A Quick and Simple Method to Evaluate Anisogramma anomala Ascospore Viability

Viability of ascospores of Anisogramma anomala, cause of eastern filbert blight on European hazelnut, was assessed using the vital stain trypan blue (working solution of 0.05% in lactoglycerol). Viable ascospores only had faint blue staining around their cell walls while non-viable ascospores absorbed the stain and turned dark blue. The number of viable (non-stained) ascospores as determined by trypan blue was similar to the proportion of ascospores germinating on culture media. Viability of field collected ascospores from rainwater spore traps ranged from 41 to 68%. Disease incidence of hazelnut seedlings was more closely related to differences in ascospore abundance than to differences in ascospore viability. Accepted for publication 3 January 2013. Published 9 May 2013.

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Disease Incidence and Ascospore Dispersal from Cut Hazelnut Branches Colonized by Anisogramma anomala

Plant Disease ◽

10.1094/pdis-06-13-0631-re ◽

2014 ◽

Vol 98 (6) ◽

pp. 834-838 ◽

Cited By ~ 4

Author(s):

S. Heckert ◽

J. W. Pscheidt ◽

S. A. Cluskey

Keyword(s):

Trypan Blue ◽

Disease Incidence ◽

Bud Break ◽

Splash Dispersal ◽

Anisogramma Anomala ◽

Blue Stain ◽

Rain Splash

Hazelnut branches bearing stromata of Anisogramma anomala cut in December (2009 and 2010) were compared with branches cut prior to bud break in March to investigate these sources of inoculum. Branches were placed into brush piles (sources). Spore traps and potted hazelnut trees were placed adjacent to each source, 6.4 m upwind and downwind, and 20 m downwind from each source. Significantly more ascospores were detected near sources of branches cut in March compared with December in 2010 however, no differences were detected between pruning treatments in 2011. Ascospore viability, as assessed by trypan blue stain, averaged 50% for both pruning times each season. Significantly more ascospores were detected 6.4 m downwind compared with 6.4 m upwind or 20 m downwind of a source both years. All potted trees exposed to branches from both pruning treatments within sources became diseased both years. The proportion of potted trees that became infected was greater for the downwind group than the upwind for both years, suggesting that ascospores were dispersed beyond the rain splash dispersal range of sources. Ascospores from diseased branches pruned in December or March remained viable, infectious and were dispersed at least 20 m downwind.

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Use of a trypan blue assay to measure the deoxyribonucleic acid content and radioactive labeling of viable cells.

Journal of Histochemistry & Cytochemistry ◽

10.1177/28.7.6156203 ◽

1980 ◽

Vol 28 (7) ◽

pp. 700-703 ◽

Cited By ~ 28

Author(s):

D C Allison ◽

P Ridolpho

Keyword(s):

Trypan Blue ◽

Isotopic Labeling ◽

Blue Color ◽

Simple Method ◽

Viable Cells ◽

Cellular Incorporation ◽

Dark Blue ◽

Thymidine Label ◽

Selection Of

A simple method of determining cellular viability, DNA content, and isotopic labeling is described. Cells are sedimented onto slides and fixed in a mixture of trypan blue and paraformaldehyde which stains nonviable cells a dark blue color, whereas viable cells exclude the dye. The staining pattern is stable for at least 6 months and persists after the Feulgen reaction, allowing the selection of viable cells for cytophotometric DNA determinations. We ascertained cellular incorporation of a 3H-thymidine label by photomapping cells prior to cytophotometry and relocating these cells after preparation of autoradiographs. The method gives accurate quantitative DNA and labeling values of mouse thymocytes labeled in vitro and may be of value in studies of tumors containing a large proportion of necrotic cells.

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Trypan blue staining in vitreoretinal surgery

Ophthalmology ◽

10.1016/j.ophtha.2004.05.008 ◽

2004 ◽

Vol 111 (8) ◽

pp. 1622-1623 ◽

Cited By ~ 3

Author(s):

Gema Rebolleda ◽

Francisco José Muñoz Negrete ◽

Marta Suarez-Figueroa

Keyword(s):

Trypan Blue ◽

Vitreoretinal Surgery ◽

Blue Staining ◽

Trypan Blue Staining

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Use of trypan blue staining to assess the quality of ovarian cryopreservation

Fertility and Sterility ◽

10.1016/j.fertnstert.2006.08.115 ◽

2007 ◽

Vol 87 (5) ◽

pp. 1200-1207 ◽

Cited By ~ 50

Author(s):

Patricia Fauque ◽

Anis Ben Amor ◽

Christiane Joanne ◽

Germain Agnani ◽

Jean Luc Bresson ◽

...

Keyword(s):

Trypan Blue ◽

Blue Staining ◽

Trypan Blue Staining

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An Examination of Baker's Acid Haematein Test for Phospholipines

Journal of Cell Science ◽

10.1242/jcs.s3-88.4.467 ◽

1947 ◽

Vol s3-88 (4) ◽

pp. 467-478

Author(s):

A. J. CAIN

Keyword(s):

Calcium Ions ◽

The Other ◽

Blue Staining ◽

Other Substances ◽

Whole Class ◽

Dark Blue ◽

General Explanation

1. Baker's acid haematein test for phospholipines is specific provided that only a definite positively result is considered. Very pale blues and greys may be caused by other lipoids, which if present in very large masses may possibly show medium to dark blue granules but will not be coloured all through. 2. The mechanism of the test appears to be as follows: (a) Phospholipine is not fixed by formal-calcium but is restrained from passing into solution by the calcium ions, which play no other part. (b) Phospholipine combines readily with chromium from the mordanting fluid, and is then rendered insoluble and mordanted. Other substances, acidic and usually containing phosphorus, are mordanted as well. (c) On staining, blue and brown colorations are formed; in both cases the dye attaches itself to the chromium in the various substrates. (d) On differentiation, some browns and most blues, particularly those with phosphoric substrates, remain nearly fast, but most browns and the weak blues of certain lipoids (not phospholipines) are greatly reduced or removed entirely. The period of differentiation must not be shortened. (e) Blue-staining lipoids (phospholipines) are distinguished from other blue-staining substances by an extraction with the lipoid solvent pyridine, after special fixation. The other substances, and any bound lipoid not removable with pyridine, remain. 3. Since the specificity of the test depends on the relatively greater affinity of phospholipines among lipoids for the mordant, the period of chroming must not be lengthened. 4. One reason why some substances are coloured after pyridine extraction but not after acid haematein is that in the former case they are precipitated and so concentrated; in the latter they are not. This is not a general explanation for the whole class of such substances.

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Inadvertent Trypan Blue Staining of Posterior Capsule during Cataract Surgery Associated with “Argentinian Flag” Event

Case Reports in Ophthalmological Medicine ◽

10.1155/2016/9025063 ◽

2016 ◽

Vol 2016 ◽

pp. 1-3

Author(s):

Robert A. Prinzi ◽

Neeti M. Alapati ◽

Shawn S. Gappy ◽

Jason S. Dilly

Keyword(s):

Cataract Surgery ◽

Trypan Blue ◽

Rare Complication ◽

Posterior Capsule ◽

Blue Staining ◽

Anterior Surface ◽

Anterior Capsule ◽

Trypan Blue Staining ◽

First Case

Trypan blue is common in visualizing the anterior capsule during cataract surgery. Inadvertent staining of the posterior capsule during phacoemulsification is a rare complication and there are few reports in the literature. The proposed mechanism of posterior capsule staining in previous reports includes a compromised zonular apparatus or iris retractors facilitating the posterior flow of trypan blue. We report the first case of trypan blue staining of the posterior capsule associated with the “Argentinian flag” sign. In our case, the “Argentinian flag” allowed the trypan blue to seep between the posterior capsule and the lens, staining the anterior surface of the posterior capsule.

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A simple method of suspending implant wear particles for more physiological modelling of cell-particle interactions in vitro

10.1101/2021.01.05.425343 ◽

2021 ◽

Author(s):

Christine Poon

Keyword(s):

Wear Debris ◽

Culture Media ◽

Polymeric Materials ◽

Wear Particle ◽

Physicochemical Characteristics ◽

Wear Particles ◽

Simple Method ◽

Implant Wear

AbstractArthroplasty implants e.g. hip, knee, spinal disc sustain relatively high compressive loading and friction wear, which lead to the formation of wear particles or debris between articulating surfaces. Despite advances in orthopaedic materials and surface treatments, the production of wear debris from any part of a joint arthroplasty implant is currently unavoidable. Implant wear debris induces host immune responses and inflammation, which causes patient pain and ultimately implant failure through progressive inflammation-mediated osteolysis and implant loosening, where the severity and rate of periprosthetic osteolysis depends on the material and physicochemical characteristics of the wear particles. Evaluating the cytotoxicity of implant wear particles is important for regulatory approved clinical application of arthroplasty implants, as is the study of cell-particle response pathways. However, the wear particles of polymeric materials commonly used for arthroplasty implants tend to float when placed in culture media, which limits their contact with cell cultures. This study reports a simple means of suspending wear particles in liquid medium using sodium carboxymethyl cellulose (NaCMC) to provide a more realistic proxy of the interaction between cells and tissues to wear particles in vivo, which are free-floating in synovial fluid within the joint cavity. Low concentrations of NaCMC dissolved in culture medium were found to be effective for suspending polymeric wear particles. Such suspensions may be used as more physiologically-relevant means for testing cellular responses to implant wear debris, as well as studying the combinative effects of shear and wear particle abrasion on cells in a dynamic culture environments such as perfused tissue-on-chip devices.

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Η χρήση του πεπτιδίου Ec της ισόμορφης IGF-1Ec στην επισκευή χόνδρινων βλαβών

10.12681/eadd/43322 ◽

2018 ◽

Author(s):

Νικόλαος Αρμακόλας

Keyword(s):

Polymerase Chain Reaction ◽

Trypan Blue ◽

Quantitative Polymerase Chain Reaction ◽

Blue Staining ◽

Alcian Blue Staining ◽

Chain Reaction ◽

Polymerase Chain ◽

Tgf Β1 ◽

Invasion Assays

Το πεπτίδιο Ec (PEc) του IGF-1Ec (IGF-1Ec) επάγει την κινητοποίηση των ανθρωπίνων μεσεγχυματικών βλαστικών κυττάρων (hMSC) και ενεργοποιεί την εξωκυτταρική κινάση 1 και 2 (ERK 1/2) διαφόρων κυττάρων. Σκοπός της παρούσας μελέτης ήταν η διερεύνηση της επιδρασης του PEc στην κινητοποίηση και τη διαφοροποίηση των hMSCs, καθώς και η δυνατότητα εφαρμογής του σε συνδυασμό με τον TGF-β1 (TGF-β1) στην επιδιόρθωση του αρθρικού χόνδρου. Τα αποτελέσματα της εξωγενούς χορήγησης του ΡΕc και του ΤGF-β1, ξεχωριστά και σε συνδυασμό, σε hMSCs εκτιμήθηκαν χρησιμοποιώντας trypan blue assay, reverse transcription-quantitative polymerase chain reaction, western blot analysis, Alcian blue staining, wound healing assays και migration/invasion assays. Προσδιορίστηκε ότι το PEc εμπλέκεται στη διαδικασία διαφοροποίησης των hMSCs προς υαλώδη χόνδρο. Η χορήγηση PEc ή / και TGF-β1 σε hMSCs έδειξε συγκρίσιμη εναπόθεση χονδρικής θεμέλειας ουσίας. Ακόμα, η χορήγηση του ΡΕc σε συνδυασμό με τον ΤGF-β1 συσχετίστηκε με μια σημαντική αύξηση στην κινητοποίηση των hMSC σε σύγκριση με την χορήγηση μόνο του TGF-β1 ή του ΡEc (Ρ <0,05). Επομένως, το ΡΕc φαίνεται να διευκολύνει in vitro την κινητοποίηση των hMSC και την διαφοροποίηση τους προς χονδροκύτταρα, ενισχύοντας το ρόλο του ΤGF-β1.

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Quantification of polysaccharides fixed to Gram stained slides using lactophenol cotton blue and digital image processing

F1000Research ◽

10.12688/f1000research.5779.2 ◽

2015 ◽

Vol 4 ◽

pp. 1 ◽

Cited By ~ 5

Author(s):

Bryan Ericksen

Keyword(s):

Image Processing ◽

Digital Image Processing ◽

Digital Image ◽

Blue Staining ◽

Bacterial Cells ◽

Cotton Blue ◽

Specific Polysaccharide ◽

The Media ◽

Light Staining ◽

Dark Blue

Indigo rings and circles emerged when I added the non-specific polysaccharide stain lactophenol cotton blue to Gram stained slides. I attribute the dark blue staining to the presence of capsular polysaccharides and bacterial slime associated with clumps of Gram-negative bacteria. Since all bacterial cells are glycosylated and concentrate polysaccharides from the media, the majority of cells stain light blue. The contrast between dark and light staining is sufficient to enable a digital image processing thresholding technique to be quantitative with little background noise. Prior to the addition of lactophenol cotton blue, the Gram-stained slides appeared unremarkable, lacking ubiquitous clumps or stained polysaccharides. Adding lactophenol cotton blue to Gram stained slides is a quick and inexpensive way to screen cell cultures for bacterial slime, clumps and biofilms that are invisible using the Gram stain alone.

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