nestin-driven green fluorescent protein as an imaging marker for nascent blood vessels in mouse models of cancer
Robert M. Hoffman

12. Imaging calcium sparks in cardiac myocytes
Silvia Guatimosim, Cristina Guatimosim and Long-Sheng Song

13. Light microscopy in aquatic ecology: methods for plankton communities studies
Maria Carolina S. Soares, Lúcia M. Lobao, Luciana O. Vidal, Natália P. Noyma, Nathan O. Barros, Fábio Roland

14. Fluorescence immunohistochemistry in combination with differential interference contrast microscopy for studies of semi-ultrathin specimens of epoxy resin-embedding samples.
Shin-ichi Iwasaki and Hidekazu Aoyagi