Evaluation of a vital staining protocol with 2,3,5-triphenyltetrazolium chloride for cancellous bone in a sheep model

Turnip (Brassica rapa L.) root tissue was exposed to freeze-thaw stresses of −7, −9, -−11 and −19 °C. The post-thawed tissues were either subjected to leaching in deionized water for 2 h or left at 100% humidity. Tissue survival was then assayed by vital staining using modified 2,3,5-triphenyltetrazolium chloride (TTC) staining in 0.2 M phosphate buffer. Tissue survival was significantly lower for leached samples than for non-leached samples. It is concluded that freeze-thaw injury in plant tissues is enhanced by post-thaw leaching in water. The 0.05 M phosphate buffer commonly used for TTC staining also damaged freeze-thaw injured tissue. Key words:Brassica rapa L., 2,3,5-triphenyltetrazolium chloride (TTC), freeze-thaw injury, leaching

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A Sheep Model for Cancellous Bone Healing

Frontiers in Surgery ◽

10.3389/fsurg.2014.00037 ◽

2014 ◽

Vol 1 ◽

Cited By ~ 14

Author(s):

Angad Malhotra ◽

Matthew Henry Pelletier ◽

Yan Yu ◽

Chris Christou ◽

William Robert Walsh

Keyword(s):

Cancellous Bone ◽

Bone Healing ◽

Sheep Model

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Asymbiotic in vitro germination and seed quality assessment of Australian terrestrial orchids

Australian Journal of Botany ◽

10.1071/bt12133 ◽

2012 ◽

Vol 60 (7) ◽

pp. 592 ◽

Cited By ~ 11

Author(s):

Nicole Dowling ◽

Manfred Jusaitis

Keyword(s):

Seed Quality ◽

Culture Media ◽

Vital Staining ◽

Terrestrial Orchid ◽

Triphenyltetrazolium Chloride ◽

Asymbiotic Seed Germination ◽

Orchid Species ◽

Terrestrial Orchids ◽

Asymbiotic Germination

Determining the seed quality and germination requirements for threatened orchid species in storage is vital for future conservation efforts. Seeds of many Australian terrestrial orchid species are held in conservation collections around the country, but few have been germinated in vitro, fuelling concerns over their long-term viability. This study tested three methods of assessing orchid seed quality; asymbiotic germination was compared with vital staining using triphenyltetrazolium chloride or fluorescein diacetate. Six culture media were examined for efficacy in promoting asymbiotic seed germination of four Australian terrestrial orchid species (Pterostylis nutans, Microtis arenaria, Thelymitra pauciflora and Prasophyllum pruinosum). Germination occurred on all media but germination rates were consistently highest on BM1 and development was most advanced on BM1, P723 and Malmgren media. Subsequent trials tested the efficacy of BM1 for asymbiotic germination of additional genera (Caladenia, Calochilus and Diuris), several congeneric species, and two species collected from several different provenances within each of their ranges. The results indicate that asymbiotic germination on BM1 medium is an effective technique for testing the performance of Australian terrestrial orchid seeds. The efficacy of vital stains to determine seed viability, however, remains uncertain, as significant disagreement between degree of staining and germinability was observed for some species.

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OBSERVATIONS ON THE USE OF TETRAZOLIUM SALTS IN THE VITAL STAINING OF BACTERIA

Canadian Journal of Microbiology ◽

10.1139/m59-028 ◽

1959 ◽

Vol 5 (3) ◽

pp. 245-250 ◽

Cited By ~ 10

Author(s):

L. Eidus ◽

B. B. Diena ◽

L. Greenberg

Keyword(s):

Bacterial Growth ◽

Vital Staining ◽

Nitro Blue Tetrazolium ◽

Complete Inhibition ◽

Triphenyltetrazolium Chloride ◽

Tetrazolium Salts ◽

Blue Tetrazolium

Studies on the vital staining of bacteria were carried out to determine the concentrations of tetrazolium salts required for optimal staining, and the amounts which could cause complete inhibition of bacterial growth. Four salts were studied: neotetrazolium chloride (NTC), triphenyltetrazolium chloride (TTC), blue tetrazolium (BT), and nitro-blue tetrazolium (NBT). Evidence has been presented to show that only living, actively metabolizing cells can reduce neotetrazolium to formazan.

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424 A Comparison of Techniques Used for Determining Freezing Injury on Two Apple (Malus × domestica Borkh.) Cultivars: Liberty and RedMax

HortScience ◽

10.21273/hortsci.35.3.466c ◽

2000 ◽

Vol 35 (3) ◽

pp. 466C-466

Author(s):

M.E. Garcia ◽

Linda A. Boccuzzo

Keyword(s):

Block Design ◽

Vital Staining ◽

Fruit Trees ◽

Early Spring ◽

Freezing Injury ◽

Testing Time ◽

Triphenyltetrazolium Chloride ◽

Deciduous Fruit ◽

Comparison Of Techniques ◽

Artificial Freezing

Hardiness testing of the wood of deciduous fruit trees has been conducted using a variety of techniques. In our studies, the objective was to determine an efficient method of determining freezing injury for apple (Malus × domestica Borkh.) wood. We tested 1-year old wood of two cultivars: Liberty and RedMax. The wood was tested over the course of 2 years (1998 and 1999). Collection began in the late fall and continued throughout the winter (until it was determined full hardiness had been achieved) and then again in the early spring. The wood was cut into 1-cm sections and frozen. The artificial freezing was conducted in an ethanol bath, with the temperature lowered at 5 °C/h. Samples were removed in 3-min intervals. After freezing, the wood was acclimated to 4 °C for 12 h. Three tests were conducted to determine the hardiness/injury to the tissues. The tests used were: discoloration, callus growth and vital staining (with 2,3,5-triphenyltetrazolium chloride). This was a split block design with samples collected randomly from each tree. Four replicates (12 trees) of each cultivar were tested. Results showed that the callus test predicted the same LT50 as the other two tests, discoloration and vital staining. Discoloration was not easy to differentiate and was the most time-consuming. The callus grown by the apple wood was easily formed and distinguished. The callus test does not require the tetrazolium stain; therefore, one less step was needed in comparison to the vital staining test. This reduced testing time by over 6 h.

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553 PB342 COLD RESISTANCE OF `ANJOU', `BARTLETT' AND `BOSC' PEAR TREES

HortScience ◽

10.21273/hortsci.29.5.510f ◽

1994 ◽

Vol 29 (5) ◽

pp. 510f-510

Author(s):

Delmer O. Ketchie ◽

R. Kammereck

Keyword(s):

Cold Resistance ◽

Vital Staining ◽

Limited Growth ◽

Triphenyltetrazolium Chloride ◽

Current Season ◽

Freeze Resistance ◽

Additional Resistance ◽

Pear Trees ◽

Sampling Times ◽

Better Than

Samples of current season shoots of Anjou, Bartlett and Bosc pears were collected throughout the year during 1990, `91 and `92. Differential thermal analysis (DTA) and vital staining with triphenyltetrazolium chloride (TTC) were used at the sampling times to determine freeze resistance. Freezing tests were conducted on greenhouse-grown trees. Temperatures to freeze the trees were predetermined by DTA. After freezing TTC staining, acid fuchsin test and growth were used to determine survival. All three varieties began to acclimate after terminal growth ceased in late June until October. Bartlett and Anjou obtained about -25°C resistance by this time and Bose about -23°C. After frost began, Anjou and Bartlett gained an additional resistance to -33°C and Bose to -28°C. Trees frozen artificially at -27°C had limited growth but did leaf out only to die a month later. Trees frozen at -33°C never leafed out Bartlett trees at -27°C looked better than Anjou and Bose trees but died also.

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Fading of the diaminobenzidine reaction product in the confocal scanning laser microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100152707 ◽

1989 ◽

Vol 47 ◽

pp. 146-147

Author(s):

JS Deitch ◽

KL Smith ◽

JW Swann ◽

JN Turner

Keyword(s):

Pyramidal Neurons ◽

Light And Electron Microscopy ◽

Scanning Laser ◽

Laser Exposure ◽

Confocal Scanning Laser Microscope ◽

Fluorescent Markers ◽

Before And After ◽

Staining Protocol ◽

Confocal Scanning ◽

Confocal Scanning Laser

Neurons labeled with horseradish peroxidase and reacted with diaminobenzidine (DAB) can be imaged using a confocal scanning laser microscope (CSLM) in the reflection mode. In contrast to fluorescent markers, the DAB reaction product is thought to be stable and can be observed by both light and electron microscopy. We have investigated the sensitivity of the DAB reaction product to laser irradiation, and present the spectrophotometric properties of DAB before and after exposure in the CSLM.Pyramidal neurons in slices of rat hippocampus were injected with biocytin (a biotin-lysine complex), fixed overnight in 4% paraformaldehyde, and vibratome sectioned at 75 μm. Biocytin was detected with avidin-HRP (1:200) in 0.5% Triton X-100, incubated in DAB (0.5 mg/ml) with or without 0.04% nickel ammonium sulfate (Ni), dehydrated, and imaged in a Bio Rad MRC-500 CSLM with an argon ion laser (488 and 514 nm). Spectrophotometric measurements of the soma were made on a Zeiss microspectrophotometer, as a function of laser exposure (100-1000 scans) and staining protocol.

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