The FCS2® System is a closed system, parallel plate flow cell environmental chamber used on a microscope to enable the acquisition of high resolution, time-lapse images of cells undergoing either natural growth or fluidic exposure to variant factors. The FCS2® System is designed to be used on any inverted microscope as the FCS3® System is specifically for upright microscopes. Both systems enable the user to define precise and unlimited flow characteristics within the chamber to suit their protocols. Bioptechs’ patented thermal design provides uniform temperature control across the entire field. This is a feature that is not possible with a peripheral only heated chambers.
What is it?
The Focht Chamber System 2 (FCS2®) is a closed system, live-cell micro-observation chamber that utilized several patented technologies that offer several exclusive advantages over other chambers. In addition to its uniform temperature control and user definable perfusion capability, it is fully compatible with all modes of microscopy. It is also the only chamber to combine high-volume laminar flow perfusion rates with Koehler illumination and precise temperature control without the need for an air curtain. This is accomplished by the development of micro aqueduct perfusion. A method of introducing media into a fluid optical cavity where the separation of optical surfaces and flow geometry is definable by the user. Therefore, the chamber is adaptable to the protocol instead of having to adapt the protocol to the chamber.
The FCS2 is a flow cell or flow chamber that is designed specifically for the demands of today’s live-cell imaging requirements. It has limitless flow characteristics because its flow geometry can easily be customized by the user. It provides the ability to configure the optical cavity to produce control shear forces, flow characteristics, high or low volume exchange rates, near laminar flow, and anything in between. The most important feature of the FCS2, other than unequalled temperature uniformity, is that the is flow of media is constrained to a precise location over the cells. Open dish type chambers do not provide the flow capabilities of the FCS2. The FCS2 provides unmatched, uniform temperature control, in a user configurable, perfusable, optical imaging cavity.FCS2 users enjoy the following characteristics:
How does it work?
Cells are grown on a 40mm glass coverslip. This coverslip is then incorporated into a perfusable fluid optical cavity that is compatible with all modes of microscopy, and its geometry can be easily defined by the user. This optical cavity is secured into a fixture on the stage of the microscope where it can be perfused with media or remain static. Media that comes into one of the ports on the side of the chamber, emerges in a fluid optical path where the media is precisely directed over the cells. The media is collected within the optical cavity and directed out of the chamber on the other side. The flow characteristics of the media while in the optical cavity are easily modified by selecting gaskets of varying geometry that direct the flow. Temperature control is maintained by both peripheral thermal support from the chamber’s metal frame and heat that is emitted from an electrically conductive optically transparent coating on the “window” of the optical cavity. This technique provides uniform temperature control across the entire field, thus eliminating the temperature gradient that occurs with other designs. This technique also enables the FCS2 to recover from perfusion induced temperature fluctuations in just seconds.
The FCS2 was designed to maintain accurate thermal control and allow high-volume laminar flow perfusion. Both of these functions are incorporated into our patented Microaqueduct Slide (see drawing below). The surface of the slide, opposite the specimen side, is coated with an electrically conductive transparent thin film of Indium-Tin Oxide (ITO) and two electrical contacts (busbars). When the FCS2 is completely assembled two electrical contacts, (not shown in drawing), which are contained in the electrical enclosure rest on the busbars. A temperature controller is used to pass a regulated current flow through the ITO Coating. This causes the surface of the slide to heat. The heat is transferred through the perfusable media to the cell surface on the coverslip thereby providing first surface thermal control. The self-locking base of the chamber is also temperature regulated to provide peripheral heat as well.
The thermograph above demonstrates the uniform temperature distribution of an FCS3. Notice that the coverslip temperature is so uniform that its location, in infrared, is indistinguishable from the base of the chamber. This demonstrates the effectiveness of the ITO heated Microaqueduct Slide. It is capable of re-equilibrating cell temperature within seconds of perfusion and eliminates the typical thermal gradient that occurs with peripheral heating.
A fluid pathway is formed by separating the Microaqueduct Slide from the coverslip containing cells with a single silicone gasket. This gasket can be any thickness from 50 micron to 1mm and any lateral geometry you choose or create. This arrangement allows the user to define the flow characteristics. Therefore, you are not limited by the geometry of the optical cavity. Instead you select or create it! Fluid access to this flow channel is made through two 14-gauge needle stock tubes protruding from the sides of the chamber top. These tubes provide fluid connection to two perfusion holes in the Microaqueduct Slide that interface two “T” shaped grooves cut into the inner surface of the Microaqueduct Slide. The “T” groove allows the media to seek the path of least resistance and become nearly laminar before flowing across the cells. This technique eliminates the need for the metal perfusion ring and additional gaskets, which are the limiting factors, required by most conventional chambers. (see drawing below)
Isometric View of Optical Cavity
Microaqueduct Slide is transparent glass but rendered as a solid to improve contrast to illustrate function. Microaqueduct design enables proper Koehler illumination with high-numeric aperture optics for both transmitted and reflected modes of microscopy. Suitable for static or high rate of flow procedures where a rapid exchange of media is required with low cell surface shear Cell temperature can be controlled from ambient to 50 degrees C +/- 0.2 degrees C without the need of an air curtain. Temperature is controlled uniformly across entire field with media equilibrating as it enters the chamber. (see drawing above)
Items 5,6 & 7 are the optical cavity, the gasket (#6) can be changed to any one of the gaskets below or a custom gasket can be made to make any flow geometry, or media volume you want. The coverslip comes in a standard 0.17mm, #1.5 thickness but is also available in 0.5mm, #5 coverslip.
Singular lower gasket
By simply changing this one gasket you can change the volume of the chamber. This gasket can have any internal geometry you desire and can be any thickness from 0.1mm to 1mm. The drawing below shows the standard shapes of the gaskets that we include with every FCS2. We also include solid gaskets for you to custom fit to your application. Once you have found the shape that works best for your experiment, you can contact us to have a die made to those specifications. Examples of standard gasket outlines (below)
Standard Gasket Set includes:
(3) 0.1mm Thick 30mm Round
(2) 0.1mm Thick 14 x 24 Rectangle
(3) 0.25mm Thick 30 mm Round
(2) 0.25mm Thick 14 x 24 Rectangle
(1) 0.25mm Thick Blank
(3) 0.5mm Thick 30mm Round
(2) 0.5mm Thick 14 x 24 Rectangle (2) 0.5mm Thick Blank
(5) 0.75mm Thick 30mm Round w/holes
(2) 0.75mm Thick 14 x 24 Rectangle
(1) 0.75mm Thick Blank
(2) 1.0mm Thick 30mm Round
(1) 1.0mm Thick 14 x 24 Rectangle
(1) 1.0mm Thick Blank
To order a specific geometry please see the price list for part numbers and prices. Gaskets are sold 5 gaskets per pack.
In addition to the standard gasket set that comes with the FCS2 Starter Kit, shown above, there are a variety of gasket geometries, and thicknesses ranging from 0.1mm to 1mm available. Custom thicknesses and materials available upon request. See the additional gaskets shapes chart below.
Custom shapes are available. Simply contact Bioptechs to make arrangements for their production. All dimensions below are in millimeters. Please specify the die number that is located on the gasket drawing below and the thickness of the gasket when ordering.
Perfusion Diagram for FCS2
To assist you and Bioptechs in selecting the most appropriate system to meet your needs, please review and fill out the following questionnaire.
Micro-Environmental System Profile Questions
FCS2 Starter Set includes:
Standard FCS2 Chamber Specifications:
|Physical Size||75mm OD 13mm high|
|Coverslip||No. 1.5 thick x 40mm Diameter|
|Maximum Volume Exchange Rate||1/sec|
|Minimum Fluid Aperture||0.6mm^2|
|Separation between optical surfaces||50 – 1000 microns|
|External port ID||1.6mm|
|Temperature Stability||+/- 0.2 degrees C|
|Standard Temperature Range with use of Electronic Controller||Ambient to 55 degrees C|
Standard FCS2 Top:
|Tube||Tube Length (mm)||Tube Inside Diameter (mm)||Dead Volume (μl)|
|Total Dead Volume||62.22|