Humidity control for Live-Cell imaging
Humidity control during live-cell, time-lapse experiments can be frustrating and lead to costly wasted time if you don’t have the right equipment and set up properly. It is important that the gas delivered to the chamber actually arrives at 100% humidity and the flow rate is appropriate for the size of the chamber. For single well time-lapse chambers, the airspace to culture medium ratio is low. Therefore, as a rule the volumetric gas exchange rate (assuming it is done properly) should be on the order of 1 to 5 volumes per hour to prevent pH or osmolarity shifts. The humidity in a closed cell chamber or dish reaches 100% within seconds of closure. Therefore, if you are applying a specialized gas mixture or 5% CO2 in air to the existing 100% RH airspace the inflow must be 100% RH or it will not take long to cause the pH and osmolarity of the media to change. The Bioptechs Micro-Gas Humidifier is an efficient, reliable, and convenient means to eliminate this obstacle for long-term imaging.
The Bioptechs Gas Humidifier warms water in a small containment vessel that the gas flows through creating a 100% RH atmosphere. The outflow of this gas is captured in the vessel, passed back through the heated base then close-coupled to the cell chamber thus preventing the RH from condensing before it reaches the cell chamber. If it is necessary to refill the water during an experiment, it has a Luer Slip refill cap so that water can be refilled with an ordinary syringe. It is safely powered by 12VDC and uses less than 2.5 watts. As a rule of thumb an unheated bubbler will only raise the RH around 10 to 15% above the inflow RH. However, a properly heated bubbler of the appropriate geometry will raise the inflow RH to 100%. It should also be appreciated that although you can produce 100% RH in a bottle or other device external to the cell containment structure it is necessary to prevent the air/gas flowing through the humidification device from condensing on its way to the cell chamber. Bioptechs addresses this problem by directly coupling the humidifier to the cell chamber thus providing a convenient, trouble free, and reliable solution. The Bioptechs Micro-Gas Humidifier along with a simple combination of a demand regulator and low flow peristaltic pump enables anyone to easily and cost effectively establish the proper gas and RH conditions for live cell microscopy.How it works:
This technique is ideal for long-term micro-observation of adherent cell culture, tissue slices in media, and suspended cells in CO2 dependent media on an inverted microscope or upright microscope when combined with the appropriate accessories.
Bioptechs Humidifier Components
|Leur taper fill port cap, humidifier top with gas inlet port, humidifier glass, humidifier bottom with gas output port and power adapter port|