Joanne Berger-Sweeney

The Nissl Stain: A Stain for Cell Bodies in Brain Sections

INTRODUCTIONThe Nissl stain is used widely in many research labs to examine the overall morphology of the brain. It is often used to verify the location of a lesion or an electrode.

Dissection of Rat Brains

INTRODUCTIONIn this protocol, the perfused brain of a rat is separated from the surrounding tissue and post-fixed in a formalin/sucrose solution in preparation for freezing and sectioning. Dissection should be done as soon as possible after perfusion to prevent desiccation of the brain. This procedure can also be used to dissect fresh (non-perfused) brains, for example, for Golgi-Cox stain or neurochemical assays.

Sectioning of brain tissues.

INTRODUCTIONA fixed brain, although harder than the brain in its natural state, is still not hard enough to cut into very thin slices. To make uniform, thin sections, the brain must be frozen. A wide variety of sectioning devices are available, each with their own requirements. With a microtome, for example, a very sharp knife is drawn manually across a brain that has been frozen to a chilled platform using dry ice. After each cut, the knife is lowered mechanically by a preset amount, giving consistent sections of a required thickness. A drawback of this instrument is that the temperature of the platform can fluctuate, thus causing changes in the quality of the sections. Other instruments, such as a cryostat, maintain the platform-mounted brain and the knife inside an enclosed refrigerated compartment. The temperature is kept at -20°C, and electronics are used to control the speed of the knife and the thickness of the sections. Some models can be set up to make a series of sections automatically.

Observational methods used to assess rat behavior: general activity.

INTRODUCTIONThe activity-inactivity continuum is an important parameter of behavior, and quantification of overall locomotor activity in the rat should identify it as a naturally nocturnal animal. Disruptions in nocturnal activity can be caused by damage in visual inputs to the brain or damage in the hypothalamus. Many commercial devices are available to measure activity automatically; some can be integrated with a computer to allow overnight monitoring in the absence of an observer. A less sophisticated but still accurate method of measuring activity is to create a home-made activity chamber by replacing the bottom of a box with Plexiglas or by marking lines on the bottom of a clean rat cage so that the observer can record rat activity by noting when the lines are crossed, while simultaneously recording other behaviors. Activity in rat pups can be observed as soon as they are 10 days old using smaller activity chambers. This protocol describes the construction of a home-made activity chamber and how to measure four activities: locomotion, rearing, circling, and grooming.

Testing spatial and nonspatial learning using a morris water maze.

INTRODUCTIONAnimal studies of memory have used many different types of mazes and tasks, where the animal is required to learn the demands of the test and then perform correctly until a predetermined performance criterion is reached. Traditional studies have relied on water or food deprivation to motivate the animal to do the test using food or water rewards. In 1984, Richard Morris introduced the water maze. The maze consists of a large cylindrical tank of water with a hidden platform; the animal has to swim until it finds the platform. The animal generally uses cues outside the maze (extramaze cues) to develop a spatial map of the environment and guide its performance. This maze avoids the problems associated with food or water deprivation, because the animals goal is to find the hidden platform so that it can stop swimming. Many types of memory can be studied by varying the visual cues, the placement of the platform, and the starting point of the animal. This protocol describes the use of the water maze to test rats in two behavioral tasks: a spatial task and a nonspatial, visual task.

Choline Acetyltransferase Staining in Brain Sections

Cold Spring Harb Protoc; Carol Ann Paul, Barbara Beltz and Joanne Berger-Sweeney Choline Acetyltransferase Staining in Brain Sections Service Email Alerting click here. Receive free email alerts when new articles cite this article Categories Subject Cold Spring Harbor Protocols. Browse articles on similar topics from (90 articles) Visualization of Proteins (465 articles) Visualization (492 articles) Proteins and Proteomics, general (259 articles) Neuroscience, general (56 articles) Light Microscopy (303 articles) Labeling for Imaging (62 articles) Immunostaining Tissues (95 articles) Immunostaining (75 articles) Immunohistochemistry (543 articles) Imaging/Microscopy, general (277 articles) Imaging for Neuroscience (172 articles) Characterization of Proteins (1068 articles) Cell Biology, general (214 articles) Antibodies, general

Observational methods used to assess rat behavior: behavioral patterns.

INTRODUCTIONThis protocol describes a method for quantifying observation of normal rat behaviors in a controlled environment and in a defined area. It is necessary to have precise definitions of the behaviors to be quantified and a reliable procedure for quantifying them. The observation room should be quiet, and it should be equipped for light or dark observations (rats cannot see red light so dark observations can be made using a red light). Ideally, the observer should be separated from the animal by a one-way screen. If such a screen is not available, the observer should stand or sit in a constant, unobtrusive position and make as few movements as possible.