A comparison of methodologies for the staining and quantification of intracellular components of Arbuscular Mychorrizal (AM) fungi in the root cortex of two varieties of winter wheat

© 2019 The Authors. The definitive peer reviewed, edited version of this article is published in Access Microbiology, https://doi.org/10.1099/acmi.0.000083. This is an open-access article distributed under the terms of the Creative Commons Attribution License.Arbuscular Mychorrizal (AM) fungi are one of the most common fungal organisms to exist in symbiosis with terrestrial plants facilitating the growth and maintenance of arable crops. Wheat has been studied extensively for AM fungal symbiosis using the carcinogen trypan blue as the identifying stain for fungal components, namely arbuscles, vesicles and hyphal structures. The present study uses Sheaffer® blue ink with a lower risk as an alternative to this carcinogenic stain. Justification for this is determined by stained wheat root sections (n = 120), with statistically significant increases in the observed abundance of intracellular root cortical fungal structures stained with Sheaffer® blue ink compared to trypan blue for both Zulu (P = 0.003) and Siskin (P = 0.0003) varieties of winter wheat. This new alternative combines an improved quantification of intracellular fungal components with a lower hazard risk at a lower cost.Peer reviewe

Influence of blue stain on density and dimensional stability of Pinus radiata timber from northern Galicia (Spain)

Holzforschung. International Journal of the Biology, Chemistry, Physics, and Technology of Wood; 69(1): 97–102 DOI: 10.1515/hf-2014-0014The presence of blue-stain fungi considerably decreases the value of Pinus radiata timber, which is commercially important in Galicia (NW Spain). For this study, seven young pine trees growing in four different plantations were felled, and 20 discs were sampled from different heights in the stems. Parts of the discs were discoloured as a result of fungal infection. The discs were cut into small specimens. Defect-free specimens (controls) were selected for determining density and dimensional stability both in the volume and in the axial direction. Physical properties of fully blue-stained specimens and the controls were compared for each disc based on one-factor analysis of variance. As the factor was blue-stain, the other sources of variation were suppressed, such as tree source, height in the stem, ring width, cambial age and presence of heartwood or sapwood. Most of the physical properties analysed on some sample discs differed significantly between discoloured and unstained wood. The variables most affected by blue stain were basic density and volumetric shrinkage: blue-stained wood was 1.1% lighter and volumetric shrinkage was 5% higher in blue-stained wood than in the controls

Organic contamination of LDEF

A brown stain of varying thickness was present on most of the exterior surface of the retrieved Long Duration Exposure Facility (LDEF). Tape lifts of Earth-end LDEF surfaces taken in Feb. 1990 showed that the surface particle cleanliness immediately after retrieval was very good, but faint footprints of the tape strips on the tested surfaces indicated a very faint film was removed by the tape. Solvent wipes of these surfaces showed that the stain was not amenable to standard organic solvent removal. Infrared spectra of optical windows from tray E5 and scrapings indicate that the film is primarily of organic composition, but is not similar to the oil that seeped from tray C12. Very dark and heavy deposits of the stain are present at openings and vents to the interior of the LDEF. Heavy brown and blue-green deposits are present in the interior of LDEF where sunlight penetrated through cracks and vent openings. Photographs of the deintegrated LDEF graphically show the stain distribution. The exterior of the LDEF had significant areas painted with a white polyurethane paint for thermal control, and almost all of the interior was painted with a black polyurethane paint for thermal control. The brown staining of the LDEF is consistent with long-term outgassing of hydrocarbons from these paints followed by rapid solar-ultraviolet-induced polymerization of the outgassed hydrocarbons when the outgassed molecules stuck to surfaces exposed to sunlight

Lipid, detergent, and coomassie blue G-250 affect the migration of small membrane proteins in blue native gels:Mitochondrial carriers migrate as monomers not dimers

Background: Mitochondrial carriers were thought to be dimeric based on their migration in blue native gels.  Results: The high molecular mass species observed in blue native gels are composed of protein monomers, detergent, lipid, and Coomassie stain.  Conclusion: The mitochondrial carriers are monomeric not dimeric.  Significance: The apparent mass of small membrane proteins in blue native gels requires significant correction

New Tetrachromic VOF Stain (Type III-G.S) for Normal and Pathological Fish Tissues

10 páginas, 3 figuras, 1 tabla. Este trabajo esta dedicado a Sra. María Armenta.A new VOF Type III-G.S stain was applied to histological sections of different organs and tissues of healthy and pathological larvae, juvenile and adult fish species (Solea senegalensis; Sparus aurata; Diplodus sargo; Pagrus auriga; Argyrosomus regius and Halobatrachus didactylus). In comparison to the original Gutiérrez VOF stain, more acid dyes of contrasting colours and polychromatic/metachromatic properties were incorporated as essential constituents of the tetrachromic VOF stain. This facilitates the selective staining of different basic tissues and improves the morphological analysis of histochemical approaches of the cell components. The VOF Type III -6.5 stain is composed of a mixture of several dyes of varying size and molecular weight (Orange G<acid Fuchsin<Light green<Methyl Blue<Fast Green), which are used simultaneously, and it enables the individual tissues to be selectively differentiated and stained. Muscle fibers, collagen, reticulin and elastin fibers, erythrocytes, cartilage, bone, mucous cells, oocytes and larvae were selectively stained and differentiated. Dyes with small size and molecular weight (i.e Orange G), penetrate all tissue structures rapidly, but are only tightly retained in densely textured tissues (i.e erythrocytes). Methyl Blue is an interesting triarylmethane dye (large size and molecular weight), which is incorporated in this new VOF tetrachrome stain, and acquires histochemical significance when used at acid pH (2.8) because collagen and reticulin fibers, as well basophilic and metachromatic substances (strongly ionized sulphated glycoconjugates) can be identified. Muscle tissues show an evident green colour (Fast Green or Light Green affinities), even those isolated and/or diffuse muscle fibers present in the digestive submucosa layer. Connective tissues showed a specific and strong blue colour (Methyl Blue affinity) or mixed blue-red staining (Methyl Blue and Acid Fucshin affinities). Very noticeable is the staining of the mucous cells, as well as the hyaline capsule of the viral lymphocystic cells, which were stained blue-purple (carboxylated and/or strongly ionized sulphated groups). Cartilaginous tissues showed a blue or purple (Methyl Blue affinity) staining, and a specific red colour (Acid Fucshin affinity) was evident during calcification or in bone structures (i.e skeleton, fins, gills, teeth).This work (Spanish MCYT/AGL2003-03558).Peer reviewe

Comparision of staining methods for two dimensional electrophoresis gel resolved with Puntius javanicus liver proteome

The aim of this study was to compare the various staining methods based on commassie briliant blue and silver nitrate stain for the two dimensional gel electrophoresis resolved with Puntius javanicus liver proteome. The staining methods were selected base on the previous reportabout their compatiblitywiththe mass spectrometry analysis. Sliver staining methodis known as the most sensitive method to visualize the maximum number of protein spots resolved in 2D gel but it is less sensitive(incompatible) toward mass spectrometry detection. Results of this study showed that a modifiedstaining method using colloidal coomassie blue G-250 (CCB) is roughly similarly sensitive but lower protein spot detected compared with silver staining (SS) as indicated at the number of 303±26 and 693±14of proteinresolved in both types of stained gels. The conventional methods of staining using commassie brilliant blue G-250 and R-250 only detected less number of protein spots(128±17and 78±11, respectively) compared to modified CCB staining method. As the commasie brilliant blue stain is known to be a very sensitive for mass spectrometry detection, the modified method of CCB was selected for further study on Puntius javanicus liver proteome

Chemical enhancement of footwear impressions in blood recovered from cotton using alginate casts

Depletion series of footwear impressions in blood were deposited on black cotton fabric after which they were lifted using alginate and subsequently enhanced using protein stains amido black (AB), Crowle’s stain (CS), coomassie blue (CB), and Hungarian red (HR). Other factors that were considered during this study were the age of the impression and the temperature of the environment. A novel score system for the enhancement of footwear impressions was introduced, which used the product of scores for size and detail of the impression. The study showed that temperatures between 8 °C and 37 °C did not impact chemical enhancement, whereas the age of the impression did. An impression aged for 7 days yielded higher enhancement scores than impressions aged for 1 or 28 days, especially for AB. The results of depletions 1 to 5 were similar to the results of only depletion 5. However, at depletion 5, AB was the best-performing protein stain. CB and AB yielded the highest level of enhancement of the impressions, whereas CS and HR resulted in poorer quality enhancements. AB was the preferred protein stain of use because AB was the most sensitive protein stain used in this study and there were fewer health risks involved in using water-based AB than in using methanol-based CB

Influence of biotic factors on the mechanical properties of wood, taking into account the time of harvesting

The aim of the present paper is to investigate the influence of biotic factors (fungi and insects) on the mechanical properties of wood through the effect of blue-stain, taking into account the time of harvesting and the time of stay of wood in the forest. Specifically, the resistance to axial compression and to bending (modulus of rupture (MOR)) was studied using infected specimens of Scots Pine (Pinus sylvestris) and Norway Spruce (Picea abies) (the usual types of wood used in woodwork). The specimens were obtained from logs of Scots Pine and Norway Spruce that were harvested in three different seasons of year, namely in July 2012, November 2012 and June 2013, respectively, in the forest of Elatia-Greece, and the attack pace by biotic factors with respect to the time of logging was studied. The placement of the experimental surfaces of each type of tree was made on skid road and in the stand. Totally, 120 laboratory measurements in axial compression and 120 measurements in bending (MOR) took place. The results proved that blue-stain hardly affect the mechanical properties of both wooden species and particularly the specimens that were derived during the winter logging

A Monolithically Fabricated Combinatorial Mixer for Microchip-Based High-Throughput Cell Culturing Assays

We present an integrated method to fabricate 3- D microfluidic networks and fabricated the first on-chip cell culture device with an integrated combinatorial mixer. The combinatorial mixer is designed for screening the combinatorial effects of different compounds on cells. The monolithic fabrication method with parylene C as the basic structural material allows us to avoid wafer bonding and achieves precise alignment between microfluidic channels. As a proof-of-concept, we fabricated a device with a three-input combinatorial mixer and demonstrated that the mixer can produce all the possible combinations. Also, we demonstrated the ability to culture cells on-chip and performed a simple cell assay on-chip using trypan blue to stain dead cells