In Vivo Scientific CO2 Systems

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CO2 Systems

We at In Vivo Scientific realize that many investigators are new to live cell microscopy, which presents different challenges not present in fixed-cell/traditional microscopy. To this end, we offer peripheral accessories ideally suited for live cell microscopy. These, when combined with our microscope incubator, provide a complete solution to most live cell imaging requirements.

In Vivo Scientific now offers three CO2 environment systems for live-cell microscopy for use in conjunction with our live-cell heated microscope incubator.
1) IVS-3000: Non-adjustable CO2 environment.
2) IVS-3001: Adjustable CO2 environment, without sample feedback control.
3) IVS-3003: Adjustable CO2 environment, with sample feedback control.

IVS-3000:
This system utilizes pre-mixed CO2 gas cylinders that are readily available from any local gas supplier (e.g. 5% CO2, 95% ambient air). Our design is similar in some ways to the CO2 system that leading competitors use, but with some distinct advantages. Dry, pre-mixed 5% CO2 exits the gas cylinder, passes through a pressure regulator set to 100 mm Hg. It then passes through a flow regulator that can be set to the user's preferences (usually 30-50 mL/minute). This low-flow gas then enters the microscope incubator. It is humidified using a proprietary humidification system that utilizes special tubing that transports water vapor into the gas stream (without changing the CO2 concentration). This humidification system is unique to In Vivo Scientific, and is superior to any other humidification systems available; it significantly increases the water saturation of the gas compared to "bubble" systems (>99% saturation vs. ~80% saturation). This leads to almost no evaporation from the sample over prolonged periods, as well as reduces the chance of contamination (since the gas is not directly in contact with water at all). The humidified gas then passes through a device that sits on top of the microscope stage and equilibrates the gas temperature to the exact temperature of the stage (note that a gas stream is cool upon entry into a chamber; no other manufactures equilibrate gas stream temperatures prior to entry into the sample). Finally, the gas enters a mini-chamber that fits into our stage adapter. This mini-chamber can be custom designed to fit any sized dish/plate as well as stage. It features an optical-grade glass lid suitable for DIC applications, and is designed to fit under all long working distance condensers. Glass lids can be customized for users who require open access for micromanipulators etc. The constant, slow-flow of gas ensures that the sample is surrounded by humidified, defined CO2 environment.
Humidity

IVS-3001:
This system allows for more customization and a higher precision in CO2 control for more demanding applications, or for labs that need more flexibility than pre-mixed gas cylinders can provide (e.g. multiple cell lines with different CO2 requirements). This system requires a 100% CO2 cylinder as well as an ambient air gas supply (via pressurized cylinder or a house ambient air house line). The stream of ambient air passes into the controller at a defined rate (40 mL/min). This air then enters a mixing chamber, into which a second stream of 100% CO2 enters via a variable flow, feedback controlled valve. The user can define the precise percentage of CO2 desired through a digital controller, and change the percentage of CO2 at any time. This gas mixture then leaves the controller and enters the heated microscope chamber. As in the first system, it is humidified using our special tubing humidification system and the temperature is equilibrated to the stage temperature. It then enters the same mini chamber that surrounds the sample. In many ways, this controller is similar to other CO2 systems, though we have chosen to use a stream of ambient gas instead of a fan for a more precise control of CO2 concentration based on empirical testing. Again, no other system is able to achieve such high saturations of water vapor or precisely equilibrate gas temperatures prior to entry into the sample chamber as the In Vivo Scientific chambers.

IVS-3003:
The most advanced CO2 system available for live cell microscopy. Unlike any other CO2 system currently available, the IVS-3003 directly measures CO2 concentration at the sample, and adjusts the stream of incoming CO2 via feedback control to maintain the user's desired CO2 concentration set point. While such a system is inherently more complex than systems that rely on measuring CO2 concentrations prior to reaching the sample (such as the competitor's CO2 system or the IVS-3002), it is the only way to ensure that the CO2 concentration at the sample is actually what the user thinks the is actually what the user thinks the concentration is. This is especially important when manipulations of the sample are being performed (e.g. adding drugs) that might temporarily disrupt the CO2 environment, or when a sample chamber with openings is required for micromanipulation (e.g. cell injections, IVF). Like the other CO2 systems from In Vivo Scientific, the IVS-3003 utilizes specialized humidification tubing and gas temperature equilibration.

Coverslip Chamber:
For high N.A. live-cell imaging, cells must be plated on coverslips or special imaging dishes. Because of the relative cost of coverslips (pennies each) compared with imaging dishes (dollars each), we have designed a tool-less coverslip chamber that is easy to use and quick to assemble (see below). Unlike most other coverslip chambers, ours is round, so the orientation of cells within the field of view can be changed by simply turning the chamber. Made of high-grade stainless steel, the coverslip chamber is fully autoclavable. We suggest the purchase of two coverslip chambers, which allows for pre-equilibration of one sample within the microscope incubator while another is being imaged, expediating live cell imaging experiments.

Perfusion:
If media exchange is necessary (e.g. during tissue slice imaging), we offer a mini-peristaltic pump with perfusion arms/needles that mount directly onto the coverslip holder stage adapter. This perfusion pump has variable speed settings that allow the user to set the desired rate of flow.

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