To enable exponential cell growth for the duration of the experiment, 1.5 − 2 × 106 cells in 5 mL fresh medium were seeded in 6-well plates and allowed to adhere overnight. Cells then were treated in triplicate with 4 different concentrations of gemcitabine and trabectedin, alone or combined, for 6 different time intervals of up to 120 h (Table 1 ). Equivalent volumes of water or DMSO were added as vehicle controls for the two drugs. At the appropriate times, triplicate wells of cells were washed twice with Dulbecco's phosphate buffered saline (Gibco) and suspended by incubating in 500 mL 1 × Trypsin EDTA (Cellgro) for 5 min. Cells were counted using a Coulter Counter model Z2 (Beckman Coulter, Hialeah, FL), and analyzed by flow cytometry.
Coulter counter model z2
Manufactured by Beckman Coulter
Sourced in United States
The Coulter Counter Model Z2 is a laboratory instrument designed for particle counting and sizing. It utilizes the Coulter principle to detect and measure the size distribution of particles suspended in an electrolyte solution. The core function of the Coulter Counter Model Z2 is to provide accurate and reproducible particle analysis data.
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Lab products found in correlation
4 protocols using coulter counter model z2
1
Exponential Cell Growth Optimization
2
Cell Volume Measurements under Osmotic Stress
3
Cell Volume Measurements under Osmotic Stress
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Cell volume measurements were performed by using a Coulter Counter Model Z2 (Beckman). Cells in suspension were diluted in PBS (isotonic control) or PBS with 100 mM NaCl, 200 mM sorbitol or 200 mM glucose (hypertonic conditions; + 200 mOsm/kg H2O) and incubated for the indicated times. Cells were analyzed according to manufacturer’s instructions. Cell sizes were also observed by differential interference contrast (DIC) microscopy.
4
Quantifying Steroid Hormone Levels in Cultured Cells
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The medium was collected from individual wells and frozen at −20 °C for subsequent determination of concentrations of estradiol and progesterone via RIA as previously described [24 (link),48 (link)]. A 24 h period of testosterone exposure to GC allows for a direct measure of functional aromatase activity [49 (link)]. The intra-assay coefficients of variation averaged 5.8% and 9.3% for the progesterone and estradiol RIA, respectively. The numbers of cells in the same wells in that medium was collected were determined as previously described [24 (link),48 (link),52 (link)] using a Coulter counter (model Z2; Beckman Coulter, Inc., Miami, FL, USA). The intra-assay coefficients of variation averaged 2.0 ± 0.8%.
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