Microscopic and cytochemical characterisation of haemocytes of the mud crab Scylla serrata (Forskål, 1775) (Decapoda, Portunidae)

Crustaceana ◽  
2013 ◽  
Vol 86 (10) ◽  
pp. 1234-1249 ◽  
Author(s):  
Brajendu Kumar ◽  
A. Deepika ◽  
M. Arumugam ◽  
P. Mullainadhan ◽  
M. Makesh ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Toluidine Blue ◽  
Light And Electron Microscopy ◽  
Morphological Features ◽  
Scylla Serrata ◽  
Mud Crab ◽  
Cytoplasmic Organelles ◽  
Well Differentiated ◽  
Total Haemocyte Count ◽  
Prophenol Oxidase

Haemocytes of the mud crab Scylla serrata (Forskål, 1775) were characterised based on morphological features using light and electron microscopy, and cytochemistry. The cells were identified as hyaline, semigranular and granular haemocytes. Hyaline cells were the smallest haemocytes among the three types identified, having the highest nucleo-cytoplasmic ratio. The cells showed a number of cytoplasmic organelles and also contained a few small as well as large-sized granules. Semigranular haemocytes possessed moderate numbers of large-sized granules or numerous small-sized granules and comparatively less numbers of organelles. Granular haemocytes were the largest haemocytes with the lowest nucleo-cytoplasmic ratio and contained many large-sized granules. Cytoplasmic organelles were least observed in the granular haemocytes. These three haemocyte morphotypes constituted 60, 21 and 19%, respectively, of the total haemocyte population, while the total haemocyte count was 7.31 × 106 to 7.18 × 107 with a mean of 2.86 × 107 cells ml−1. In cytochemical studies performed to localize carbohydrates, lipids and prophenol oxidase, all the haemocyte types were positive for PAS and toluidine blue, indicating the presence of mucopolysaccharides, whereas semigranular and granular haemocytes were rich in carbohydrates and lipid moieties. Besides, prophenol oxidase was localised within the granules of semigranular and granular haemocytes. Hyaline haemocytes showed an abundance of well differentiated cytoplasmic organelles and granules, and there was a distinct differentiation between semigranular and granular haemocytes in terms of granules and organelles. This is the first report of the characterisation of haemocytes of the mud crab.

Characterization of red sternum syndrome in mud crab farms from Thailand

Biologia ◽  
2010 ◽  
Vol 65 (1) ◽  
Author(s):  
Mayuva Areekijseree ◽  
Thanaporn Chuen-Im ◽  
Busaba Panyarachun
Keyword(s):  
Electron Microscopy ◽  
Cell Walls ◽  
Scylla Serrata ◽  
Mud Crab ◽  
Internal Organs ◽  
Three Stages ◽  
Severity Of The Disease ◽  
Claw Muscle ◽  
Scanning Electron

AbstractSamples of abnormal mud crabs, Scylla serrata (Forskål, 1755) (Decapoda: Portunidae), were collected from crab farms in Samutsongkhram Province, Thailand. These crabs had hard carapaces, red chelipeds and joints, pale hepatopancreas, gills, and soft muscles. They were almost immobile and finally died. The haemolymph revealed three stages of the syndrome, namely orange, orange-white, and milky-white in colors. The haemolymph, integument, hepatopancreas, gills, abdominal and claw muscle, stomach, and heart were dissected and histologically examined using transmission and scanning electron microscopy. Closer examinations found infection with rod-, curve rod-, or coccus-shape bacteria with thin and thick cell walls in all investigated organs and haemolymph. Isolation of the microorganisms from the infected tissues of red sternum syndrome crabs resulted in five types of bacteria. No microorganism growth was observed in normal crabs. Interestingly, the types of isolated bacteria can be classified according to the severity of the disease. Additionally, the degree of bacterial infection found was consistent with the stages of the disease. It was postulated that the bacteria entered the crabs via the gills, and then migrated through circulating haemocytes, before reaching the internal organs.

Light and electron microscopic demonstration by the PATS reaction of peripheral nerve regeneration

1989 ◽  
Vol 47 ◽  
pp. 952-953 ◽  
Author(s):  
B. Giammara ◽  
E. Anderson ◽  
P. Yates ◽  
J. Hanker
Keyword(s):  
Electron Microscopy ◽  
Toluidine Blue ◽  
Periodic Acid ◽  
Light And Electron Microscopy ◽  
Nerve Fibers ◽  
Blue Staining ◽  
Hydroxyl Groups ◽  
Distal Stump ◽  
Thin Sections ◽  
Toluidine Blue Staining

Although periodic acid-Schiff(PAS) type reactions have been applied to nervous tissues for many years, interest has centered upon staining glycolipids, principally myelin constituents such as the class of sphingolipids. The staining of these compounds such as sphingomyelin has generally been attributed to the presence of amino and hydroxyl groups on adjacent carbon atoms of carbohydrate of the sphingosine moiety. But unsaturated lipids also give the reaction and sphingolipids stain even if carbohydrate moieties are absent. This reaction has been used for staining myelin sheaths but lipid solvents must be avoided in processing the specimens. Toluidine blue staining of semi-thin sections of epoxy- embedded nerve specimens has also been widely used to study regenerating fibers after nerve transection or avulsion. A recent study was made in our laboratories of conduits (sleeves) tailored from biodegradable polyester (VicrylR) mesh to guide the reconnection of regenerating fibers from the proximal stump of a rat sciatic nerve, across an 11 mm gap, with fibers in the distal stump of the interrupted nerve. Complete reconnection of the stumps was observed as early as one month after creating the avulsive nerve injury.Comparison of transverse sections of the repaired sciatic with sections of control nerve with the toluidine blue stain, however, showed little evidence of axonal regeneration after one month (Figs. 1,3). A variation of the PAS reaction (depositing silver) for light and electron microscopy developed in our laboratories (PATS reaction, 5) was than applied to the study of the semi-thin sections of the epoxy-embedded control and repaired sciatic nerves of the same rat one month postsurgery. Correlative light and scanning electron microscopy by SEI and BEI modes could then be performed since the PATS reaction produced very satisfactory staining of the semi-thin sections (Figs. 3-5). Myelin was not stained by the PATS reaction in these specimens since the nerves had been processed with lipid solvents for epoxy embedment. Schwann cells, however, were very prominent in control but not in the repaired nerve. The inner layers of endoneurium and all pericapillaries associated with nerve fibers were intensely stained due to their reticulin content in both control and repaired nerve (Figs. 2,4). This was not unexpected because the PATS reaction employs a silver methenamine reagent. Thus, with the PATS reaction axons could be identified in sections of repaired nerve (Fig. 4) that could not be discerned with toluidine blue staining (Fig. 3). In sections of repaired nerve stained with either toluidine blue or the PATS reaction few axons or axis cylinders were observed but more were seen with the PATS stain (Figs. 3,4). In control nerve sections stained with either procedure many were seen (Figs. 1,2).

Aspects ultrastructuraux et cytochimiques des hématies nucléées de deux Annélides Polychètes Notomastus latericeus Sars et Glycera convoluta Keferstein

1980 ◽  
Vol 58 (4) ◽  
pp. 589-597 ◽  
Author(s):  
K. E. Sean ◽  
B. Boilly
Keyword(s):  
Electron Microscopy ◽  
Iron Content ◽  
Light And Electron Microscopy ◽  
Cytoplasmic Organelles ◽  
Glycera Convoluta

The structure and cytochemistry (pseudoperoxidase activity and iron content) of the nucleated erythrocytes of two polychaetes, Notomastus latericeus Sars and Glycera convoluta Keferstein, have been investigated using light and electron microscopy. These cells contain few cytoplasmic organelles but do possess iron-rich structures in vacuoles and free in the cytoplasm. Both species show a pseudoperoxidase activity throughout the cytoplasm and, in the case of Notomastus, in the nucleus as well. The cytological and cytochemical aspects of these cells suggest a low metabolism and a limited lifetime, particularly in Notomastus.

Planothidium galaicum sp. nov. (Bacillariophyta, Achnanthidiaceae), a new diatom species from Galician coast, Spain

Phytotaxa ◽  
2013 ◽  
Vol 151 (1) ◽  
pp. 44 ◽  
Author(s):  
IRENE ÁLVAREZ-BLANCO ◽  
SAÚL BLANCO
Keyword(s):  
Electron Microscopy ◽  
Related Species ◽  
Light And Electron Microscopy ◽  
Morphological Features ◽  
Diatom Species ◽  
Galician Coast ◽  
Himanthalia Elongata ◽  
Frustule Morphology

A new Planothidium species, Planothidium galaicum sp. nov. is described on the basis of light and electron microscopy investigations of its frustule morphology. This diatom was found in the epiphyton of Himanthalia elongata at Muxía coast (Galicia, northwest Spain). A discussion of the morphological features of this taxon and their taxonomic affinities with related species, is presented.

Proteoglycan alterations in rheumatoid arthritis—a light and electron microscopy study

1978 ◽  
Vol 36 (2) ◽  
pp. 676-677
Author(s):  
Nelson S. Mitchell
Keyword(s):  
Rheumatoid Arthritis ◽  
Electron Microscopy ◽  
Toluidine Blue ◽  
Light And Electron Microscopy ◽  
Microscopy Study ◽  
Electron Microscopy Study ◽  
Clear Space ◽  
Staining Techniques ◽  
Femoral Condyles ◽  
Control Samples

We have previously studied the ultramorphology oh cells in rheumatoid arthritis using conventional staining techniques. We have recently reported a technique for the precipitation and fixation of proteoglycan using Toluidine Blue 0 or Safranin 0 which permits simultaneous localization of proteoglycan using either light or electron microscopy in sections cut from the same block. This paper reports a study of rheumatoid arthritic cartilage using this technique.Samples oh cartilage were removed from the femoral condyles of thirteen patients with classical rheumatoid arthritis who were having operations on the knee. Control samples were removed from the knees of eleven normal patients who were having surgery for recent trauma. In both Instances, the tissues were prepared with Toluidine Blue or Safranin 0 Introduced into the fixation process as will be described.When fixed and stained by conventional methods, articular cartilage has been traditionally reported to consist ofi chondrocytes lying in a matrix of collagen and proteoglycan though separated from this matrix by a pericellular clear space, “halo” or “lacuna” of varying dimensions. This pericellular space was thought to contain only small amounts of collagen.

Mouse Hepatoblastomas: A Histologic, Ultrastructural, and Immunohistochemical Study

1988 ◽  
Vol 25 (4) ◽  
pp. 286-296 ◽  
Author(s):  
T. Nonoyama ◽  
F. Fullerton ◽  
G. Reznik ◽  
T. J. Bucci ◽  
J. M. Ward
Keyword(s):  
Electron Microscopy ◽  
Tumor Cells ◽  
Immunohistochemical Study ◽  
Light And Electron Microscopy ◽  
Alpha Fetoprotein ◽  
Cell Populations ◽  
Bile Canaliculi ◽  
Cytoplasmic Organelles ◽  
Dense Bodies ◽  
Hepatocellular Adenomas

Hepatoblastomas from B6C3F1 and BALB/c mice were examined by light and electron microscopy and by immunohistochemical reactions for alpha-fetoprotein, keratin, and vimentin. Tumors occurred in one group of a chronic bioassay for the interaction of diet, genetic strain, and the carcinogen, 2-acetylaminofluorene. Tumors had several populations (including epithelial and mesenchymal cells) in various stages of differentiation. Neoplastic epithelial cells had features of embryonal hepatocytes, such as sparse cytoplasmic organelles, absence of glycogen, abundant free ribosomes, occasional bile canaliculi, and peroxisome-like dense bodies. Embryonal fibroblast-like cells had pleomorphic and folded nuclei with prominent perinuclear chromatin and dispersed cytoplasmic organelles. Fibroblast-like cells were surrounded by bundles of collagen fibrils. Intermediate or transitional types of cells were seen. No tumor cells were immunoreactive for mouse alpha-fetoprotein (AFP) antibody, unlike those in hepatocellular adenomas or carcinomas. Epithelial and mesenchymal tumor cells contained intermediate filaments throughout the cytoplasm; some of these cells stained for keratin but not for vimentin. These findings suggest that mouse hepatoblastomas are derived from bipotential liver blastema cells and are composed of a mixture of several cell populations.

scholarly journals Light and Electron Microscopy of the European Beaver (Castor fiber) Stomach Reveal Unique Morphological Features with Possible General Biological Significance

PLoS ONE ◽  
2014 ◽  
Vol 9 (4) ◽  
pp. e94590 ◽  
Author(s):  
Natalia Ziółkowska ◽  
Bogdan Lewczuk ◽  
Wojciech Petryński ◽  
Katarzyna Palkowska ◽  
Magdalena Prusik ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Biological Significance ◽  
Morphological Features ◽  
Castor Fiber ◽  
European Beaver

scholarly journals Simultaneous localization of proteoglycan by light and electron microscopy using toluidine blue O. A study of epiphyseal cartilage.

1976 ◽  
Vol 24 (5) ◽  
pp. 621-629 ◽  
Author(s):  
N Shepard ◽  
N Mitchell
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Toluidine Blue ◽  
Light And Electron Microscopy ◽  
Uranyl Acetate ◽  
Collagen Fibrils ◽  
Epiphyseal Cartilage ◽  
Fixation Method ◽  
Fixed Tissue

The simultaneous localization of proteoglycan by light and electron microscopy was demonstrated by fixing epiphyseal cartilage in a glutaraldehyde toluidine blue O solution. Sections cut for light microscopy viewing and those cut for electron microscopy required no further staining, although, in the latter case, staining with uranyl acetate and lead improved the overall contrast. By this technique, electron-dense structures were seen concentrated about the cells which were actively synthesizing matrix, and these structures appeared to bind collagen fibrils. Similar structures were not seen in conventionally fixed tissue. They could also not be identified when the specimens were previously incubated with the proteoglycan-digesting enzyme, papain, prior to toluidine blue O fixation. The toluidine blue O fixation method, unlike conventional fixation and staining, retained proteoglycan in the pericellular areas of actively synthesizing cells and made it visible by light and electron microscopy. It appears that proteoglycans is both precipitated and stained by the presence of toluidine blue O during fixation.

A Simple Dichromatic Stain for Plastic Embedded Tissues

1970 ◽  
Vol 28 ◽  
pp. 296-297 ◽  
Author(s):  
G. R. Mackay ◽  
M. L. Mead
Keyword(s):  
Electron Microscopy ◽  
Toluidine Blue ◽  
Biological Material ◽  
Good Reproducibility ◽  
Routine Work ◽  
Staining Techniques

Color contrasting of 1 to 2 micron sections of plastic embedded biological material is an important adjunct to electron microscopy. The procedures in general use today are simple and rapid giving monochromatic results, e.g., toluidine blue. Although many di- and polychromatic histologic staining techniques have been modified to obtain a counterstaining effect with plasticembedded tissue, the methods are usually undesirable for routine work because they are time consuming, complicated and often defy good reproducibility.

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