A Note on the Staining of the Skeleton of Cleared Specimens with Alizarin Red S

Dawson, Alden B.

doi:10.3109/10520292609115636

Cited by 964 publications

(224 citation statements)

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“…Schultze (1897), probably the first to prepare transparent specimens, used potassium hydroxide, Spalteholz (1911) used none, whereas St Amand & St Amand (1950) used boiling water. Potassium hydroxide has been used in strengths from 1 per cent (Dawson, 1926) to 10 per cent (Jensh & Brent, 1966). It was considered that a more powerful macerating agent might accelerate processing, and the experimental work has confirmed this.…”

Section: Ims Rinsementioning

confidence: 62%

“…A major part of such work involves investigation of the effects of drugs on osteogenesis, for which some modification of Dawson's technique (Dawson, 1926) is commonly used. Essentially the method depends upon maceration of the foetus with alkali, followed by staining with the anthraquinone dye alizarin red S, which has a strong affinity for calcium salts.…”

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Automation of skeletal staining in the evaluation of teratogenicity

Randle¹

1969

Lab Anim

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SUMMARYA number of variables affecting the quality of foetal mouse preparations stained with alizarin red S have been investigated.A seven-day 'Histokinette' procedure has been developed to macerate and stain foetal mice by a modification of Dawson's method.The thalidomide tragedy focussed attention upon the potential teratogenic activity of drugs and this has led not only to a widespread interest in the techniques available but also to a progressive increase in the number of foetuses examined.A major part of such work involves investigation of the effects of drugs on osteogenesis, for which some modification of Dawson's technique (Dawson, 1926) is commonly used. Essentially the method depends upon maceration of the foetus with alkali, followed by staining with the anthraquinone dye alizarin red S, which has a strong affinity for calcium salts. It is capable of producing specimens in which the vividly stained red bone is easily studied through the transparent colourless integument. Experience, however, is necessary to determine the optimal duration of each stage, particularly that of maceration.Inadequate alkali treatment results in a poorly differentiated semi-opaque specimen, whereas excessive exposure may lead to a well-stained foetus that disintegrates on examination.The regular appraisal and repeated handling of each foetus during processing makes the routine use of the method both tedious and time-consuming.In view of this, the possibility of automating the procedure was considered. Such a process which requires a sequence of steps of fixed duration must also provide a satisfactory specimen from both immature and mature foetuses, since both occur in teratological experiments.The investigations reported here were designed to establish the effects of variations in processing technique and in foetal maturity on the quality of the stained specimens.

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Section: Ims Rinsementioning

confidence: 62%

Section: Discussionmentioning

confidence: 99%

“…

…”

mentioning

confidence: 99%

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Automation of skeletal staining in the evaluation of teratogenicity

Randle¹

1969

Lab Anim

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“…A detailed external examination of each viable fetus was conducted. For each litter, half the fetuses were cleared with dilute potassium hydroxide solution, stained with Alizarin Red S, and examined for abnormalities in skeletal development (Dawson, 1926;Staples and Schnell, 1964). The remaining fetuses were fixed in Bouin's solution.…”

Section: Observations and Measurementsmentioning

confidence: 99%

Reproductive consequences of oral arsenate exposure during pregnancy in a mouse model

Hill

Wlodarczyk

Finnell

2008

Birth Defects Research Pt B

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BACKGROUND:The second most common of all structural birth defects, neural tube defects (NTDs), affect approximately 2.6/1,000 births worldwide, and 1/1,000 births in the United States. Of the many environmental agents suspected of being teratogenic, arsenic (As) is capable of inducing NTDs in laboratory animals. METHODS: We evaluated the teratogenicity of oral exposure on embryonic day (E) 7.5 and E:8.5 to As 4.8, 9.6, or 14.4 mg/kg (as sodium arsenate) in an inbred mouse strain, LM/Bc/Fnn, that does not exhibit spontaneous neural tube malformations. Control and arsenic-treated dams (20 per treatment group) were weighed daily, and evaluated for signs of maternal toxicity. Fetuses were evaluated for soft tissue and skeletal malformations. RESULTS: There was no maternal toxicity as evidenced by losses in maternal body weight following As treatment. However, liver weights were lower in all As-treated groups, suggesting hepatotoxicity due to As exposure. The number of litters affected with an NTD (exencephaly) in each treatment group was: 0, 1, 5, and 9 for control, As 4.8, 9.6, or 14.4, respectively, which exhibited a positive linear trend. There was evidence for trends between As dose and the number of litters displaying vertebral (po0.001) and calvarial (po0.01) abnormalities, components of the axial skeleton. Mean fetal weight of all As-treated groups was significantly less than in control. DISCUSSION: In our model, maternal oral treatment with As induced NTDs. It also significantly increased the frequency of axial skeletal anomalies in the offspring exposed in utero, and reduced mean fetal weight, without evidence of maternal toxicity. Birth Defects Res (Part B) 83: 40-47, 2008.

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“…[8] ARS has been widely used in the textile industry, but it also enjoys other applications such as staining bones in mammalian embryos [9] and skeletons in small invertebrate embryos. [10] However, ARS is a recalcitrant and durable dye that is both mutagenic and carcinogenic. ARS can form deeply colored metal complexes.…”

Section: Introductionmentioning

confidence: 99%