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Although fluorescent proteins are ubiquitously used as genetic tracers and imaging agents, there is significant room for improvement. This chapter discusses how new improved fluorescent proteins can be designed. It focuses on the design of far-red and infrared fluorescent protein ...

The discovery of multicolored fluorescent proteins (FPs), in reef corals, that are close relatives of the green fluorescent protein (GFP) has led to what is now viewed as the second GFP revolution. Numerous GFP-type proteins, termed “reef FPs,” have been cloned from reef organisms and many posse ...

Live confocal microscopy of vital fluorescent markers, expressed in mouse embryonic tissues, is a powerful and exciting method to study mammalian embryonic development. This chapter discusses imaging approaches to visualize and characterize dynamic changes of the yolk-sac vas ...

Target-specific imaging probes represent a promising tool in the molecular imaging of human cancer. Fluorescently-labeled target-specific probes are useful in imaging cancers because of their ability to bind a target receptor with high sensitivity and specificity. The develop ...

Specifically, gene-encoded biological probes serve as stable and high-performance tools to visualize cellular fate in living animals. The rat, as with the mouse, has offered important animal models for biology and medical research, and has provided a wealth of physiological and pharma ...

Despite the sequencing of parasite genomes and development of DNA microarray technology, gene profiling of parasites remains a difficult task. For example, transcriptome analysis cannot currently be applied to the hepatic stages of the malaria parasite Plasmodium falciparum due ...

Human papilloma virus (HPV) infection is considered one of the main factors involved in the pathogenesis of endocervical adenocarcinoma. However, the cellular location of HPV in this type of tumor is controversial. We have developed a method to determine the presence of HPV type 16 in endocerv ...

The herpes simplex virus (HSV) frequently establishes a latent state in neurons, which can then be reactivated from the infected neurons. Quantifying the single-cell viral load is essential for understanding latency and reactivation of this virus. In this chapter the methods of laser cap ...

Behind the exponential expansion of the acquired immunodeficiency syndrome (AIDS) epidemic, there is a continuous and progressive molecular evolution of human immunodeficiency virus (HIV)-1. In this regard, the molecular analysis of viral strains infecting several anatomic c ...

Lysing of a complete biopsy sample results in a mixture of desired and undesired proteins, reflecting the originating cell types. Therefore microdissecting tissue material is mandatory prior to sample lysis and all downstream applications of protein analysis (proteomics). The two m ...

The ability to define protein profiles of normal and diseased cells is important in understanding cell function. Laser capture microdissection permits the isolation of specific cell types for subsequent molecular analysis. In this study we have established conditions for obtaini ...

The anterior pituitary gland consists of a heterogeneous population of various cell types. To study a single cell type with a homogeneous cell population, one can perform laser capture microdissection (LCM). Because different pituitary cells have unique immunophenotypic profile ...

Macrophage foam cells are critical mediators in atherosclerosis plaque development. A better understanding of the in vivo transcript profile of foam cells during the formation and progression of lesions may lead to novel therapeutic interventions. Toward this goal, we demonstrate ...

The kidney consists of many functional modules called nephrons. Each nephron has a tubular structure made up of several structurally and functionally distinct segments. The analysis of individual segments requires the use of microdissection techniques. We describe protocols th ...

Microdissection techniques have become an important tool to link histomorphology and pathophysiological events using modern methods of molecular biology. They allow isolation of cell clusters or even single cells precisely under optical control from complex tissue structu ...

Laser-assisted microdissection has enabled the collection of morphologically defined cell populations from a tissue section. The PALM� Robot MicroBeam laser microdissection system provides a robust system for the retrieval of specified cells (including single cells). Due to t ...

Laser microbeam microdissection (LMM) is an increasingly important histological technique for obtaining homogeneous cell populations and tissue components in order to analyze target-specific changes in genes, gene expression, and proteins. The quality of data obtained with ...

Biological tissues (in particular those affected by disease) are inherently complex mixtures of different cell types and matrices. This heterogeneity can complicate the interpretation of molecular biological studies performed on whole-tissue extracts if the precise cellu ...

Single palpable nodules of the thyroid gland are common in clinical practice; the majority of such lesions are benign. However, noninvasive thyroid nodules that exhibit borderline morphological signs of papillary cancer represent a diagnostic challenge. Rearrangements of the RET ...

Microsatellite analysis is a frequently used method for detection of chromosomal deletions by loss of heterozygosity studies and for detection of microsatellite instability. For reliable microsatellite analyses, a tumor cell content of at least 80% is required. Therefore, laser m ...