Groups of WT, WT/WT, CD23KO/WT, and WT/CD23KO mice were sensitized with OVA by mixing with alum and given intraperitoneally (i.p.). Sterile PBS was used as a placebo control. Mice were sensitized by i.p. injection of 100 µg OVA emulsified in 4 mg aluminum hydroxide (AlumInject; Pierce Chemical, Rockford, IL) in a total volume of 100 µl on day 1, followed by 100 µg OVA on day 7 and 14. Mice were then challenged via the airways with nebulized OVA (1% in PBS) or PBS for 30 min on day 21. On day 22, AHR was measured. On day 22 or 23, the mice were sacrificed to collect BAL fluid, serum, and lung tissue for histopathology staining. For B3B4 antibody blocking experiment, the OVA sensitized WT mice received 75 µg of B3B4 or rat IgG2a intranasally after light anaesthetization with avertin (i.p.) on day 20 and day 21 before challenge. One hour later after second treatment, the Ab treated mice were challenged. On day 22, AHR was measured and on day 23, the mice were sacrificed to collect BAL fluid, serum, and lung tissue for histopathology staining. BAL fluid was subjected to cytospin, staining with modified Wright-Giemsa stain (Sigma), and differential counting of cells.
Modified wright giemsa stain
Manufactured by Merck Group
Sourced in Germany
The Modified Wright-Giemsa stain is a laboratory reagent used for the differential staining of blood cells. It combines the properties of the Wright stain and the Giemsa stain to enable the visualization and identification of various types of blood cells, including red blood cells, white blood cells, and platelets.
Automatically generated - may contain errors
Lab products found in correlation
Aluminject, by Thermo Fisher Scientific (2 mentions)
P akt, by Cell Signaling Technology (1 mentions)
P gsk3β, by Cell Signaling Technology (1 mentions)
Notch1, by Santa Cruz Biotechnology (1 mentions)
Gsk3β, by Cell Signaling Technology (1 mentions)
P27kip1, by Santa Cruz Biotechnology (1 mentions)
Mts assay, by Promega (1 mentions)
Optical microscope, by Nikon (1 mentions)
Ricin, by Merck Group (1 mentions)
Cx41 microscope, by Olympus (1 mentions)
Microtainer, by BD (1 mentions)
Hemavet hv950, by Drew Scientific (1 mentions)
13 protocols using modified wright giemsa stain
1
Allergen-Induced Airway Inflammation Study
2
Allergen-Induced Airway Inflammation Study
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Groups of WT, WT/WT, CD23KO/WT, and WT/CD23KO mice were sensitized with OVA by mixing with alum and given intraperitoneally (i.p.). Sterile PBS was used as a placebo control. Mice were sensitized by i.p. injection of 100 µg OVA emulsified in 4 mg aluminum hydroxide (AlumInject; Pierce Chemical, Rockford, IL) in a total volume of 100 µl on day 1, followed by 100 µg OVA on day 7 and 14. Mice were then challenged via the airways with nebulized OVA (1% in PBS) or PBS for 30 min on day 21. On day 22, AHR was measured. On day 22 or 23, the mice were sacrificed to collect BAL fluid, serum, and lung tissue for histopathology staining. For B3B4 antibody blocking experiment, the OVA sensitized WT mice received 75 µg of B3B4 or rat IgG2a intranasally after light anaesthetization with avertin (i.p.) on day 20 and day 21 before challenge. One hour later after second treatment, the Ab treated mice were challenged. On day 22, AHR was measured and on day 23, the mice were sacrificed to collect BAL fluid, serum, and lung tissue for histopathology staining. BAL fluid was subjected to cytospin, staining with modified Wright-Giemsa stain (Sigma), and differential counting of cells.
3
Bronchoalveolar Lavage Cell Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Offspring were euthanized 48 h post-challenge and lungs were lavaged with 0.8 mL ice-cold 1X PBS supplemented with 0.2 mM EDTA (bronchoalveolar lavage fluid or BAL). An aliquot of BAL was applied to a hemocytometer and total cell numbers were calculated. An aliquot of BAL was then applied to slides using the Shandon Cytospin, fixed with methanol, and stained with modified Wright-Giemsa stain (Sigma-Aldrich). Two individuals, blinded to treatment, counted 200 cells/slide and identified cells as macrophages, eosinophils, neutrophils or lymphocytes. Total cell numbers were then extrapolated for each cell type. The remaining BAL was centrifuged and the supernatant was stored at −80°C prior to cytokine analysis.
4
Asthma and Airway Inflammation Modulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ovalbumin (OVA), methacholine, dexamethasone (DEX), tangeretin, and modified Wright-Giemsa stain were from Sigma-Aldrich (USA). ELISA (Enzyme-linked immunosorbent assay) kits for IL-4, IL-5, and IL-13 were obtained from Biolegend (USA). Antibodies against cyclinA, cyclinD1, Akt, p-Akt, GSK-3β, p- GSK-3β, phosphatise, and PTEN were from Cell Signaling Technology (USA). Mammalian target of rapamycin (mTORc1), Jagged 1, Jagged 2, Notch 1, Notch receptor intracellular domain (NICD), and p27kip1 antibodies were from Santa Cruz Biotechnology (USA). FITC-labelled anti-human CD4, phycoerythrin (PE), and anti-mouse IL-17A were from eBioscience Co. (USA). All other analytical-grade chemicals were from Sigma-Aldrich unless noted otherwise.
5
Morphology and Phagocytic Assay of Primary PAMs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The morphological characteristics of the cells were observed by staining with modified Wright–Giemsa stain (Sigma-Aldrich; Merck KGA, Darmstadt, Germany) according to the manufacturer’s protocol.
The phagocytic activity of primary PAMs was determined by adding chicken erythrocytes to a final concentration of 104/mL and incubated for 12 hours. The degree of phagocytosis was then observed under an optical microscope (Nikon Corporation, Tokyo, Japan).
The half maximal inhibitory concentration (IC50) of ricin (Sigma-Aldrich, Corp., St. Louis, MO, USA) was tested by MTS assay (Promega Corporation, Madison, WI, USA). In brief, the cells were grown in 96-well plates and treated with serially diluted ricin (0–105 ng/mL) for 12 hours, followed by the addition of 20 µL MTS to each well. After incubation for 2 hours at 37°C, the absorption was determined at 490 nm. All experiments were performed in triplicate.
The phagocytic activity of primary PAMs was determined by adding chicken erythrocytes to a final concentration of 104/mL and incubated for 12 hours. The degree of phagocytosis was then observed under an optical microscope (Nikon Corporation, Tokyo, Japan).
The half maximal inhibitory concentration (IC50) of ricin (Sigma-Aldrich, Corp., St. Louis, MO, USA) was tested by MTS assay (Promega Corporation, Madison, WI, USA). In brief, the cells were grown in 96-well plates and treated with serially diluted ricin (0–105 ng/mL) for 12 hours, followed by the addition of 20 µL MTS to each well. After incubation for 2 hours at 37°C, the absorption was determined at 490 nm. All experiments were performed in triplicate.
6
Cell Preparation and Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were placed on slides by centrifugation at 700 rpm for 5 min (CytoSpin 3 Cytocentrifuge, Shandon). Slides were air-dried prior to staining. For morphological analysis, slides were stained with modified Wright–Giemsa stain (Sigma-Aldrich) according to the manufacturer’s protocol.
7
Bronchoalveolar Lavage Fluid Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BAL was performed with a total volume of 1 ml PBS containing 100 μM EDTA. BALF was collected in Eppendorf tubes on ice, with aliquots made for flow cytometry, cytospin differential counts, and an aliquot transferred to -80°C for multiplex cytokine analysis. Cytospin preparations of cells were stained with modified Wright-Giemsa stain (Sigma-Aldrich).
8
Bronchoalveolar Lavage Fluid Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BAL was performed with a total volume of 1 mL PBS containing 100 μM EDTA. BALF was collected in Eppendorf tubes on ice, with aliquots made for flow cytometry, cytospin differential counts, and an aliquot transferred to −80 °C for multiplex cytokine analysis. Cytospin preparations of cells were stained with modified Wright-Giemsa stain (Sigma-Aldrich, St. Louis, MO).
9
Bronchoalveolar Lavage Fluid Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BAL was performed with a total volume of 1 ml PBS containing 100 μM EDTA. BALF was collected in Eppendorf tubes on ice, with aliquots made for flow cytometry, cytospin differential counts, and an aliquot transferred to -80°C for multiplex cytokine analysis. Cytospin preparations of cells were stained with modified Wright-Giemsa stain (Sigma-Aldrich). Murine TRAP5 was measured in BALF using ELISA (Abcam).
10
Leukocyte and Erythrocyte Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Differential leukocyte counts and frequency of immature erythrocytes were analysed from blood smears stained with Modified Wright-Giemsa Stain (product no. WG128, Sigma-Aldrich) and scanned using an Olympus CX-41 microscope (Olympus, Japan) under 1000× magnification. The proportion of lymphocytes and heterophils was calculated from a sample of 120–160 leukocytes per smear (see [50 ]). Repeatability of the estimate was rlymphocytes = 0.93, rheterophils = 0.90, n = 10, p < 0.001; [51 ]).
The differential count of immature erythrocytes was estimated from 5–10 randomly chosen monolayer fields photographed at 100× objective magnification (ca. 800–1500 cells). Immature erythrocytes were manually counted from the photographs using ImageJ software 1.48 [52 ]. Repeatability of the measurement was assessed as r = 0.97 (n = 15, p < 0.001).
The differential count of immature erythrocytes was estimated from 5–10 randomly chosen monolayer fields photographed at 100× objective magnification (ca. 800–1500 cells). Immature erythrocytes were manually counted from the photographs using ImageJ software 1.48 [52 ]. Repeatability of the measurement was assessed as r = 0.97 (n = 15, p < 0.001).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!