Cells synthesize and store within membranous sacs products such as hormones, growth factors, neurotransmitters, or digestive enzymes, for release on demand. As recently as just 15 years ago, it was believed that during cell secretion, membrane-bound secretory vesicles completely me ...

One of the challenging tasks in molecular cell biology is to identify and localize specific binding sites on biological samples with high spatial accuracy (in order of several nm). During the past 5 years, simultaneous topography and recognition imaging (TREC) has become a powerful AFM-bas ...

Nerve Morphometry is one tool employed in the clinical assessment of peripheral sural nerve pathological abnormalities. A new method is presented in this chapter incorporating an unbiased approach to quantitative sural nerve evaluation. Using conventional epoxy embedded nerv ...

This chapter deals with the stereological quantification of structural characteristics of the lung. The aim of design-based stereological methods is the unbiased and efficient estimation of structural features without making any assumptions on the underlying nature of the biol ...

The scientific community has become very concerned about inappropriate image manipulation. In journals that check figures after acceptance, 20–25% of the papers contained at least one figure that did not comply with the journal’s instructions to authors. The scientific press continu ...

Correlative light–electron microscopy (CLEM) is a very effective technique that combines live-cell imaging and immuno-electron microscopy for ultrastructural morphological characterization of dynamic intracellular organelles. The use of green fluorescent prote ...

Correlative microscopic approaches combine the advantages of both light and electron microscopy. Here we show a correlative approach that uses the photooxidation capacity of fluorescent dyes. Through illumination with high energetic light, the chromogen diaminobenzidine ...

Detailed insight into the fine structure and 3D-architecture of the complex and dynamic compartments of the endocytic system is essential for a morpho-functional analysis of retrograde traffic from the cell surface to different intracellular destinations. Here, we describe a cyto ...

Peroxisomes and mitochondria are essential subcellular organelles in mammals. Interestingly, recent studies have elucidated that these highly dynamic and plastic organelles exhibit a much closer interrelationship than previously assumed. Peroxisomes and mitochond ...

Autophagy is a bulk intracellular degradation process that is ubiquitous in eukaryotic cells and helps to recycle nutrients from catabolites by degrading proteins, lipids, and glycans, including organelles. Since autophagy has divergent physiological roles in cancer, infect ...

Immunogold labeling (IGL) technique has been utilized by many authors in combination with scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to obtain the identification/localization of receptors and antigens, both in cells and tissues. Environme ...

Environmental scanning electron microscopy (ESEM) (1) is an imaging technique which allows hydrated, insulating samples to be imaged under an electron beam. The resolution afforded by this technique is higher than conventional optical microscopy but lower than conventional scan ...

Cells store excess lipid as esters in the form of triglycerides and cholesterol esters. Most lipid esters are compartmentalized in globular structures called lipid droplets. Here we describe several methods of detecting lipid droplets by fluorescence microscopy. Lipid droplets c ...

Using an electron microscope’s scanning transmission mode (STEM) for collection of tomographic datasets is advantageous compared to bright field transmission electron microscopic (TEM). For image formation, inelastic scattering does not cause chromatic aberration, since ...

Epi-fluorescence microscopy is available in most life sciences research laboratories, and when optimized can be a central laboratory tool. In this chapter, the epi-fluorescence light path is introduced and the various components are discussed in detail. Recommendations are made for ...

MALDI (Matrix-Assisted Laser Desorption/Ionization) Imaging mass spectrometry is a powerful new method for analyzing the spatial distribution of molecules in tissues. Several different classes of cellular constituents such as proteins, peptides, lipids, and small molecul ...

Fluorescence microscopy has revolutionized the way live-cell imaging is achieved. At the same time, it is also potentially harmful to a living specimen. Therefore, the specimen must be monitored for viability and health before, during, and after imaging sessions. Methods for monitoring c ...

Confocal laser scanning microscopy is commonly used to visualize and quantify protein expression. Visualization of the expression of multiple proteins in the same region via multifluorescence allows for the analysis of differential protein expression. The defining step of mult ...

The measurement of colocalization requires images of two fluorophores that are aligned, with no cross talk, and that the intensities remain within the response range of the microscope. Quantitation depends upon differentiating between the presence and absence of fluorescence, and m ...

Real-time imaging coupled with a permeabilized cell system presents a very versatile platform to visualize the dynamic and intricate nature of nuclear envelope breakdown, one of the major morphological changes of mitosis. Here, we describe such a strategy in which the plasma membrane of ce ...