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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 18 (1991), S. 192-196 
    ISSN: 0741-0581
    Keywords: F1 mitochondrial ATPase ; Immunocytochemistry ; Mouse heart muscle cells ; Protein A-gold complexes ; Electron spectroscopic imaging (ESI) ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Immunocytochemistry was used to investigate the localization of F1 ATPase in mitochondria of cryosections of adult mouse heart muscle cells. The initial aldehyde fixation was the only denaturation step for antigens. The fine structure was preserved with contrast enhancement as the sections were maintained hydrated, with the advantage that the entire procedure is completed in one working day. The reaction was highly specific, and entire mitochondria were labeled with the Protein A-gold complex. A new analytical technique, electron spectroscopic imaging (ESI), contributed to a better visualization of the localization of the F1 factor.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 85-102 
    ISSN: 1059-910X
    Keywords: Cerebellum ; Mesodiencephalon ; Spiny appendages ; Glomerulus ; Gap junction ; Rebound activation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper reports on the detailed morphology of inferior olivary neurons in the cat following electrophysiological examination, intracellular injection with horseradish peroxidase, and gamma aminobutyric acid (GABA) immunocytochemistry.The activity of olivary cells was recorded intracellularly in vivo and their response to mesodiencephalic stimulation was tested. In a number of cases their response to stimulation of the contralateral superior cerebellar peduncle was also tested. Mesodiencephalic stimulation resulted in monosynaptic, and superior peduncle stimulation in disynaptic activation of cells in the medial accessory and principal olivary subdivisions. Rebound olivary activity was usually only found after mesodiencephalic stimulation.Light microscopic investigation of osmicated and Araldite embedded Vibratome sections was facilitated considerably when performing the osmication in a glucose solution. Peroxidase labeled olivary cells, like that earlier described for Golgi-impregnated material, possess a complex globular dendritic geometry. Especially, and unlike Golgi material, the abundance of exceptionally long and complex spiny appendages could be appreciated. The axons usually stemmed from first order dendrites and did not give rise to recurrent axon collaterals.The ultrastructural analysis of this material, mainly from serial sections, was combined with postembedding GABA immunohistochemistry. In this way, GABAergic as well as non-GABAergic profiles were studied in conjunction with HRP labeled cellular elements.The GABAergic terminals usually contained pleomorphic vesicles and made symmetrical synapses whereas non-GABAergic terminals nearly always formed asymmetrical synapses and contained round or oval vesicles. Most, if not all, HRP labeled spiny appendages were incorporated in glomeruli. A particular spiny appendage may contribute more than one spine head to a glomerular core, which, on average, consisted of spiny elements of six different neurons. A glomerular core is surrounded by approximately the same amounts of GABAergic and non-GABAergic boutons. Also, all spiny appendages, and most of their individual spine heads, are contacted by GABAergic as well as non-GABAergic boutons. Spiny appendages on the axon hillock may be incorporated in dendritic glomeruli, however, most synapses with the hillock were made by GABAergic boutons.The combined physiological and morphological observations imply that (1) the cerebellar nucleican excert an excitatory influence on inferior olivary neurons through a mesodiencephalic relay, (2) the GABAergic nucleo-olivary input seems to be capable of diminishing the oscillatory tendencies of olivary neurons, and (3) the mesodiencephalic (non-GABAergic) and cerebellar (GABAergic) input may subserve a timing function since these inputs systematically impinge upon the same olivary spines. © 1993 Wiley-Liss, Inc.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 169-170 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 400-411 
    ISSN: 1059-910X
    Keywords: Bronchiolar epithelium ; Lung development ; SP-A ; SP-B ; SP-C mRNA ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The morphologic and functional differentiation of the nonciliated columnar (Clara) cell, one of two secretory cell types in distalmost bronchioles in mammals, was studied in the mouse. Lungs from embryos (16-19 days of developmental age, dDA; birth on day 19), postnatal animals (5-20 days postnatally dPN), and adult animals were investigated by transmission electron microscopy, using standard staining procedures and immunogold (GAR-Au10) labeling for SP-A and Clara cell 10 KD antigen (CCA). At 16 dDa, all the columnar epithelial cells lining prospective distalmost bronchioles lacked distinctive features. By 17 dDa, some cells displayed a few cilia or apical dense granules. At 18 dDa, many nonciliated columnar cells had apical protrusions, as are seen in adult Clara cells. Apical concentrations of glycogen observed in nonciliated columnar cells perinatally were absent by 5 dPN, whereas apical dense granules became more abundant. Profiles of smooth and rough endoplasmic reticulum (ER), which had been randomly distributed, exhibited a selective, adult distribution at 20 dPN (apical vs. basal cytoplasmic domains). Labeling for SP-A and CCA, which was almost absent between 17 and 19 dDa, reached adult levels at the same time. The two proteins differed in distribution. SP-A predominated in adluminal cytoplasmic areas, where it was found over dense granules, vesicles, and multivesicular bodies; it was also present in bronchiolar lumens and intercellular spaces but not in rough ER or Golgi apparatus. In contrast, CCA showed a more uniform distribution; it was present over the same structures as SP-A and in the synthetic organelles. Ciliated columnar cells were virtually devoid of SP-A and CCA. We conclude that mouse Clara cells acquire a mature phenotype by 20 dPN. They are likely to be involved in recycling and/or degradation of SP-A that is internalized from airway lumens through their apical or lateral cell borders; furthermore, they synthesize the Clara cell 10 kD protein. These two Clara cell functions (first detectable late prenatally) reach mature levels by 20 dPN. © 1993 Wiley-Liss, Inc.
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  • 5
    ISSN: 1059-910X
    Keywords: Nitric oxide ; NADPH diaphorase ; Immunocytochemistry ; Retrograde tracing ; Prevertebral ganglion ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The distribution of neurons that are capable of synthesizing nitric oxide (NO) has been demonstrated in the porcine large intestine by means of NO synthase (NOS) immunocytochemistry and nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) histochemistry. An overall colocalization of NOS immunoreactivity and NADPHd staining was observed. Nitrergic neurons were abundant in the myenteric and outer submucous plexus of the caecum, colon, and rectum. Only a few nitrergic perikarya were seen in the inner submucous plexus of the colon and caecum, whereas a substantially larger number was observed in the rectum. Nitrergic nerve fibers were present in the three ganglionic nerve plexuses. Contrary to the outer longitudinal muscle layer and the mucosal region, the circular muscle layer received a dense nitrergic innervation. The nitrergic nerve cells were variable in size and shape, and several displayed vasoactive intestinal polypeptide (VIP) immunoreactivity (IR). Retrograde tracing studies revealed the existence of nitrergic neurons that project to the caudal (inferior) mesenteric ganglion. They were observed in the myenteric and outer submucous plexus of the transverse and descending colon and the rectum. These observations strongly suggest that several subpopulations of NO-synthesizing neurons, namely, motor neurons and interneurons, should be distinguished in the porcine large intestine, thereby emphasizing the importance of NO as a biologically active mediator. © 1994 Wiley-Liss, Inc.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. 103-111 
    ISSN: 1059-910X
    Keywords: Acetylcholinesterase ; NADHd ; NADPHd ; NOSynthase ; Enteric nervous system ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Using acetylcholinesterase (AChE), nicotinamide adenine dinucleotide diaphorase (NADHd), and nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) enzyme histochemical techniques, the ganglionated plexuses of the porcine enteric nervous system were investigated in small intestine whole-mount preparations. Both AchE and NADHd techniques revealed a majority of the neurons in the ganglia of all three major plexuses. The AchE technique also demonstrated clearly the axodendritic networks of the plexus myentericus. Intraganglionic blank areas revealed the localization of negative cell groups. A very high correlation was found between the activity of both enzymes in one neuron, although this correlation was certainly not linear. Many neurons exhibited a stronger signal for one enzyme. A very small part of the positive nerve cells showed intense staining for both AchE and NADHd. The NADPHd technique demonstrated that the NADPHd-positive neurons fill the negative intraganglionic spaces in the ganglia. Double staining with the two other enzymes showed virtually no colocalization of NADPHd with either NADHd or AchE in the porcine jejunal enteric ganglia. Little negative intraganglionic spaces were seldom found, leaving room for perhaps still more negative enteric neurons. Based upon these results we suggest that the enteric neurons of the porcine small intestine can be subdivided into AchE-NADHd and NADPHd subpopulations. Since the latter colocalizes with the neuronal NO synthase enzyme, we further suggest a subdivision of the enteric nerve cells into AchE-NADHd and NOS-NADHd neurons. © 1994 Wiley-Liss, Inc.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 268-273 
    ISSN: 1059-910X
    Keywords: Light ; Testis ; Ram ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This experiment was conducted in Ile-de-France adult rams to examine the target point of a 2-month light cycle regimen on seminiferous tubule functions, on intertubular compartment and on Leydig cell parameters. Eight rams were subjected to a 2-month light cycle regimen and were compared to sexually active or inactive rams. In light-treated rams, testis weight was maintained equal or was higher than that of sexually active rams. Both tubular and intertubular tissues were found significantly higher in light-treated than in sexually active rams. The mean ratio of basement membrane area of the seminiferous tubules per Sertoli cells and the daily productions of A1 spermatogonia and of leptotene primary spermatocytes were significantly increased in light-treated rams as compared with sexually active or inactive rams. Meanwhile, the dairy productions of diplotene primary spermatocytes, of round spermatids, of spermatozoa and of the rete testis fluid were not significantly increased in light-treated as compared with sexually active rams but significantly greater than those of sexually inactive rams. Total volume, total numbers, and individual volumes of Leydig cells were at least equal or higher in light-treated than in sexually active rams.
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  • 8
    ISSN: 1059-910X
    Keywords: Image analysis ; Respiratory cells ; SEM ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper describes the coupling between a scanning electron microscope (SEM) and an image analysis workstation. The system was designed in order to drive the SEM and to analyse any sample. It allows automatic (edge detection) or semiautomatic (pointing, marking, drawing) object detection. Two types of data can be obtained: (1) topographical information, such as the location of the object within a region of interest drawn at any magnification of the microscope, or (2) quantitative data, such as morphometric characteristics of objects. In addition, high resolution maps of the section, regions of interest, and objects can be obtained with a laser printer. This software was first applied to quantitate the adhesion of the bacteria Pseudomonas aeruginosa to human respiratory epithelial cells in culture. P. aeruginosa was shown associated with ciliated cells. The second application concerned the study of the distribution of specific carbohydrate residues at the surface of the respiratory cells. The gal residues were revealed using the lectin Ricinas communis agglutinin II, adsorbed to colloidal gold particles. A relationship between the presence of adherent bacteria and labelling was shown. © 1993 Wiley-Liss, Inc.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 362-373 
    ISSN: 1059-910X
    Keywords: ESEM ; Hydrophobic ; Hydrophilic ; Water condensation ; Water droplets ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: With the ability to perform dynamic experiments in the environmental scanning electron microscope (ESEM), the evaluation of microporous polymer membranes via a scanning electron microscope has advanced beyond morphological and elemental analysis. By adjusting sample temperature and environmental chamber pressure, the process of condensing water onto the porous membrane surface can be achieved. In doing so, assessments about the uniformity of wetting in hydrophilic membranes can be obtained based on how the liquid water spreads. Variations in the shape of condensed water droplets formed on non-water wetting structures will reflect the degree of hydrophobicity. This technique has proven useful in the characterization of hydrophobic spots on chemically modified hydrophilic structures and the dynamic examination of irregular wetting patterns in naturally hydrophilic membranes. © 1993 Wiley-Liss, Inc.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 28 (1994), S. 356-367 
    ISSN: 1059-910X
    Keywords: Hairy cell leukemia ; Immunogold labelling ; B-ly7 antibody ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Two cases of hairy cell leukemia have been studied by immuno-TEM and immuno-SEM after immunogold labelling of the cell surface antigen recognized by the B-ly7 monoclonal antibody.Most hairy cells appeared significantly labeled, although the density of the expression of the antigen, as demonstrated by immunogold labelling, seems variable from cell to cell. Moreover, some cells with the morphology of hairy cells and which could not be identified as monocytes were not labeled. Labelling for the antigen identified by the B-ly7 mAb does not seem to correlate with the presence of ribosome lamellae complexes which were present only in one of the two cases studied. Rare lymphocytes of unidentified lineage were labeled. Monocytes were significantly absent from the samples of peripheral blood of the two patients studied. In one normal control sample, monocytes were observed unlabelled.The results are discussed in reference to the pathogenesis of hairy cell leukemia, its surprisingly low mitotic rate, and its distinct response to chemotherapy. © 1994 Wiley-Liss, Inc.
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