Drug Contamination ; Staining and Labeling ; methods

Humans ; Receptor, ErbB-2 ; Staining and Labeling

Biopsy ; Humans ; Kidney ; pathology ; Staining and Labeling ; methods

Docosahexaenoic acid (DHA) has many unique physiological functions such as promoting the development of brain and retina in infants. Therefore, it is widely applied to food, pharmacy, breeding and other industries. To obtain engineered strains of Aurantiochytrium limacinum SR21 suitable for industrial application with increased lipid and DHA production, we designed a simple, fast, accurate and high-throughput screening method based on Nile red staining of oil droplets. First, ultraviolet C (UVC) mutagenesis was used to generate a random mutant library of A. limacinum. Second, screening conditions were optimized including staining conditions of Nile red and the sorting criterion. Thereby, three putative high-lipid mutants (D03432, D05106 and D01521) were selected from the mutant library containing 3 648 mutated clones. The three mutants grew faster and produced higher amounts of lipids and DHA compared to wild type (WT). In 100 mL cultures, the lipid content of D03432 and D05106 mutants reached 64.74% and 63.13% of dry cell weight respectively, whereas the wild strain exhibited only 43.19%. DHA yield in these two mutants were even 2.26-fold and 2.37-fold higher than that of the wild strain. Experiment with 5 L fermentor further confirmed that D03432 and D05106 mutants had better performance than the wild strain on DHA yield (45.51% and 66.46% more than that of the wild strain, respectively), and demonstrated their high potential for industrial application. This work not only generated several high-DHA content mutants with high potential for industrial use, but also provided vital guidance for high-throughput screening of lipid hyper-accumulating mutants in other oil-producing microorganisms.
Bioreactors ; Docosahexaenoic Acids ; Mutagenesis ; Staining and Labeling ; Stramenopiles

Humans ; Methylene Blue ; Melanoma ; Staining and Labeling

OBJECTIVE: To explore the treatment conditions of acid decalcified specimens and improve the poor quality of sections and unclear structure of hematoxylin-eosin (HE) staining caused by the change in pH in tooth and hard tissue after acid decalcification. METHODS: A total of 20 cases of oral pathological specimens that contain hard tissues were decalcified and treated with routine treatment, concentrated ammonia water immersion treatment, and saturated lithium carbonate solution immersion treatment. The quality and HE staining effects of hard tissue sections treated with different methods were compared. RESULTS: Compared with routine treatment, lithium carbonate saturated solution treatment showed complete sections. Hematoxylin is strongly stained, the nucleus is clear, and the cytoplasm is bright. CONCLUSIONS Soaking acid decalcified specimens in lithium carbonate saturated solution before embedding in dehydration can neutralize the acidic environment of the tissue. The quality of sections and HE staining effect are improved and are suitable for the pretreatment of acid decalcified tissue samples of oral pathology.
Eosine Yellowish-(YS) ; Hematoxylin ; Staining and Labeling ; Tooth

Detergents ; Immunohistochemistry ; instrumentation ; Oxalic Acid ; Potassium Permanganate ; Staining and Labeling ; instrumentation

OBJECTIVETo study the histochemical staining in the diagnosis of osteosarcoma.

METHODSTo compare the effectiveness of picrosirius red, improved Ponceau trichrome and Masson trichrome staining methods on bone formation tissues in conventional osteosarcoma, paraosteal osteosarcoma, periosteal osteosarcoma, extraskeletal osteosarcoma, inflammatory myofibroblastic tumour, malignant fibrohistiocytoma, chondrosarcoma, fibrosis with ossification and calcification.

RESULTSWith modified Ponceau trichrome staining, bone formation tissues showed a homogenous, orange-red interblended with blue in color. From osteoid to mature bone the color changed from orange-red, light blue to dark blue. Fibrotic tissue was stained blue in color with striated appearance. Cartilage was not stained. Picrosirius red method gave bone formation tissues homogenous staining. Along with bone maturation, from osteoid tissue to mineralized bones, the color showed changes from light red, yellow, orange-red, red to dark purple. The cartilage demonstrated homogenous light red in color. Fibrous tissue stained red interblended with yellow in color, striated in shape. With Masson trichrome staining osteoid displayed pale blue and mineralized bone showed dark blue in color. Fibrotic tissue showed a striated blue staining.

CONCLUSIONThe modified Ponceau trichrome and Picrosirius red staining methods are better than Masson trichrome to demonstrate bone formation tissue in osteosarcoma. The former two methods could be also used in study on bone formation.

Eosine Yellowish-(YS) ; Frozen Sections ; Hematoxylin ; Humans ; Staining and Labeling

OBJECTIVETo observe the distribution of Annexin I and cytosolic phospholipose A2 (cPLA2) in the palatal process of dexamethasome teratogenerated mice.

METHODSCutting along the coronary plane of the heads of fetal mice to obtain palate in the 14th, 15th, 16th day of gestation by 10-week-old in-bred mice. The distribution of Annexin I and cPLA2 was checked by immunohistochemical staining.

RESULTSDuring the fusing of the palatal processes, the staining of Annexin I and cPLA2 was positive in the epithelial and mesenchymal cells of the palatal process, and the intensity of staining changed with time.

CONCLUSIONAnnexin I and cPLA2 can modulate the development of fetal palate to some extent, and they may be important mediators in the development of cleft palate induced by dexamethasone.