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Beckman liquid scintillation counter ls 6500

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The Beckman Liquid Scintillation Counter LS-6500 is a laboratory instrument designed to detect and measure radioactive samples. It utilizes liquid scintillation technology to quantify the amount of radioactivity present in a sample.

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Erythrocyte depletion kit, by Miltenyi Biotec (2 mentions) Liquid scintillation cocktail, by Thermo Fisher Scientific (1 mentions)

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3 protocols using beckman liquid scintillation counter ls 6500

1

NK Cell-Mediated Cytotoxicity Assay

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U87, Gli36, U251 and GBM30 cells were infected with OV-Q1, OV-CDH1 or OV-IL2RA-CDH1 at a MOI of 5. Eight hours after infection the infected cells were used as target cells. Effector cells were primary human NK cells isolated from leukopaks of health donors using an NK cell isolation kit (MACSxpress® Miltenyi Biotec, San Diego, CA) and an erythrocyte depletion kit (Miltenyi Biotec). Target cells were labeled with Chromium-51 (51Cr) for 1 hour, and then co-cultured with bulk human primary NK cells or sorted KLRG1+ and KLRG1− NK cells at different effector:target ratios at 37℃ for 4 hours. Release of 51Cr was measured with a Beckman Liquid Scintillation Counter LS-6500 (Beckman Coulter, Fullerton, CA). Target cells incubated in complete media or 1% SDS media were used for spontaneous or maximal 51Cr release control, respectively. The cell lysis percentages were calculated using the standard formula: 100 × (cpm experimental release – cpm spontaneous release) / (cpm maximal release – cpm spontaneous release). The assays were performed in at least three technical replicates with NK cells from different donors.
Xu B., Ma R., Russell L., Yoo J.Y., Han J., Cui H., Yi P., Zhang J., Nakashima H., Dai H., Chiocca E.A., Kaur B., Caligiuri M.A, & Yu J. (2018). An oncolytic herpes virus expressing E-cadherin resists NK cell clearance and improves viral spread and glioblastoma virotherapy. Nature biotechnology.
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2

NK Cell-Mediated Cytotoxicity Assay

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U87, Gli36, U251 and GBM30 cells were infected with OV-Q1, OV-CDH1 or OV-IL2RA-CDH1 at a MOI of 5. Eight hours after infection the infected cells were used as target cells. Effector cells were primary human NK cells isolated from leukopaks of health donors using an NK cell isolation kit (MACSxpress® Miltenyi Biotec, San Diego, CA) and an erythrocyte depletion kit (Miltenyi Biotec). Target cells were labeled with Chromium-51 (51Cr) for 1 hour, and then co-cultured with bulk human primary NK cells or sorted KLRG1+ and KLRG1− NK cells at different effector:target ratios at 37℃ for 4 hours. Release of 51Cr was measured with a Beckman Liquid Scintillation Counter LS-6500 (Beckman Coulter, Fullerton, CA). Target cells incubated in complete media or 1% SDS media were used for spontaneous or maximal 51Cr release control, respectively. The cell lysis percentages were calculated using the standard formula: 100 × (cpm experimental release – cpm spontaneous release) / (cpm maximal release – cpm spontaneous release). The assays were performed in at least three technical replicates with NK cells from different donors.
Xu B., Ma R., Russell L., Yoo J.Y., Han J., Cui H., Yi P., Zhang J., Nakashima H., Dai H., Chiocca E.A., Kaur B., Caligiuri M.A, & Yu J. (2018). An oncolytic herpes virus expressing E-cadherin resists NK cell clearance and improves viral spread and glioblastoma virotherapy. Nature biotechnology.
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3

Chromium Release Cytotoxicity Assay

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A standard 51Cr release assay was performed as described previously (15 (link)). Briefly, target cells were labeled with 51Cr and co-cultured with the NK-92 cell line or freshly isolated primary NK cells pretreated with or without TGF-β1 at 20 ng/ml for 24 h at various effector:target (E:T) ratios in the wells of 96-well V-bottom plates at 37°C for 8 h. GB cell lines or patient-derived GB stem-like cells as target cells were pre-incubated with or without oHSV for 30 min at 37°C (multiplicity of infection, MOI=3). Supernatants were harvested and transferred into scintillation vials containing a liquid scintillation cocktail (Fisher Scientific, Waltham, MA), and the release of 51Cr was measured on Beckman Liquid Scintillation Counter LS-6500 (Beckman Coulter, Fullerton, CA). Target cells incubated in complete medium or 1% SDS were used to determine spontaneous or maximal 51Cr release, respectively. Percentage of specific cell lysis was calculated using the standard formula: 100 × (cpm experimental release – cpm spontaneous release) / (cpm maximal release – cpm spontaneous release).
Han J., Chen X., Chu J., Xu B., Meisen W.H., Chen L., Zhang L., Zhang J., He X., Wang Q.E., Chiocca E.A., Kaur B., Caligiuri M.A, & Yu J. (2015). TGF-β treatment enhances glioblastoma virotherapy by inhibiting the innate immune response. Cancer research, 75(24), 5273-5282.
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