Springer protocols feed by Plant Sciences 
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This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Selected Simple Methods of Plant Cell Wall Histochemistry and Staining for Light Microscopy
Histochemical methods allow for identification and localization of various components within the tissue. Such information on the spatial heterogeneity is not available with biochemical methods. However, there is limitation of the specificity of such detection in context of complex tissue, which is important to consider, and interpretations of the results should regard suitable control treatments if possible. Hereby we present set of selected simple staining and histochemical methods with comments based on our laboratory experience. (Source: Springer protocols feed by Plant Sciences)
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Heterologous Expression in Budding Yeast as a Tool for Studying the Plant Cell Morphogenesis Machinery
The budding yeast (Saccharomyces cerevisiae) can serve as a unique experimental system for functional studies of heterologous genes, allowing not only complementation of readily available yeast mutations but also generation of overexpression phenotypes and in some cases also rescue of such phenotypes. Here we summarize the main considerations that have to be taken into account when using the yeast expression system for investigating the function of plant genes participating in cell morphogenesis; outline the strategies of experiment planning, yeast strain selection (or construction), and expression vector choice; and provi...
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Optical Trapping in Plant Cells
Optical tweezers allow noninvasive manipulation of subcellular compartments to study their physical interactions and attachments. By measuring (delay of) displacements, (semi-)quantitative force measurements within a living cell can be performed. In this chapter, we provide practical tips for setting up such experiments paying special attention to the technical considerations for integrating optical tweezers into a confocal microscope. Next, we describe some working protocols to trap intracellular structures in plant cells. (Source: Springer protocols feed by Plant Sciences)
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Laser Microdissection of Plant Cells
Different plant cell types express unique transcriptomes, proteomes, and metabolomes. Therefore, the isolation of specific cell types prior to molecular analyses is important to understand the specification, differentiation, and function of these cells. Isolation of specific plant cell types from composite organs can be achieved by laser microdissection (LMD). A wide variety of methods to fix and embed tissues prior to LMD and downstream molecular analyses have been developed for different plant species and tissues. The present review summarizes and highlights the most recently applied LMD approaches in plant science. (Sou...
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Lab-on-a-Chip for Studying Growing Pollen Tubes
A major limitation in the study of pollen tube growth has been the difficulty in providing an in vitro testing microenvironment that physically resembles the in vivo conditions. Here we describe the development of a lab-on-a-chip (LOC) for the manipulation and experimental testing of individual pollen tubes. The design was specifically tailored to pollen tubes from Camellia japonica, but it can be easily adapted for any other species. The platform is fabricated from polydimethylsiloxane (PDMS) using a silicon/SU-8 mold and makes use of microfluidics to distribute pollen grains to serially arranged microchannels. The tubes ...
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Essential Methods of Plant Sample Preparation for Light Microscopy
There are various preparatory techniques for light microscopy permitting access to the inner structure of plant body and its development. Minute objects might be processed as whole-mount preparations, while voluminous ones should be separated into smaller pieces. Hereby we summarize some of the “classical” techniques to cut more voluminous objects into slices and access their inner structure either for simple anatomical analysis or for further processing (e.g., histochemistry, immunohistochemistry, in situ hybridization, enzyme histochemistry). (Source: Springer protocols feed by Plant Sciences)
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Antisense Oligodeoxynucleotide-Mediated Gene Knockdown in Pollen Tubes
Specific gene knockdown mediated by the antisense oligodeoxynucleotides (AODNs) strategy recently emerged as a rapid and effective tool for probing gene role in plant cells, particularly tip-growing pollen tubes. Here, we describe the protocol for the successful employment of AODN technique in growing tobacco pollen tubes, covering AODN design, application, and analysis of the results. We also discuss the advantages and drawbacks of this method. (Source: Springer protocols feed by Plant Sciences)
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Plant Cell Lines in Cell Morphogenesis Research
Plant organs and tissues consist of many various cell types, often in different phases of their development. Such complex structures do not allow direct studies on behavior of individual cells. In contrast, populations of in vitro-cultured plant cells represent valuable tool for studying processes on a single-cell level, including cell morphogenesis. Here we describe characteristics of well-established model tobacco and Arabidopsis cell lines and provide detailed protocol on their cultivation, characterization, and genetic transformation. (Source: Springer protocols feed by Plant Sciences)
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Morphological Analysis of Cell Growth Mutants in Physcomitrella
This protocol describes a quantitative analysis of the morphology of small plants from the moss Physcomitrella patens. The protocol can be used for the analysis of growth phenotypes produced by transient RNA interference or for the analysis of stable mutant plants. Information is presented to guide the investigator in the choice of vectors and basic conditions to perform transient RNA interference in moss. Detailed directions and examples for fluorescence image acquisition of small regenerating moss plants are provided. Instructions for the use of an ImageJ-based macro for quantitative morphological analysis of these plant...
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Live Cell Imaging of Arabidopsis Root Hairs
Root hairs are tubular extensions from the root surface that expand by tip growth. This highly focused type of cell expansion, combined with position of root hairs on the surface of the root, makes them ideal cells for microscopic observation. This chapter describes the method that is routinely used in our laboratory for live cell imaging of Arabidopsis root hairs. (Source: Springer protocols feed by Plant Sciences)
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Immunogold Labeling of Resin-Embedded Electron Microscopical Sections
Knowledge about the spatio-temporal distribution patterns of proteins and other molecules of the cell is essential for understanding their function. A widely used technique is immunolabeling which uses specific antibodies to reveal the distribution of molecular components at various structural levels. Electron microscopy offers the highest resolution of morphological techniques and is thus an indispensable tool for the analysis of molecule distribution patterns at the subcellular level. In this chapter we describe a routine method for labeling ultrathin sections of resin-embedded material with antibodies conjugated to coll...
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Imaging Plant Nuclei and Membrane-Associated Cytoskeleton by Field Emission Scanning Electron Microscopy
Scanning electron microscopy (SEM) is a powerful technique that can image exposed surfaces in 3D. Modern scanning electron microscopes, with field emission electron sources and in-lens specimen chambers, achieve resolutions of better than 0.5 nm and thus offer views of ultrastructural details of subcellular structures or even macromolecular complexes. Obtaining a reliable image is, however, dependent on sample preparation methods that robustly but accurately preserve biological structures. In plants, exposing the object of interest may be difficult due to the existence of a cell wall. This protocol shows how to isolate pla...
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Reconstructing Plant Cells in 3D by Serial Section Electron Tomography
In micrographs acquired with a transmission electron microscope, 3-dimensional (3D) objects are superimposed onto a 2D screen. This reduction in dimension necessarily leads to a degradation of image resolution. To overcome this problem, 3D microscopy techniques, such as tomography and single particle analysis, have been developed. Tomography has been used to visualize cells in 3D, and single particle analysis has been used to investigate macromolecules and viral particles. In this chapter we will describe how we have collected tilting series micrographs from plant cells and how we have reconstructed the cellular volumes us...
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
High-Pressure Freezing and Low-Temperature Processing of Plant Tissue Samples for Electron Microscopy
Use of electron tomography methods improves image resolution of transmission electron microscopy especially in the z-direction, enabling determination of complicated 3D structures of organelles and cytoskeleton arrays. The increase in resolution necessitates preservation of cellular structures close to the native states with minimum artifacts. High-pressure freezing (HPF) that immobilizes molecules in the cell instantaneously has been used to avoid damages caused by convention chemical fixation. Despite the advantages of HPF, cells could still be damaged during dissection prior to HPF. Therefore, it is critical to isolate ...
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
Mechanical Measurements on Living Plant Cells by Micro-indentation with Cellular Force Microscopy
Indentation methods on the micro- and nanoscale are increasingly used to assess mechanical properties of living plant tissues. These techniques rely on recording the force resulting from indenting the cell surface with a small probe. Depending on the scale of indentation and the indenter shape, force-indentation data will reflect several factors such as cell wall elasticity, turgor pressure, cell and tip geometry, and contact angle. Cellular force microscopy is a micro-indentation method that was designed to precisely measure and apply forces on living plant cells. Here we explain how to use this method to map the apparent...
Source: Springer protocols feed by Plant Sciences - October 19, 2013 Category: Biology Source Type: news
