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Diff quik

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Diff-Quik is a rapid, three-step staining method used for the differential staining of blood smears and other cytological specimens. It provides a quick and reliable way to obtain detailed information about the cellular composition of a sample.

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Lab products found in correlation

Nucleocounter, by ChemoMetec (3 mentions) Transwell insert, by BD (2 mentions) Ca2 and mg2 free pbs, by Thermo Fisher Scientific (2 mentions) Cytospin, by Hanil (2 mentions) Nintedanib, by Cayman Chemical (2 mentions) Obe022, by Abbvie (2 mentions) Bay6872, by Abbvie (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Thinprep, by Hologic (1 mentions) Nucleocounter yc 100, by ChemoMetec (1 mentions) Sdf 1, by R&D Systems (1 mentions) Vegf165, by Merck Group (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Ethylene diamine tetraacetic acid (edta), by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Protein synthesis assay kit, by Abcam (1 mentions) Cytospin 4 cytocentrifuge, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) Lsrfortessa, by BD (1 mentions)

Close spelling variants of this product

Diff-Quik Staining Kit (9 citations) Diff-Quik staining (2 citations) Diff‐Quik stain (2 citations) Diff Quik solution (2 citations)

33 protocols using diff quik

1

Cell Migration and Invasion Assay

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To assess cell migration, cells (1 × 105) were seeded atop an 8-μm microporous membrane located within the upper compartment of a transwell insert (BD Biosciences), and incubated at 37°C for 24 hours. After incubation, migrated cells passing through the 8-μm pore filter were fixed, stained with Diff-Quik (Thermo Fisher Scientific), and enumerated using ImageJ software. All experiments were completed in triplicate and five fields/well were counted. To assess invasion, an 8-μm microporous membrane located within the upper compartment of a transwell insert (BD Biosciences) was coated with 20 μg of rat tail type I collagen in sodium carbonate, pH 9.6 overnight at 4°C, washed with PBS, and air dried. Cells were seeded atop the filter and the apparatus incubated at 37°C for 24 hours. After incubation, invaded cells passing through the collagen and 8mm pore filter were fixed, stained with Diff-Quik (Thermo Fisher Scientific), and enumerated using ImageJ software. All experiments were completed in triplicate and five fields/well were counted.
Asem M., Young A.M., Oyama C., De La Zerda A.C., Liu Y., Yang J., Hilliard T.S., Johnson J., Harper E.I., Guldner I., Zhang S., Page-Mayberry T., Kaliney W.J, & Stack M.S. (2020). Host Wnt5a Potentiates Microenvironmental Regulation of Ovarian Cancer Metastasis. Cancer research, 80(5), 1156-1170.
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2

Three-Dimensional Ovarian Cancer Migration Assay

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A three-dimensional collagen type I matrix was prepared (previously described, (Moss et al. 2009b (link))) with or without fibroblasts atop an 8μm microporous membrane within a transwell insert (BD Biosciences, San Diego, Ca) and overlaid with LP9 cells (Lengyel et al. 2013 ). The LP9 were allowed to grow to confluence, forming a tight monolayer. CellTracker™ Green-labelled OVCA433 cells were seeded atop the live monolayer and the co-culture was incubated at 37°C for 48 hours in a 1:1 ratio of complete media for each cell type. Migrated cells passing through the 3-dimensional culture and the 8μm pore filter were fixed and stained with Diff-Quik (Fisher Scientific, Pittsburgh, PA) and enumerated. All experiments were completed in triplicate and five fields/well were counted. Results show mean +/− SEM. Student’s t-test was used to determine p values (Sigmaplot).
Bruney L., Conley K.C., Moss N.M., Liu Y, & Stack M.S. (2014). Membrane-type I Matrix Metalloproteinase-Dependent Ectodomain Shedding of Mucin16/CA-125 on Ovarian Cancer Cells Modulates Adhesion and Invasion of Peritoneal Mesothelium. Biological chemistry, 395(10), 1221-1231.
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3

Cytological Specimen Preparation and Molecular Testing

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The cytological specimen preparations were conducted according to a standard specimen processing protocol in our laboratory. The cases included FNA specimens obtained under image guidance with a cytopathologist present for adequacy assessment and pleural fluid specimens obtained by thoracentesis. The percentage of tumor cells more than 5% or over 500 tumor cells were used for DNA extraction and 100 tumor cells for ALK FISH analysis. The algorithm used for molecular testing in our study was depicted in Supplementary Figure S1. Each case had air-dried slides stained with Diff-Quik (DQ stain, Protocol Hema 3; Fisher Scientific, Kalamazoo, MI) and additional slides fixed in 95% alcohol for Papanicolaou staining. Fluid specimens also had a ThinPrep (Hologic, Marlborough, MA) slide prepared.
Li W., Zhang Z., Guo L., Qiu T., Ling Y., Cao J., Guo H., Zhao H., Li L, & Ying J. (2015). Assessment of cytology based molecular analysis to guide targeted therapy in advanced non-small-cell lung cancer. Oncotarget, 7(7), 8332-8340.
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4

BALF Cell Differential Analysis

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BALF was centrifuged at 200×g for 5 minutes, and supernatant was removed for cytokine measurement. Pelleted cells were resuspended and counted by NucleoCounter YC-100 (ChemoMetec, Allerød, Denmark) automated cell number counting. 100 µl of cell suspension was then loaded onto a glass slide using a disposable sample funnel and cytocentrifuged at 10×g for 3 minutes in a Shandon Cytospin 2 centrifuge. Slides were air dried for 20 minutes, fixed in methanol for 20 minutes, stained with Diff Quik (Fisher Scientific, Loughborough, UK), and mounted in DPX Mountant (Fluka BioChemika/Sigma Aldrich, UK). Differential counts for neutrophils and monocytes were then performed by light microscopy at 20× magnification using an EVOS FL microscope (Peqlab, Sarisbury Green, UK).
Beaumont P.E., McHugh B., Gwyer Findlay E., Mackellar A., Mackenzie K.J., Gallo R.L., Govan J.R., Simpson A.J, & Davidson D.J. (2014). Cathelicidin Host Defence Peptide Augments Clearance of Pulmonary Pseudomonas aeruginosa Infection by Its Influence on Neutrophil Function In Vivo. PLoS ONE, 9(6), e99029.
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5

BAL and Lung Tissue Collection

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Lung tissue and BAL samples were obtained as previously described [34] (link), [35] (link). Briefly, mice were anesthetized and a small-caliber tubing was inserted into the trachea. Two successive volumes of 1 ml of PBS were instilled, aspirated and pooled. BAL samples were centrifuged at 4000 rpm, and supernatants were stored at −80°C until evaluation. Cells in 100 µl aliquots were counted by trypan blue staining. A total of 100,000 viable BAL cells were centrifuged in Cytospin 3 (Thermo Shandon Ltd, Runcorn, UK). Cell differentiation was determined by Diff-quik (Fisher Scientific Co., Newark, DE). The lung was perfused with cold PBS. The whole lung was either excised for RNA and protein analyses or inflated with neutral buffered formalin for histology.
Fang P., Zhou L., Zhou Y., Kolls J.K., Zheng T, & Zhu Z. (2014). Immune Modulatory Effects of IL-22 on Allergen-Induced Pulmonary Inflammation. PLoS ONE, 9(9), e107454.
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6

Murine Lung Tissue and Fluid Analysis

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When Flexivent was not performed, mice were anesthetized with isoflurane and euthanized by exsanguination. Lungs were removed from the chest cavity and the trachea canulated. The right lung was tied using suture strings, snap frozen in liquid nitrogen, and kept at − 80 °C. Bronchoalveolar lavage (BAL) was performed on the left lung by lavaging the lungs twice with 250 μl of PBS. Following BAL, the left lung was inflated with 10% formalin and embedded in paraffin for histological assessment. BAL total cell concentration was determined using a hemocytometer. BAL cells were pelleted at 800 x g, the cell-free BAL fluid (BALF) was collected and stored at − 80 °C. The cell pellet was resuspended in PBS to perform cytospins. Cytospins were stained (Diff-Quik; Fisher Scientific, Ottawa, ON, Canada) and differential counts performed using the Image J software by counting at least 300 cells per cytospin. Blood was collected, left to coagulate at 37 °C, and the serum separated by centrifugation and stored at − 80 °C.
Talbot M., Hamel-Auger M., Beaulieu M.J., Gazzola M., Lechasseur A., Aubin S., Paré M.È., Marsolais D., Bossé Y, & Morissette M.C. (2018). Impact of immunization against OxLDL on the pulmonary response to cigarette smoke exposure in mice. Respiratory Research, 19, 131.
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7

Boyden Chamber Assay for Cell Migration

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Cell migration was assessed using a modified-Boyden chamber assay. CACs were detached from culture slides, washed with PBS and resuspended at a density of 5×105/ml in migration medium (EBM-2 medium containing 0.5% BSA). 500 µl (2.5×105) of cells were then added into the top chamber of the modified Boyden chamber apparatus (BD Biosciences, 8 µm pores). The chemoattractants VEGF165 (Sigma) or stromal cell derived factor 1 (SDF1, R&D Systems), used to promote migration, were prepared with the migration medium at concentrations of 50 ng/ml and 100 ng/ml, respectively. 500 µl of chemoattractant or migration medium alone was added to the lower chamber. Following a 5 hour incubation period at 37°C migratory, cells present on the underside of the insert were fixed and stained using Diff Quik (Fisher Scientific) and visualized by light microscopy. Images were acquired for 5 randomly selected fields and the mean number of cells from these fields was determined. Data are presented as fold change in cell migration towards chemoattractant compared to the respective control basal migration.
Zucco L., Zhang Q., Kuliszewski M.A., Kandic I., Faughnan M.E., Stewart D.J, & Kutryk M.J. (2014). Circulating Angiogenic Cell Dysfunction in Patients with Hereditary Hemorrhagic Telangiectasia. PLoS ONE, 9(2), e89927.
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8

Bronchoalveolar Lavage Fluid Analysis

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After 4 hours of reperfusion, a clamp was placed across the right hilum and the left lung lavaged with sterile saline. The recovered fluid was centrifuged at 1,800 rpm for 10 minutes at 4°C. The supernatant was collected from each sample and assessed for cytokines. The cell pellet was re-suspended and lysed for red blood cells. Cells were additionally washed at 1,200 rpm for 10 minutes and re-suspended in 1 mL of PBS. Total cell counts were assessed using a hemacytometer.
BAL samples were centrifuged at 1200 rpm for 10 minutes at room temperature and cells resuspended in 500 uL of PBS. Cytospins were prepared from cells isolated from BAL and stained in Diff-Quik (Fisher Scientific, Pittsburg, PA) to assess for neutrophils, macrophages, and lymphocytes.
Hwang B., Liles W.C., Waworuntu R, & Mulligan M.S. (2015). Pretreatment with Bone Marrow Derived Mesenchymal Stromal Cell Conditioned Media Confers Pulmonary Ischemic Tolerance. The Journal of thoracic and cardiovascular surgery, 151(3), 841-849.
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9

Macrophage-Leishmania Infection Protocol

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Bone marrow derived macrophages (BMMs) were cultured at 37°C, 5% CO2 in RP-10 (RPMI with 10% heat-inactivated fetal calf serum, 2 mM L-glutamine, 100 U of penicillin/ml, and 50 μg of streptomycin/ml (GIBCO, Carlsbad, CA) containing 20% L929 cell culture supernatant (American Tissue Type Collection, Manassas, VA) as a source of macrophage colony-stimulating factor. After 7 to 9 days, differentiated adherent BMMs were detached from Petri dishes with 2.5 mg trypsin/ml plus 1 mM EDTA (GIBCO), and cultivated overnight on glass coverslips prior to use.
Metacyclic L. infantum, attenuated L5 promastigotes, hamster-derived L. infantum amastigotes, virulent S. typhimurium or GFP-latex beads were opsonized with 5% C5-deficient A/J mouse serum for 30 min at 37°C. Five x 105 BMMs adherent to glass coverslips were infected with L. infantum parasites as described. Briefly, L5 or metacyclic L. infantum promastigotes were used at a multiplicity of infection [MOI] of 5:1; amastigotes were used at a MOI of 2:1. Infections were synchronized by centrifugation (3 min, 330 x g, 4 °C), and macrophages incubated at 5% CO2, 37°C. After 30 minutes, extracellular particles were removed by rinsing and macrophages were returned to 37°C, 5% CO2. At specified time points, triplicate coverslips were fixed and stained with Wright Giemsa (Diff-Quik, Fisher Scientific) [26 (link), 28 (link)].
Dixit U.G., Rodríguez N.E., Polando R., McDowell M.A, & Wilson M.E. (2020). Complement receptor 3 mediates ruffle-like, actin-rich aggregates during phagocytosis of Leishmania infantum metacyclics. Experimental parasitology, 220, 107968.
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10

Flow Cytometry and Diff-Quik Staining

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Cells were analyzed by flow cytometry as described by Olschok et al. (2021) and Toledo et al. (2021) (link) (Figure S4B; Table S1) and analyzed with FACS Canto II or LSR Fortessa and FlowJo software (all BD Bioscience, Franklin Lakes, NJ). For the analysis of protein biosynthesis with flow cytometry, the protein synthesis assay kit (ab239725, Abcam, Cambridge, UK) was used.
Diff-Quik staining cells were centrifuged onto glass slides in the Cytospin 4 cytocentrifuge (Thermo Fisher Scientific). Cells were fixed with methanol, stained with Diff-Quik (Medion Diagnostics, Düdingen, Switzerland), and mounted with Entellan (Merck, Rahway, NJ).
Flosdorf N., Böhnke J., de Toledo M.A., Lutterbach N., Lerma V.G., Graßhoff M., Olschok K., Gupta S., Tharmapalan V., Schmitz S., Götz K., Schüler H.M., Maurer A., Sontag S., Küstermann C., Seré K., Wagner W., Costa I.G., Brümmendorf T.H., Koschmieder S., Chatain N., Castilho M., Schneider R.K, & Zenke M. (2024). Proinflammatory phenotype of iPS cell-derived JAK2 V617F megakaryocytes induces fibrosis in 3D in vitro bone marrow niche. Stem Cell Reports, 19(2), 224-238.
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