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diff-quick fixative  (Dade Behring)

 
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    Dade Behring diff-quick fixative
    Diff Quick Fixative, supplied by Dade Behring, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/diff+quick+fixative/diff+quick+fixative+solutions/pmc07426628-70-30-32
    Average 90 stars, based on 1 article reviews
    diff-quick fixative - by Bioz Stars, 2026-10
    90/100 stars

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    Related Articles

    Incubation:

    Article Title: PI3K/AKT/mTOR and sonic hedgehog pathways cooperate together to inhibit human pancreatic cancer stem cell characteristics and tumor growth
    Article Snippet: After 24 h of incubation, cells were fixed with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring, Newark, DE, USA).

    Article Title: Anti-angiogenesis triggers exosomes release from endothelial cells to promote tumor vasculogenesis
    Article Snippet: After 6 h (HUVEC angiogenesis) or 8 h (VM by HCC cells) incubation at 37°C in a 5% CO 2 /95% air incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, USA) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: Hematology, Serum Chemistry, and Early Hematologic Changes in Free-Ranging South American Fur Seals ( Arctocephalus australis ) at Guafo Island, Chilean Patagonia.
    Article Snippet: The establishment of clinical pathology baseline data is critical to evaluate temporal and spatial changes in marine mammal groups.. Despite increased availability of studies on hematology and biochemistry of marine mammals, reference ranges are lacking for many populations, especially among fur seal species.. During the austral summers of 2014 and 2015, we evaluated basic hematologic and biochemical parameters in clinically healthy, physically restrained South American fur seal (Arctocephalus australis) lactating females and 2-mo-old pups.

    Article Title: Anti-angiogenesis triggers exosomes release from endothelial cells to promote tumor vasculogenesis.
    Article Snippet: After 6 h (HUVEC angiogenesis) or 8 h (VM by HCC cells) incubation at 37°C in a 5% CO2/95% air incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, USA) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: MiR-9 Promotes Angiogenesis via Targeting on Sphingosine-1- Phosphate Receptor 1
    Article Snippet: After 6 h incubation with or without 50 ng/mL hVEGF at 37°C with 5% CO in a humidified incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, United States) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: NPV-LDE-225 (Erismodegib) inhibits epithelial mesenchymal transition and self-renewal of glioblastoma initiating cells by regulating miR-21, miR-128, and miR-200
    Article Snippet: Cells were fixed after 24 h of incubation with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring).

    Article Title: MiR-9 Promotes Angiogenesis via Targeting on Sphingosine-1- Phosphate Receptor 1.
    Article Snippet: After 6 h incubation with or without 50 ng/mL hVEGF at 37◦C with 5% CO2 in a humidified incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, United States) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: NVP-LDE-225 (Erismodegib) inhibits epithelial-mesenchymal transition and human prostate cancer stem cell growth in NOD/SCID IL2Rγ null mice by regulating Bmi-1 and microRNA-128.
    Article Snippet: Cells were fixed after 24 h of incubation with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring, Newark, DE, USA).

    Diff-Quik:

    Article Title: PI3K/AKT/mTOR and sonic hedgehog pathways cooperate together to inhibit human pancreatic cancer stem cell characteristics and tumor growth
    Article Snippet: After 24 h of incubation, cells were fixed with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring, Newark, DE, USA).

    Article Title: Anti-angiogenesis triggers exosomes release from endothelial cells to promote tumor vasculogenesis
    Article Snippet: After 6 h (HUVEC angiogenesis) or 8 h (VM by HCC cells) incubation at 37°C in a 5% CO 2 /95% air incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, USA) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: Hematology, Serum Chemistry, and Early Hematologic Changes in Free-Ranging South American Fur Seals ( Arctocephalus australis ) at Guafo Island, Chilean Patagonia.
    Article Snippet: The establishment of clinical pathology baseline data is critical to evaluate temporal and spatial changes in marine mammal groups.. Despite increased availability of studies on hematology and biochemistry of marine mammals, reference ranges are lacking for many populations, especially among fur seal species.. During the austral summers of 2014 and 2015, we evaluated basic hematologic and biochemical parameters in clinically healthy, physically restrained South American fur seal (Arctocephalus australis) lactating females and 2-mo-old pups.

    Article Title: Anti-angiogenesis triggers exosomes release from endothelial cells to promote tumor vasculogenesis.
    Article Snippet: After 6 h (HUVEC angiogenesis) or 8 h (VM by HCC cells) incubation at 37°C in a 5% CO2/95% air incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, USA) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: MiR-9 Promotes Angiogenesis via Targeting on Sphingosine-1- Phosphate Receptor 1
    Article Snippet: After 6 h incubation with or without 50 ng/mL hVEGF at 37°C with 5% CO in a humidified incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, United States) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: NPV-LDE-225 (Erismodegib) inhibits epithelial mesenchymal transition and self-renewal of glioblastoma initiating cells by regulating miR-21, miR-128, and miR-200
    Article Snippet: Cells were fixed after 24 h of incubation with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring).

    Article Title: MiR-9 Promotes Angiogenesis via Targeting on Sphingosine-1- Phosphate Receptor 1.
    Article Snippet: After 6 h incubation with or without 50 ng/mL hVEGF at 37◦C with 5% CO2 in a humidified incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, United States) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: NVP-LDE-225 (Erismodegib) inhibits epithelial-mesenchymal transition and human prostate cancer stem cell growth in NOD/SCID IL2Rγ null mice by regulating Bmi-1 and microRNA-128.
    Article Snippet: Cells were fixed after 24 h of incubation with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring, Newark, DE, USA).

    Staining:

    Article Title: PI3K/AKT/mTOR and sonic hedgehog pathways cooperate together to inhibit human pancreatic cancer stem cell characteristics and tumor growth
    Article Snippet: After 24 h of incubation, cells were fixed with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring, Newark, DE, USA).

    Article Title: Anti-angiogenesis triggers exosomes release from endothelial cells to promote tumor vasculogenesis
    Article Snippet: After 6 h (HUVEC angiogenesis) or 8 h (VM by HCC cells) incubation at 37°C in a 5% CO 2 /95% air incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, USA) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: Hematology, Serum Chemistry, and Early Hematologic Changes in Free-Ranging South American Fur Seals ( Arctocephalus australis ) at Guafo Island, Chilean Patagonia.
    Article Snippet: The establishment of clinical pathology baseline data is critical to evaluate temporal and spatial changes in marine mammal groups.. Despite increased availability of studies on hematology and biochemistry of marine mammals, reference ranges are lacking for many populations, especially among fur seal species.. During the austral summers of 2014 and 2015, we evaluated basic hematologic and biochemical parameters in clinically healthy, physically restrained South American fur seal (Arctocephalus australis) lactating females and 2-mo-old pups.

    Article Title: Anti-angiogenesis triggers exosomes release from endothelial cells to promote tumor vasculogenesis.
    Article Snippet: After 6 h (HUVEC angiogenesis) or 8 h (VM by HCC cells) incubation at 37°C in a 5% CO2/95% air incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, USA) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: MiR-9 Promotes Angiogenesis via Targeting on Sphingosine-1- Phosphate Receptor 1
    Article Snippet: After 6 h incubation with or without 50 ng/mL hVEGF at 37°C with 5% CO in a humidified incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, United States) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: NPV-LDE-225 (Erismodegib) inhibits epithelial mesenchymal transition and self-renewal of glioblastoma initiating cells by regulating miR-21, miR-128, and miR-200
    Article Snippet: Cells were fixed after 24 h of incubation with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring).

    Article Title: MiR-9 Promotes Angiogenesis via Targeting on Sphingosine-1- Phosphate Receptor 1.
    Article Snippet: After 6 h incubation with or without 50 ng/mL hVEGF at 37◦C with 5% CO2 in a humidified incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, United States) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: NVP-LDE-225 (Erismodegib) inhibits epithelial-mesenchymal transition and human prostate cancer stem cell growth in NOD/SCID IL2Rγ null mice by regulating Bmi-1 and microRNA-128.
    Article Snippet: Cells were fixed after 24 h of incubation with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring, Newark, DE, USA).

    Microscopy:

    Article Title: PI3K/AKT/mTOR and sonic hedgehog pathways cooperate together to inhibit human pancreatic cancer stem cell characteristics and tumor growth
    Article Snippet: After 24 h of incubation, cells were fixed with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring, Newark, DE, USA).

    Article Title: Anti-angiogenesis triggers exosomes release from endothelial cells to promote tumor vasculogenesis
    Article Snippet: After 6 h (HUVEC angiogenesis) or 8 h (VM by HCC cells) incubation at 37°C in a 5% CO 2 /95% air incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, USA) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: Hematology, Serum Chemistry, and Early Hematologic Changes in Free-Ranging South American Fur Seals ( Arctocephalus australis ) at Guafo Island, Chilean Patagonia.
    Article Snippet: The establishment of clinical pathology baseline data is critical to evaluate temporal and spatial changes in marine mammal groups.. Despite increased availability of studies on hematology and biochemistry of marine mammals, reference ranges are lacking for many populations, especially among fur seal species.. During the austral summers of 2014 and 2015, we evaluated basic hematologic and biochemical parameters in clinically healthy, physically restrained South American fur seal (Arctocephalus australis) lactating females and 2-mo-old pups.

    Article Title: Anti-angiogenesis triggers exosomes release from endothelial cells to promote tumor vasculogenesis.
    Article Snippet: After 6 h (HUVEC angiogenesis) or 8 h (VM by HCC cells) incubation at 37°C in a 5% CO2/95% air incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, USA) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: MiR-9 Promotes Angiogenesis via Targeting on Sphingosine-1- Phosphate Receptor 1
    Article Snippet: After 6 h incubation with or without 50 ng/mL hVEGF at 37°C with 5% CO in a humidified incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, United States) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: NPV-LDE-225 (Erismodegib) inhibits epithelial mesenchymal transition and self-renewal of glioblastoma initiating cells by regulating miR-21, miR-128, and miR-200
    Article Snippet: Cells were fixed after 24 h of incubation with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring).

    Article Title: MiR-9 Promotes Angiogenesis via Targeting on Sphingosine-1- Phosphate Receptor 1.
    Article Snippet: After 6 h incubation with or without 50 ng/mL hVEGF at 37◦C with 5% CO2 in a humidified incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, United States) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: NVP-LDE-225 (Erismodegib) inhibits epithelial-mesenchymal transition and human prostate cancer stem cell growth in NOD/SCID IL2Rγ null mice by regulating Bmi-1 and microRNA-128.
    Article Snippet: Cells were fixed after 24 h of incubation with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring, Newark, DE, USA).

    Migration:

    Article Title: PI3K/AKT/mTOR and sonic hedgehog pathways cooperate together to inhibit human pancreatic cancer stem cell characteristics and tumor growth
    Article Snippet: After 24 h of incubation, cells were fixed with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring, Newark, DE, USA).

    Article Title: Anti-angiogenesis triggers exosomes release from endothelial cells to promote tumor vasculogenesis
    Article Snippet: After 6 h (HUVEC angiogenesis) or 8 h (VM by HCC cells) incubation at 37°C in a 5% CO 2 /95% air incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, USA) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: Hematology, Serum Chemistry, and Early Hematologic Changes in Free-Ranging South American Fur Seals ( Arctocephalus australis ) at Guafo Island, Chilean Patagonia.
    Article Snippet: The establishment of clinical pathology baseline data is critical to evaluate temporal and spatial changes in marine mammal groups.. Despite increased availability of studies on hematology and biochemistry of marine mammals, reference ranges are lacking for many populations, especially among fur seal species.. During the austral summers of 2014 and 2015, we evaluated basic hematologic and biochemical parameters in clinically healthy, physically restrained South American fur seal (Arctocephalus australis) lactating females and 2-mo-old pups.

    Article Title: Anti-angiogenesis triggers exosomes release from endothelial cells to promote tumor vasculogenesis.
    Article Snippet: After 6 h (HUVEC angiogenesis) or 8 h (VM by HCC cells) incubation at 37°C in a 5% CO2/95% air incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, USA) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: MiR-9 Promotes Angiogenesis via Targeting on Sphingosine-1- Phosphate Receptor 1
    Article Snippet: After 6 h incubation with or without 50 ng/mL hVEGF at 37°C with 5% CO in a humidified incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, United States) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: NPV-LDE-225 (Erismodegib) inhibits epithelial mesenchymal transition and self-renewal of glioblastoma initiating cells by regulating miR-21, miR-128, and miR-200
    Article Snippet: Cells were fixed after 24 h of incubation with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring).

    Article Title: MiR-9 Promotes Angiogenesis via Targeting on Sphingosine-1- Phosphate Receptor 1.
    Article Snippet: After 6 h incubation with or without 50 ng/mL hVEGF at 37◦C with 5% CO2 in a humidified incubator, the medium was gently aspirated from each well and incubated with Diff-Quick fixative (Dade Behring, Deerfield, IL, United States) for 30 s and subsequently stained with solution II for 2 min. Tube structures were observed and imaged by microscopy.

    Article Title: NVP-LDE-225 (Erismodegib) inhibits epithelial-mesenchymal transition and human prostate cancer stem cell growth in NOD/SCID IL2Rγ null mice by regulating Bmi-1 and microRNA-128.
    Article Snippet: Cells were fixed after 24 h of incubation with methanol and stained with Diff-Quick Fixative Solutions (Dade Behring, Newark, DE, USA).



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    Image Search Results


    Glycolysis stress test profile of L. donovani -infected neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C and 5% CO 2 . Uninfected cells served as control. The infection rate was determined by Giemsa staining of cytocentrifuged samples. Subsequently, free, non-ingested parasites were removed by washing. After 6 h post infection the glycolysis stress test profile was determined by using the Seahorse extracellular flux analyzer (A) . Extracellular acidification rate (ECAR) measurements following the sequential injection of 5 mM glucose, 1 μM oligomycin, and 10 mM 2-DG (dotted lines indicate injection time) were used to calculate key parameters of glycolytic function. Glycolysis (B) was calculated by subtraction of 2-DG-mediated ECAR from glucose-mediated ECAR. Glycolytic capacity (C) was calculated by subtraction of 2-DG-mediated ECAR from oligomycin-mediated ECAR. Bar graphs show mean ± SD ( n = 8), ** p ≤ 0.01.

    Journal: Frontiers in Immunology

    Article Title: Enhanced Glycolysis Is Required for Antileishmanial Functions of Neutrophils Upon Infection With Leishmania donovani

    doi: 10.3389/fimmu.2021.632512

    Figure Lengend Snippet: Glycolysis stress test profile of L. donovani -infected neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C and 5% CO 2 . Uninfected cells served as control. The infection rate was determined by Giemsa staining of cytocentrifuged samples. Subsequently, free, non-ingested parasites were removed by washing. After 6 h post infection the glycolysis stress test profile was determined by using the Seahorse extracellular flux analyzer (A) . Extracellular acidification rate (ECAR) measurements following the sequential injection of 5 mM glucose, 1 μM oligomycin, and 10 mM 2-DG (dotted lines indicate injection time) were used to calculate key parameters of glycolytic function. Glycolysis (B) was calculated by subtraction of 2-DG-mediated ECAR from glucose-mediated ECAR. Glycolytic capacity (C) was calculated by subtraction of 2-DG-mediated ECAR from oligomycin-mediated ECAR. Bar graphs show mean ± SD ( n = 8), ** p ≤ 0.01.

    Article Snippet: The preparations contained ≥99% granulocytes, of which >95% were neutrophils and 1–4 % were eosinophils, as determined by Giemsa staining (Diff Quick Fix, Medion Diagnostics, Berlin, Germany) of cytocentrifuged (Shandon) samples.

    Techniques: Infection, Control, Staining, Injection

    Mitochondrial stress test profile of L. donovani -infected neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C and 5% CO 2 . Uninfected cells served as control. Successful infection was determined by Giemsa staining of cytocentrifuged samples. Subsequently, free, non-ingested parasites were removed by washing. After 6 h post infection the mitochondrial stress test profile was determined by using the Seahorse extracellular flux analyzer (A) . The measurement of basal oxygen consumption rate (OCR) was followed by sequential injections of 1 μM oligomycin, 1.5 μM FCCP, and 1 μM rotenone/antimycin A (dotted lines indicate injection time). OCR measurements were used to calculate key parameters of mitochondrial function. Non-mitochondrial respiration (B) was calculated as OCR after rotenone/antimycin A injection. Basal respiration (C) was calculated by subtraction of rotenone/antimycin A-mediated OCR from basal OCR. Maximal respiration (D) was calculated by subtraction of rotenone/antimycin A-mediated OCR from FCCP-mediated OCR. Proton leak (E) was calculated by subtraction of non-mitochondrial respiration from oligomycin-mediated OCR. ATP production (F) was calculated by subtraction of oligomycin-mediated OCR from basal OCR. Bar graphs show mean ± SD ( n = 3), ** p ≤ 0.01, ns, not significant.

    Journal: Frontiers in Immunology

    Article Title: Enhanced Glycolysis Is Required for Antileishmanial Functions of Neutrophils Upon Infection With Leishmania donovani

    doi: 10.3389/fimmu.2021.632512

    Figure Lengend Snippet: Mitochondrial stress test profile of L. donovani -infected neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C and 5% CO 2 . Uninfected cells served as control. Successful infection was determined by Giemsa staining of cytocentrifuged samples. Subsequently, free, non-ingested parasites were removed by washing. After 6 h post infection the mitochondrial stress test profile was determined by using the Seahorse extracellular flux analyzer (A) . The measurement of basal oxygen consumption rate (OCR) was followed by sequential injections of 1 μM oligomycin, 1.5 μM FCCP, and 1 μM rotenone/antimycin A (dotted lines indicate injection time). OCR measurements were used to calculate key parameters of mitochondrial function. Non-mitochondrial respiration (B) was calculated as OCR after rotenone/antimycin A injection. Basal respiration (C) was calculated by subtraction of rotenone/antimycin A-mediated OCR from basal OCR. Maximal respiration (D) was calculated by subtraction of rotenone/antimycin A-mediated OCR from FCCP-mediated OCR. Proton leak (E) was calculated by subtraction of non-mitochondrial respiration from oligomycin-mediated OCR. ATP production (F) was calculated by subtraction of oligomycin-mediated OCR from basal OCR. Bar graphs show mean ± SD ( n = 3), ** p ≤ 0.01, ns, not significant.

    Article Snippet: The preparations contained ≥99% granulocytes, of which >95% were neutrophils and 1–4 % were eosinophils, as determined by Giemsa staining (Diff Quick Fix, Medion Diagnostics, Berlin, Germany) of cytocentrifuged (Shandon) samples.

    Techniques: Infection, Control, Staining, Injection

    2-NBDG uptake of L. donovani -infected neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C and 5 % CO 2 . Uninfected cells served as control. The infection rate was determined by Giemsa staining of cytocentrifuged samples. Subsequently, free, non-ingested parasites were removed by washing. After 6 h post infection the fluorescent glucose analog 2-NBDG was added to the cells in glucose-free medium for the last 10 min of incubation time and 2-NBDG uptake was analyzed by flow cytometry. The bar diagram shows the autofluorescence corrected mean fluorescence intensity (MFI) of 2-NBDG uptake ± SD ( n = 3), *** p ≤ 0.001.

    Journal: Frontiers in Immunology

    Article Title: Enhanced Glycolysis Is Required for Antileishmanial Functions of Neutrophils Upon Infection With Leishmania donovani

    doi: 10.3389/fimmu.2021.632512

    Figure Lengend Snippet: 2-NBDG uptake of L. donovani -infected neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C and 5 % CO 2 . Uninfected cells served as control. The infection rate was determined by Giemsa staining of cytocentrifuged samples. Subsequently, free, non-ingested parasites were removed by washing. After 6 h post infection the fluorescent glucose analog 2-NBDG was added to the cells in glucose-free medium for the last 10 min of incubation time and 2-NBDG uptake was analyzed by flow cytometry. The bar diagram shows the autofluorescence corrected mean fluorescence intensity (MFI) of 2-NBDG uptake ± SD ( n = 3), *** p ≤ 0.001.

    Article Snippet: The preparations contained ≥99% granulocytes, of which >95% were neutrophils and 1–4 % were eosinophils, as determined by Giemsa staining (Diff Quick Fix, Medion Diagnostics, Berlin, Germany) of cytocentrifuged (Shandon) samples.

    Techniques: Infection, Control, Staining, Incubation, Flow Cytometry, Fluorescence

    Lactate secretion and pyruvate content of L. donovani -infected neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C and 5% CO 2 . Uninfected cells served as control. The infection rate was determined by Giemsa staining of cytocentrifuged samples. Subsequently, free, non-ingested parasites were removed by washing. Whole cell lysates were generated after 6 h post infection by boiling cells at 90°C for 5 min. Cell-free supernatants were collected after 6 h post infection. Lactate was detected in culture supernatants by using a lactate assay kit. Pyruvate was detected in whole cell lysates by using a pyruvate assay kit. Bar diagrams show mean concentration of lactate (A) after subtraction of the medium blanks and pyruvate (B) calculated by interpolation from standard curve ± SD ( n = 5), * p ≤ 0.05, **** p ≤ 0.0001.

    Journal: Frontiers in Immunology

    Article Title: Enhanced Glycolysis Is Required for Antileishmanial Functions of Neutrophils Upon Infection With Leishmania donovani

    doi: 10.3389/fimmu.2021.632512

    Figure Lengend Snippet: Lactate secretion and pyruvate content of L. donovani -infected neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C and 5% CO 2 . Uninfected cells served as control. The infection rate was determined by Giemsa staining of cytocentrifuged samples. Subsequently, free, non-ingested parasites were removed by washing. Whole cell lysates were generated after 6 h post infection by boiling cells at 90°C for 5 min. Cell-free supernatants were collected after 6 h post infection. Lactate was detected in culture supernatants by using a lactate assay kit. Pyruvate was detected in whole cell lysates by using a pyruvate assay kit. Bar diagrams show mean concentration of lactate (A) after subtraction of the medium blanks and pyruvate (B) calculated by interpolation from standard curve ± SD ( n = 5), * p ≤ 0.05, **** p ≤ 0.0001.

    Article Snippet: The preparations contained ≥99% granulocytes, of which >95% were neutrophils and 1–4 % were eosinophils, as determined by Giemsa staining (Diff Quick Fix, Medion Diagnostics, Berlin, Germany) of cytocentrifuged (Shandon) samples.

    Techniques: Infection, Control, Staining, Generated, Lactate Assay, Pyruvate Assay, Concentration Assay

    ATP concentration in L. donovani -infected neutrophils as response to metabolic inhibitors. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C and 5% CO 2 . Uninfected cells served as control. The infection rate was determined by Giemsa staining of cytocentrifuged samples. Subsequently, free, non-ingested parasites were removed by washing. After 6 h post infection cells were treated with 10 mM 2-DG for 3 h at 37°C and 5% CO 2 . PBS served as solvent control. Whole cell lysates were prepared and the ATP concentration was determined by using the ATP determination kit. Bar diagrams show the mean ATP concentration ± SD ( n = 3) in uninfected (A) and L. donovani -infected neutrophils (B) , * p ≤ 0.05, *** p ≤ 0.001, **** p ≤ 0.0001.

    Journal: Frontiers in Immunology

    Article Title: Enhanced Glycolysis Is Required for Antileishmanial Functions of Neutrophils Upon Infection With Leishmania donovani

    doi: 10.3389/fimmu.2021.632512

    Figure Lengend Snippet: ATP concentration in L. donovani -infected neutrophils as response to metabolic inhibitors. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C and 5% CO 2 . Uninfected cells served as control. The infection rate was determined by Giemsa staining of cytocentrifuged samples. Subsequently, free, non-ingested parasites were removed by washing. After 6 h post infection cells were treated with 10 mM 2-DG for 3 h at 37°C and 5% CO 2 . PBS served as solvent control. Whole cell lysates were prepared and the ATP concentration was determined by using the ATP determination kit. Bar diagrams show the mean ATP concentration ± SD ( n = 3) in uninfected (A) and L. donovani -infected neutrophils (B) , * p ≤ 0.05, *** p ≤ 0.001, **** p ≤ 0.0001.

    Article Snippet: The preparations contained ≥99% granulocytes, of which >95% were neutrophils and 1–4 % were eosinophils, as determined by Giemsa staining (Diff Quick Fix, Medion Diagnostics, Berlin, Germany) of cytocentrifuged (Shandon) samples.

    Techniques: Concentration Assay, Infection, Control, Staining, Solvent

    Survival of L. donovani promastigotes in 2-DG-treated neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C, 5% CO 2 . The infection rate was determined by Giemsa staining of cytocentrifuged samples. After removing the free, non-ingested parasites the infected cells were treated with PBS, 5 mM, 50 mM, or 100 mM 2-DG. Survival of parasites was assessed after 24 h post infection by using the limiting dilution assay. The bar diagram shows the mean survival rates (%) normalized to PBS-treated control cells ± SD ( n = 3), **** p ≤ 0.0001.

    Journal: Frontiers in Immunology

    Article Title: Enhanced Glycolysis Is Required for Antileishmanial Functions of Neutrophils Upon Infection With Leishmania donovani

    doi: 10.3389/fimmu.2021.632512

    Figure Lengend Snippet: Survival of L. donovani promastigotes in 2-DG-treated neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C, 5% CO 2 . The infection rate was determined by Giemsa staining of cytocentrifuged samples. After removing the free, non-ingested parasites the infected cells were treated with PBS, 5 mM, 50 mM, or 100 mM 2-DG. Survival of parasites was assessed after 24 h post infection by using the limiting dilution assay. The bar diagram shows the mean survival rates (%) normalized to PBS-treated control cells ± SD ( n = 3), **** p ≤ 0.0001.

    Article Snippet: The preparations contained ≥99% granulocytes, of which >95% were neutrophils and 1–4 % were eosinophils, as determined by Giemsa staining (Diff Quick Fix, Medion Diagnostics, Berlin, Germany) of cytocentrifuged (Shandon) samples.

    Techniques: Infection, Staining, Limiting Dilution Assay, Control

    ROS production of 2-DG-treated L. donovani -infected neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C, 5% CO 2 . The infection rate was determined by Giemsa staining of cytocentrifuged samples. After removing the free, non-ingested parasites the infected and uninfected cells were treated with PBS, 5 mM, 50 mM, or 100 mM 2-DG. After 24 h post infection the MPO-derived ROS production was measured for 1 h at 37°C and 5% CO 2 after the stimulation with 20 nM PMA by using the luminol-based chemiluminescence assay. A representative curve of luminol chemiluminescence is shown in panel (A) . The bar diagram (B) shows the mean area under the curve (AUC) values ± SD ( n = 3), * p ≤ 0.05.

    Journal: Frontiers in Immunology

    Article Title: Enhanced Glycolysis Is Required for Antileishmanial Functions of Neutrophils Upon Infection With Leishmania donovani

    doi: 10.3389/fimmu.2021.632512

    Figure Lengend Snippet: ROS production of 2-DG-treated L. donovani -infected neutrophils. Primary human neutrophils were infected with L. donovani promastigotes (ratio 1:10) for 3 h at 37°C, 5% CO 2 . The infection rate was determined by Giemsa staining of cytocentrifuged samples. After removing the free, non-ingested parasites the infected and uninfected cells were treated with PBS, 5 mM, 50 mM, or 100 mM 2-DG. After 24 h post infection the MPO-derived ROS production was measured for 1 h at 37°C and 5% CO 2 after the stimulation with 20 nM PMA by using the luminol-based chemiluminescence assay. A representative curve of luminol chemiluminescence is shown in panel (A) . The bar diagram (B) shows the mean area under the curve (AUC) values ± SD ( n = 3), * p ≤ 0.05.

    Article Snippet: The preparations contained ≥99% granulocytes, of which >95% were neutrophils and 1–4 % were eosinophils, as determined by Giemsa staining (Diff Quick Fix, Medion Diagnostics, Berlin, Germany) of cytocentrifuged (Shandon) samples.

    Techniques: Infection, Staining, Derivative Assay, Chemiluminescence Immunoassay