Application Carmine has been used for the staining of mammary glands. The inguinal mammary glands from 3-, 4.5-, 6- and 10-week-old female C57Bl6/J mice (Harlan Laboratories) were fixed overnight in Carnoy’s fixative. Both ES and BI performed carmine staining of whole mounts, cultured tumor-derived mammary cells, and performed western blotting on lysates obtained from NSD3-derived mammary tumor cells. Whole-mount staining of mammary glands. LS is the breast cancer pathologist for the Hollings Cancer Center and along with JM … Whole mount Carmine staining of #4 mammary glands in 8 week old males from control (I) and Ift88 Orpk (L; Bl/6 shown). Male Bl/6 and Balb/c mice do not generally develop a mammary ductal tree. Rats were sacrificed after completions of study and forth mammary gland tissues were excised and fixed for a minimum of 2 days in Carnoy’s fixative solution followed by staining with carmine solution (1 g carmine and 2.5 g aluminum potassium sulfate in 500 ml water). The glands were then immersed in Carnoy’s fixative (3:1, ethanol:glacial acetic acid) overnight, washed in 70% ethanol for 15 minutes, washed under running tap water for 5 minutes, drained and then stained overnight with Alum Carmine stain [2 g/l carmine dye (C6152, Sigma, St Louis, MO) and 5 g/l aluminum potassium sulfate in distilled water]. BBAP-2 restored the morphological architecture when screened through carmine staining, haematoxylin and eosin staining, and scanning electron microscopy. Dissected fourth and fifth mammary glands were spread on a glass slide. Quantification indicating decreased numbers of tertiary branches in Pdgfra-CreER; β-cat fl/fl mice (J). In vivo study revealed the morphological changes of mammary gland tissue when scrutinized using carmine staining, hematoxylin and eosin staining and scanning electron microscopy. 2.3 Whole‐mount mammary gland carmine staining. This pigment stains the mammary epithelia an intense pink/red colour and relies on the solvent xylene or methyl salicylate for subsequent optical transparency [ 29 , 30 ]. Suitability of Tissue Sections for Other AssaysThe initial carmine staining of whole mounts does not appear to interfere with histochemical or immunohistochemical (IHC) stains. Abnormalities within the mammary gland are usually not visible until after the gland has been defatted in xylene or has been carmine-stained. 3.2.1 Mammary gland carmine alum staining 75 3.2.2 Immunofluorescent immunostaining of mouse mammary gland sections 76 3.2.3 Lymphatic vessel quantification 76 3.2.4 Whole mount immunofluorescent immunostaining of mouse mammary glands 77 3.3 RESULTS 78 3.3.1 The ductal epithelial network is extensively remodelled during postnatal mouse mammary gland morphogenesis 78 . These stains include the … The fixed glands were hydrated with 90, 70, 35 and 15% ethyl alcohol and then rinsed with water for 5 min. Carmine staining. Knockout efficiency was validated by qPCR (K). Mammary whole‐mount analysis was performed as described previously (Plante … The glands were fixed in a mixture of 6 : 3 : 1 methanol (Sigma, Zwijndrecht, The Netherlands) : 1,1,1‐trichloroethane (Sigma) : acetic acid (Sigma) for 4 h and processed as whole mounts, which were stained overnight with carmine aluminium staining solution [2 g/l carmine … Whole mount carmine alum staining. The mammary gland of Pdgfra-CreER;β-cat fl/fl mice had fewer branches compared with that of Ctrl mice (I). Mammary glands were harvested and fixed in cold PFA for 2 hr, washed with PBS for 3 times (10 min each), followed by staining in carmine solution overnight. The effect of exogenous hormone treatment on mammary gland morphogenesis. (A-F) Carmine alum-stained whole-mounts (A-C) and F4/80 staining (D-F) from ovariectomised unstimulated mice (A,D), mice administered exogenous estradiol and progesterone (B,E) and estradiol and progesterone followed by mifepristone (C,F). Mammary gland morphology in MTB mice during development. The fixed mammary glands were hydrated using descending ethanol gradient and stained in carmine alum solution for overnight. The #4 inguinal mammary gland was collected to determine the area where mammary glands were developed in … For mammary gland branching analysis, 4th inguinal mammary fat pads were prepared as whole mount samples and carmine alum staining was performed according to the protocol as previously described . Scale bar, 1 mm. In presence of aluminum chloride, it is used for the staining of glycogen. Finally, the glands are stained in the above wash solution supplemented with 30 mM potassium … 5A) composed of many … VitroView™ Mammary Gland Whole Mount Stain Kit is very useful for the study of mammary gland development. Mammary glands were excised, spread on glass slides, and fixed in Carnoy’s fixative (60% ethanol, 30% chloroform, and 10% acetic acid) at room temperature for 6 h. After washing in 70% ethanol and distilled water, mammary glands were stained overnight at room temperature in carmine alum solution (0.2% carmine and 0.5% aluminum potassium sulfate). Whole mount carmine alum staining Mammary glands were excised, spread on glass slides, and fixed in Carnoy’s fixative (60% ethanol, 30% chloroform, and 10% acetic acid) at room temperature for 6 h. After washing in 70% ethanol and distilled water, mammary glands were stained overnight at room temperature in carmine alum solution (0.2% carmine and 0.5% alu-minum potassium sulfate). Angiotensin II (AngII), a major vasopressor hormone, has key roles in regulation of blood pressure, whereas a recent work revealed the association between AngII and mammary gland … This product contains carmine and aluminum potassium sulfate in water. Unlike mammary tissue that was originally prepared for histological sectioning, a whole mount removed from the glass for paraffin embedding will be extremely thin and fragile. To investigate organ-level changes, we visualized mammary ducts with carmine alum staining at day 30 (Figure 3D). Carmine alum staining. … Carmine Alum is used for the whole mount staining of mouse mammary gland. A: Microscopic examination of rat mammary gland tissue through carmine staining (A1-G1) and H&E staining (A2-G2). Mammary glands from control and TM-treated Her2/neu and FVB mice were isolated and processed for whole-mount analysis using carmine red staining. A1 and G1 shows that carmine staining of mammary gland tissue of … Whole inguinal mammary glands from 10 mice of each genotype were dissected out, mounted and spread onto superfrost glass slides, fixed overnight in Carnoy’s fixative (ethanol, chloroform, glacial acetic acid 6:3:1) followed by rehydration and overnight staining with carmine red stain [1 g carmine red dye (Sigma), 2.5 g potassium alum in 500 ml water]. Reconstituted mammary glands were dissected and stretched on a glass slide. Columns, mean number of secondary branches per field; bars, … 4, B and C). Alternatively, glands were fixed in 4%paraformaldehyde (PFA) in PBS for 1 hour, washed three times in rinse buffer(2 mM MgCl 2, 0.2% sodium deoxycholate, 0.2% NP40 in PBS) and stained overnight at room temperature in 5-bromo-4 … The contralateral inguinal mammary gland in each mouse was used as the sham control. The fourth inguinal mammary glands were selected for the study because they contain a centrally located lymph node that can guide the site of inoculation, and because they have been widely studied [18-20]. BBAP-1 treatment produced a marked effect on histopathological and morphological features when scrutinized against N-methyl-N-nitrosourea induced mammary gland carcinoma.

(A, B) Carmine whole mount staining of the fourth mammary gland isolated from EE and SE mice. (n = 6; scale bars; upper panels: 5 mm, lower panels: 250 µm). Full size image. The glands were then washed 3 × 30 min. For analysis of whole‐mount morphology (A), inguinal mammary glands were spread on slides, fixed, and subjected to carmine staining . Arrows indicate F4/80-stained macrophages. Currently, carmine is the most prevalent histochemical stain for assessing mammary gland morphogenesis in whole mount.
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