To assess the effects of SLUG and TAL1-PP22 overexpression on GSC growth, 15,000 transduced Gb4 and Gb7 cells were dissociated, sorted for GFP positivity using a BD FACSAria III, and seeded as proliferating neurospheres in 24-well plates (n = 5 wells per conditions). After 5 days of growth, GSCs were dissociated with trypsin directly in the wells (0.5% final) and counted with an automated Z2 Coulter Cell Counter (Beckman Coulter, Villepinte, France). Cell growth measurements were repeated in 3 independent experiments for each cultures using the same protocol.
Z2 coulter cell counter
Manufactured by Beckman Coulter
Sourced in France
The Z2 Coulter Cell Counter is a laboratory instrument used for the automated counting and sizing of cells. It operates on the Coulter Principle, which measures changes in electrical impedance to determine the volume and number of cells passing through a small aperture. The Z2 Coulter Cell Counter provides accurate and reliable cell counts for a variety of cell types in research and clinical settings.
Automatically generated - may contain errors
Lab products found in correlation
Enzyme free cell dissociation buffer, by Thermo Fisher Scientific (2 mentions)
Facsaria 3, by BD (1 mentions)
Accuri c6 flow cytometer, by BD (1 mentions)
Alexa fluor 488 conjugated egf, by Thermo Fisher Scientific (1 mentions)
Filipin, by Merck Group (1 mentions)
Live cell imaging solution, by Thermo Fisher Scientific (1 mentions)
Lactose, by Merck Group (1 mentions)
4 protocols using z2 coulter cell counter
1
Analyzing GSC Growth from Transduced Cells
2
Investigating MKK5 Silencing in T. brucei Growth
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To investigate the impact of MKK5 silencing in the cell growth of procyclic forms of T.
brucei we evaluated the growth rate of cells silenced or not for MKK5. For this, two distinct strategies were employed. In the first one, the cell density was assessed every day until cells reached the stationary growth phase using a Neubauer chamber. In the second strategy, cell density was measured every two days using a Z2 Coulter cell counter (Beckman Coulter), followed by a dilution of the original culture to 1 x 10 6 cells/ml. In the second set-up, the cell growth was assessed for 10 days and cells were kept in exponential growth phase.
brucei we evaluated the growth rate of cells silenced or not for MKK5. For this, two distinct strategies were employed. In the first one, the cell density was assessed every day until cells reached the stationary growth phase using a Neubauer chamber. In the second strategy, cell density was measured every two days using a Z2 Coulter cell counter (Beckman Coulter), followed by a dilution of the original culture to 1 x 10 6 cells/ml. In the second set-up, the cell growth was assessed for 10 days and cells were kept in exponential growth phase.
3
Flow Cytometry Lectin Binding Assay
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Cells were incubated for 48 h with 1,3,4-O-Bu3ManNAc. The cells were washed with PBS, treated with enzyme free cell dissociation buffer (Life Technologies) until they detached from the culture plate, collected, and counted and cell numbers were normalized using the Beckman Z2 cell coulter counter. Cells were then washed twice in PBS. Cells were then incubated at room temperature for 120 min with 5.0 μg/mL of Flourescein-labeled RCA lectin (Vector Laboratories). Cells were washed three times in PBS and analyzed using flowcytometry with an Accuri C6 Flow cytometer. The cell population of interest was gated appropriately and 104 cells were used to determine mean fluorescence.
4
Exploring Cell Surface Glycosylation Dynamics
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Cells were incubated for 48 h with 1,3,4-O-Bu3ManNAc with serum starvation over the last 24 h. The cells then were washed with PBS, treated with enzyme free cell dissociation buffer (Life Technologies) until they detached from the culture plate, collected, and counted and cell numbers were normalized using the Beckman Z2 cell coulter counter. Cells were then washed twice in Live Cell Imaging Solution (Life Technologies) supplemented with 1.0% bovine serum albumin (BSA) and 20 mM glucose and treated with 0.5 μg/mL of filipin (Sigma Aldrich) or 100 mM of lactose (Carbosynth) for 60 min. Cells were then incubated at 37°C for 30 min with 2.0 μg/mL of Alexa Fluor 488-conjugated EGF (Life Technologies). Cells were washed three times, followed by acid washing for 5.0 min with 0.2 M glycine (pH 2.5), washed thrice and finally analyzed using flowcytometry with an Accuri C6 Flowcytometer. The cell population of interest was gated appropriately and 104 cells falling within the gated area were measured and used to determine the mean fluorescence of the cell population; the histograms for these experiments are shown in Figure S6 in the Supplemental Material .
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