In a separate experiment using adipocytes isolated from MRPVAT (for comparison), TA, and SMA/V PVAT, incubation with Mini202 (500 μM) was performed compared to a vehicle control so as to determine the generality of Mini202 uptake between adipocytes isolated from different vessels. Isolation and staining occurred exactly as described above. Cellular images were captured and analyzed on the Nexcelom Cellometer Vision® (Nexcelom Bioscience, Lawrence, MA) with the 450–302 cube. In these same cells, the viability of the adipocytes was assessed by uptake of the nucleic acid binding dyes acridine orange (AO) and propidium iodide (PI) (Cellometer AOPI #CS2–0106, Nexcelom Bioscience). AO is cell permeable (nuclear) and PI will only enter a cell with a compromised membrane, or an unhealthy/dying, dead or necrotic cells. Adipocytes from these experiments were imaged using the Cellometer Vision, 535–402 cube, 660–502 cube and data analyzed using Cellometer Vision software v.2.1.10.11.
Nexcelom cellometer vision
Manufactured by Revvity
The Nexcelom Cellometer Vision is a compact and automated cell counting and analysis system. It utilizes bright-field imaging and advanced software algorithms to provide cell count, viability, and size measurements for a variety of cell types. The Cellometer Vision is designed for researchers and laboratory professionals who require reliable and efficient cell analysis.
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Lab products found in correlation
2 protocols using nexcelom cellometer vision
1
Adipocyte Uptake of Mini202 Across PVAT
2
Yeast Expression Modulation Protocol
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Well-isolated single yeast colonies were picked from plates and incubated overnight in synthetic drop-out medium supplemented with 2% glucose at 30 °C. Twelve hours later, the 1-mL cell suspensions were diluted to the concentration 1 × 106 cells per mL—concentration estimated using a Nexcelom Cellometer Vision cell counter (Nexcelom Bioscience)—in fresh medium supplemented with 2% galactose. Triplicate cultures were grown for 48 h in galactose medium with the appropriate concentration of doxycycline (Acros Organics) for each condition to allow gene expression levels to stabilize (32 (link), 33 (link)). Cell density was measured (Nexcelom Cellometer Vision) and resuspended to a concentration 1 × 106 cells per mL every 12 or 24 h depending on the experimental condition to keep them in log growth phase.
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