Cell lines Panc1, Panc01.28, Panc06.05, Panc08.13, and Panc10.05 were a kind gift of Dr. Elizabeth Jaffe (Johns Hopkins Sidney Kimmel Comprehensive Cancer Center) and were grown as previously described [20 (link)]. Doubling time was determined by seeding 3×104 cells in triplicates in 6-well dishes and counting viable cells in triplicate for each well (trypan blue dye exclusion method using a T4 Cellometer (Nexcelom Bioscience, Lawrence, MA)) every 24 hours. The doubling time (DT) was calculated from the logarithmic portion of the growth curve using the formula DT = T × ln2 / ln(Xe/Xb) with T, incubation time in any units; Xb, cell number at beginning of incubation time; Xe, cell number at end of incubation time.
T4 cellometer
Manufactured by Revvity
Sourced in United States
The T4 Cellometer is a compact cell counter and viability analyzer designed for accurate and reliable cell counting. It utilizes advanced image-based technology to provide precise cell concentration and viability measurements for a variety of cell types.
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Lab products found in correlation
5 protocols using t4 cellometer
1
Determining Cell Line Doubling Time
2
Cell Seeding and Density Optimization for TF-1 Cells
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Cells were seeded in 384-well plates (ViewPlate-384 Black - PerkinElmer, ref. 6007460) using a MultiDrop combi (ThermoFisher Scientific), in 40 μL of cell media at 37 °C for 24 h. Optimal cell densities were determined as 5 × 103 for CTR and NF1KO_1 and 6 × 103 for NF1KO_2 TF-1 cell lines using T4 Cellometer (Nexcelom). Detailed procedures and data processing methods are provided in the Supplementary Material section.
3
Engineered HeLa Cell Seeding Protocol
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HeLa cells stably expressing Streptavidin-KDEL_ManII-SBP-EGFP (Boncompain et al., 2012 (link)) were counted using T4 Cellometer (Nexcelom) and seeded in 384-well plates (ViewPlate-384 Black Perkin Elmer, catalog number 6007460) at 2,000 cells/well using a Multidrop Combi (Thermo Fisher Scientific) in 40 μl of cell media. The screen was performed at same early cell passages in two replicate experiments.
4
Cultivation of Phaeodactylum tricornutum
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Phaeodactylum tricornutum Pt4 strain (UTEX 646) was grown in artificial sea water (Instant Ocean, Spectrum Brands, Blacksburg, VA, USA) supplemented with F/2 nutrients. Cultures were grown at 20°C under constant illumination (100 µmol m−2 sec−1, 4000 K White and 660 nm LED lighting) and agitated continuously at 70 rpm. Growth was monitored by OD750nm calibrated to cell density measured by an automated cell counter (Cellometer T4, Nexcelom, Lawrence, MA, USA). Lines were maintained on F/4 agar plates grown at 20°C under 50 µmol m−2 sec−1 (3500 K fluorescent tubes).
5
Trypan Blue Assay for AFB1 Cytotoxicity
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To measure cell viability after AFB1 exposure, we performed a trypan blue exclusion assay. Selected strains expressing CYP1A2 were inoculated in SC-URA until cultures reached an A600 ∼0.1-0.5, and then exposed to either 50 μM in AFB1 or DMSO (solvent) alone. After incubating for 3 hs, cells were washed twice in sterile phosphate buffered saline (PBS) and stained with trypan blue at a final concentration ∼10 μg/ml (Liesche et al. 2015 (link)). Cells were counted in a Nexcelom cellometer T4, according to the manufacturer’s instructions. A minimum of 104 cells were counted and all strains were tested at least twice. Statistical significance was determined by the Student’s t-test.
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