Diff quik stain

Manufactured by Sysmex

Sourced in Japan

Diff-Quik stain is a fast and reliable staining solution used for the differentiation and identification of various cell types in blood smears and other cytological preparations. It provides high-quality staining of cellular components, enabling efficient analysis and diagnosis.

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Lab products found in correlation

Matrigel, by BD (8 mentions) Streptomycin, by Thermo Fisher Scientific (5 mentions) Penicillin, by Thermo Fisher Scientific (4 mentions) Transwell chamber, by Corning (4 mentions) Biocoat matrigel invasion chamber, by Corning (4 mentions) 24 well modified boyden chambers transwell chamber, by BD (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Shandon cytospin 4, by Thermo Fisher Scientific (3 mentions) Transwell chamber, by BD (2 mentions) Cytopro 7620, by Wescor (2 mentions) Transwell chamber, by Thermo Fisher Scientific (2 mentions) Biocoat matrigeltm invasion chamber kit, by BD (2 mentions) Biocoat matrigel invasion chambers with 8.0 μm pet membrane, by Corning (2 mentions) Biocoat matrigel invasion chamber, by BD (2 mentions) Percoll, by Thermo Fisher Scientific (1 mentions) Histopaque 1119, by Merck Group (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Percoll, by GE Healthcare (1 mentions) Celltac hematology analyzer, by Nihon Kohden (1 mentions) Falcon cell culture insert, by Corning (1 mentions)

37 protocols using diff quik stain

Cytological Analysis for Endometritis

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Samples for cytological analysis were prepared by rolling the cytobrushes on sterile glass slides and immediately fixing the slides on-site using a
cytofixative agent (Cytokeep II, Alfresa Pharma, Osaka, Japan). The slides were then transported to the laboratory within 3 h and stained using the Diff-Quik
stain (Sysmex, Kobe, Japan) for 20 sec. The percentage of neutrophils (PMN) was evaluated as described by Ghanem et al. [17 (link)]. The threshold values for PMN indicating subclinical endometritis were ≥ 6% at W5 and ≥ 4% at W7 [20 (link)].

GHANEM M.E., TEZUKA E., SASAKI K., TAKAHASHI M., YAMAGISHI N., IZAIKE Y, & OSAWA T. (2016). Correlation of blood metabolite concentrations and body condition scores with persistent postpartum uterine bacterial infection in dairy cows. The Journal of Reproduction and Development, 62(5), 457-463.

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Quantifying HCC Cell Invasion

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We evaluated the invasion ability of cells using 24-well Corning BioCoat Matrigel Invasion Chambers (Corning). The lower chamber was filled with medium supplemented with 3% FBS containing CnP (0.3 mg/mL), CAF-CM, or CnP-treated CAF-CM, and HCC cell lines in DMEM (serum-free) were seeded into the upper chamber. After 48 hours, Diff-Quik stain (Sysmex) were used for fixity and staining the cells. We evaluated five randomly selected fields under a microscope and counted the cells that invaded and migrated through the membrane.

Yamanaka T., Harimoto N., Yokobori T., Muranushi R., Hoshino K., Hagiwara K., Gantumur D., Handa T., Ishii N., Tsukagoshi M., Igarashi T., Watanabe A., Kubo N., Araki K., Umezawa K, & Shirabe K. (2021). Conophylline Inhibits Hepatocellular Carcinoma by Inhibiting Activated Cancer-associated Fibroblasts Through Suppression of G Protein-coupled Receptor 68. Molecular cancer therapeutics, 20(6).

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Murine Neutrophil Isolation and Characterization

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For each mouse, 20 μL of blood was analyzed to count the number of white blood cells (WBCs) using the Celltac hematology analyzer (MEK-6308; Nihon Kohden, Tokyo, Japan).
For neutrophil isolation, blood was collected from each mouse followed by separation with centrifugation for 20 minutes at 800 g using Histopaque-1119 (Sigma-Aldrich, St. Louis, MO, USA). The neutrophil-rich phase was collected, washed in PBS, and separated by discontinuous density-gradient centrifugation in Percoll (GE Healthcare, Buckinghamshire, UK), as previously described [22 (link)]. Subsequently, the neutrophils were collected from the 70–75% layer of the Percoll gradient and washed with PBS; after hypotonic lysis with 0.2% and 1.6% NaCl solutions to remove residual erythrocytes, the cells were resuspended in RPMI-1640 (Invitrogen, Waltham, MA, USA). Purity and viability were routinely assessed using Diff-Quik stain (Sysmex, Kobe, Japan) and trypan blue stain (Wako Pure Chemical Industries, Osaka, Japan), respectively, under a microscope.

Hamaguchi S., Akeda Y., Yamamoto N., Seki M., Yamamoto K., Oishi K, & Tomono K. (2015). Origin of Circulating Free DNA in Sepsis: Analysis of the CLP Mouse Model. Mediators of Inflammation, 2015, 614518.

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Quantitative Cell Invasion Assay

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Invasion assays were performed in modified Boyden chambers (Chemotaxis 8μ; Kurabo); cells (2 × 10⁵) in 300 μL of serum‐free RPMI were loaded into the upper well. RPMI (500 μL) with 10% fetal bovine serum was added to the lower well. After a 24‐hours incubation, cells in the upper chambers were removed with a cotton swab, and the filter was fixed and stained with Diff‐Quik stain™ (Sysmex). Three randomly selected fields in each membrane were photographed under an Olympus C‐5060 light microscope (Olympus) with a 10 × objective.

Moritake H., Saito Y., Sawa D., Sameshima N., Yamada A., Kinoshita M., Kamimura S., Konomoto T, & Nunoi H. (2019). TAE226, a dual inhibitor of focal adhesion kinase and insulin‐like growth factor‐I receptor, is effective for Ewing sarcoma. Cancer Medicine, 8(18), 7809-7821.

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Assessing Cell Migration via Wound Scratch and Transwell Assays

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Wound scratch and Transwell migration assays were conducted as previously described^{10 (link)}. A549 cells were pretreated with dexamethasone (2.5 μM) or fluticasone propionate (2.5 μM) for 1 hour and then stimulated with TGF-β1 (5 ng/ml). For the wound scratch migration assays, cells were photographed with a digital camera (Olympus BX51; Olympus, Tokyo, Japan) 48 hours after the scratch. Wound closure of the cells that crossed into the scratch area as compared to zero time was calculated by ImageJ software (National Institutes of Health, Bethesda, MD, USA). Graphs represent the percentage wound closure. For the Transwell migration assays, A549 cells from the migrated side were stained with Diff-Quik stain (Sysmex, Kobe, Japan). Images were acquired under a microscope at 200× magnification (Olympus BX51; Olympus), and stained cells were counted in five high-power fields (HPFs) to determine the average cell number migrated per HPF.

Yang H.W., Lee S.A., Shin J.M., Park I.H, & Lee H.M. (2017). Glucocorticoids ameliorate TGF-β1-mediated epithelial-to-mesenchymal transition of airway epithelium through MAPK and Snail/Slug signaling pathways. Scientific Reports, 7, 3486.

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Transwell Migration and Invasion Assay Protocol

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Transwell migration and invasion assays were carried out in 24-well modified Boyden chambers (transwell-chamber, BD Transduction, Franklin Lakes, NJ). The upper surface of the 6.4 mm diameter filters with 8-μm pores was pre-coated with (invasion assay) or without (migration assay) Matrigel (BD Transduction). The siRNA transfectants (2 × 10⁴ cells per well) were transferred into the upper chamber. Following 48 h of incubation, migrated or invasive cells on the lower surface of filters were fixed and stained with the Diff-Quik stain (Sysmex, Kobe, Japan), and stained cell nuclei were counted directly in triplicate. We assessed the invasive potential by calculating the ratio of the percentage invasion through the Matrigel-coated filters relative to migration through the uncoated filters of test cells over that in the control counterparts [33 (link), 34 (link)].

Kobayashi H., Komatsu S., Ichikawa D., Kawaguchi T., Hirajima S., Miyamae M., Okajima W., Ohashi T., Kosuga T., Konishi H., Shiozaki A., Fujiwara H., Okamoto K., Tsuda H, & Otsuji E. (2015). Overexpression of denticleless E3 ubiquitin protein ligase homolog (DTL) is related to poor outcome in gastric carcinoma. Oncotarget, 6(34), 36615-36624.

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Transwell Migration and Invasion Assay

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Transwell migration and invasion assays were conducted in 24-well modified Boyden chambers (Transwell chambers, BD Transduction, Franklin Lakes, NJ). The upper surface of 6.4-mm-diameter filters with 8-μm pores was precoated with (invasion assay) or without (migration assay) Matrigel (BD Transduction). The miRNA mimic transfectants (5 × 10⁵ cells per well) were transferred into the upper chamber. Following 22 h of incubation, the migrated or invasive cells on the lower surface of the filters were fixed and stained with Diff-Quik stain (Sysmex, Kobe, Japan), and stained cell nuclei were counted directly in triplicate^{49 (link)}.

Nishibeppu K., Komatsu S., Imamura T., Kiuchi J., Kishimoto T., Arita T., Kosuga T., Konishi H., Kubota T., Shiozaki A., Fujiwara H., Okamoto K, & Otsuji E. (2020). Plasma microRNA profiles: identification of miR-1229-3p as a novel chemoresistant and prognostic biomarker in gastric cancer. Scientific Reports, 10, 3161.

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Invasion Assay for Cell Migration

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Invasion assays were undertaken using the BD BioCoat Matrigel Invasion Chamber (pore size 8 µm) (Becton Dickinson). Caki‐2 cells or ACHN cells transfected with the miRIDIAN hairpin or mimic for 72 hours were reseeded into the inserts of 24‐well plates (Caki‐2 cells, 2.0 × 10⁴ cells/well; ACHN, 1.5 × 10⁴ cells/well) in serum‐free media, with the base plate containing RPMI supplemented with 10% FBS. After 48 hours of incubation at 37℃, cells that had penetrated the Matrigel were fixed and stained using Diff‐Quik stain (Sysmex).

Wang C., Uemura M., Tomiyama E., Matsushita M., Koh Y., Nakano K., Hayashi Y., Ishizuya Y., Jingushi K., Kato T., Hatano K., Kawashima A., Ujike T., Nagahara A., Fujita K., Imamura R., Tsujikawa K, & Nonomura N. (2020). MicroRNA‐92b‐3p is a prognostic oncomiR that targets TSC1 in clear cell renal cell carcinoma. Cancer Science, 111(4), 1146-1155.

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Bronchoalveolar Lavage Fluid Analysis

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BALF was performed in the right lung through a tracheal cannula during exsanguination using 0.7 mL phosphate buffered saline (PBS) three times in each mouse. Total cell counts in the BALF were performed using a disposable hemocytometer (INCYTO, Seoul, Korea). A 0.2 mL aliquot of the cell suspension was centrifuged (Shandon Cytospin 4; Thermo, Waltham, MA, USA) at 800 rpm for 10 min onto a glass slide. The slides were stained with Diff-Quik stain (Sysmex, Kobe, Japan), and then washed, air-dried, and observed by a light microscope (BX51; Olympus, Tokyo, Japan). At least 200 cells included macrophages, neutrophils, and lymphocytes per sample were scored.

Kim H.Y., Kim M.S., Kim S.H., Joen D, & Lee K. (2018). Protective Effects of Nintedanib against Polyhexamethylene Guanidine Phosphate-Induced Lung Fibrosis in Mice. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(8), 1974.

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Transwell Migration and Invasion Assay

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Transwell migration and invasion assays were carried out in a 24‐well Transwell chamber using a cell culture insert with 8.0‐µm pores. The upper surface of the 6.4‐mm‐diameter filters with 8‐µm pores was precoated with (Corning BioCoat Matrigel Invasion Chamber; Corning) or without (Falcon Cell Culture Inserts; Corning) Matrigel. The siRNA transfectants (2.5 × 10⁵ cells per well) were seeded into the upper chamber with serum‐free medium. Complete growth medium was added to the lower well of each chamber. The transfectants were incubated for 22 h, then migrated or invasive cells on the lower surface of the filters were fixed and stained with Diff‐Quik stain (Sysmex). The stained cell nuclei were counted directly, in triplicate, as described elsewhere.³⁵, ³⁶

Takashima Y., Komatsu S., Ohashi T., Kiuchi J., Kamiya H., Shimizu H., Arita T., Konishi H., Shiozaki A., Kubota T., Okamoto K., Fujiwara H., Tsuda H, & Otsuji E. (2022). Overexpression of Tetraspanin31 contributes to malignant potential and poor outcomes in gastric cancer. Cancer Science, 113(6), 1984-1998.

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