Introduction and Historical Perspective

Abstract

During our combined 45 plus years of operating confocal microscopes and managing core microscopy facilities, and through teaching our Basic Confocal Microscopy Workshop at several venues, we have found that students and technicians who are novice users of confocal microscopes are often instructed by their mentors to go to the confocal system and collect some images. Often the implied message is that it should be easy and quick since it is only a microscope. Unfortunately, all too often the advisor of the student or supervisor of the technician does not have a full understanding of the complexity of a confocal microscope. Unless these novice users are in a situation where others have the time and knowledge to properly train them, their initial efforts often amount to an exercise in futility because key parameters are not properly considered. This leads to specimens that are not prepared properly and a lack of understanding of how to operate the confocal microscope in a way that maintains the fidelity of the specimen information. In too many instances, this lack of user training is exacerbated further because there is little or no daily oversight of the setup and maintenance of the microscope. In this combined scenario, neither the experimental preparation nor the microscopes are capable of producing the highest-quality information. As with the first edition, the goal of this book is to provide beginning and intermediate users of confocal microscopes of all types the basic knowledge required to effectively utilize the full capability of confocal imaging to provide high-quality image data. Each chapter has been thoroughly revised and updated to reflect advances in the field. We have also added information on new detectors and emerging techniques such as super-resolution microscopy and specimen clearing, updated the software and operation chapter, and added a chapter on how to do co-localization fluorescence microscopy studies correctly.