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FCS2 Closed Chamber System

FCS2 & FCS3 Controller

FCS2 Stage Adapter
(Olympus Adapter Shown, Other Stage Adapters Available)

FCS2 Chamber
The system is comprised of:

Electronic Controller
Stage Adapter
Chamber (environmental optical cavity)

What is it?

The Focht Chamber System 2 (FCS2 ) is a closed system, live-cell micro-observation chamber, that offers several advantages over other chambers. In addition to its unique perfusion and thermal control systems 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 an air curtain.

Why do you need an FCS2?

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 can provide low shear near laminar flow as well as high shear narrow directed flow and anything in between. The most important feature of the FCS2, other than unequalled temperature uniformity, is that there is a precise directed flow of media over the cells. Open dish type chambers do not provide the flow capabilities of the FCS2. The FCS2 provides an optical imaging cavity where the user can precisely define the optimum flow characteristics necessary for the experiment.

FCS2 users enjoy the following characteristics:

  • Easy no tools needed assembly
  • Compatibility with all modes of microscopy
  • Complete control of the volume within the optical cavity
  • Complete control of the separation between optical surfaces
  • Complete control of the shape or profile of the flow channel
  • Complete control of the temperature uniformly across the entire field
  • The FCS2 is the most recognized and published commercial cell chamber available


FCS2 Demonstration Video
Click to view, QuickTime 7 required, 30mb

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 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.

Temperature Control:

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.

Thermographic Images:

Visible Light Image of a Bioptechs FCS2

Thermal Profile of a Bioptechs FCS2

The thermograph above demonstrates the uniform temperature distribution of an FCS2. 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.

Microaqueduct Perfusion:

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)

FCS2 Optical Cavity Half Section

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 no flow through high rate 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
  • Closed system so that bicarbonate CO2 or organic buffers can be employed
  • Compatible with 1/16" tubing for perfusion (C-Flex, Tygon, etc.)
  • Easily assembled with ordinary skill (no tools required)
  • Stand-alone temperature controller with an alarm circuit to safeguard your cells
  • Near laminar flow
Standard FCS2 Chamber Specifications:
Physical Size 75mm OD 13mm high
Coverslip No. 1.5 thick x 40mm Diameter
Imaging Aperture 22mm
Maximum Volume 706mm^3
Minimum Volume 4mm^3
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


FCS2 Exploded View

  1. Electrical Enclosure
    Can also be detached to sterilize the perfusion tubes
    Temperature sensor
    Heater contacts
  2. Upper Half (Top)
    Contains the perfusion tubes
  3. Perfusion Tubes (14 gauge) (CO2 Perfusion Drawing)
  4. Upper Gasket
  5. Microaqueduct Slide
    An optical surface which integrates perfusion and temperature control
    High-volume laminar flow
    Koehler Illumination
    Electronically conductive coating for temperature control
  6. Singular lower gasket
    This gasket can have any internal geometry you desire
    Standard thicknesses from .1mm to 1mm
    Allows you to define the volume and flow characteristics of the chamber
  7. 40mm coverslip
    Surface where your cells are grown
  8. Self locking base
    Designed to assure parallel uniform closure, eliminate leaks, & broken coverslips
    Temperature controlled
    Dovetail mounted to scope for stability
    No tools for assembly

 

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. See the additional gaskets shapes chart below.

Additional gasket shapes

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


CO2 Enriched Media Schematic for Bioptechs FCS2 & FCS3 systems. Easy to setup, reliable and consumes very little CO2

FCS2 Chamber Flow Cell
One flow cell so universal that it can meet the needs of nearly all applications.

The flow cell is easily modified with a variety of gasket geometries (see additional gaskets above for a few samples), and a gasket thicknesses ranging from 0.1mm to 1mm. If you have your scope in an environmental box or don't need temperature control the FCS2 universal flow cell in the following package provides a substantial savings:

The FCS2 inverted microscope closed system chamber without intrinsic heating capability contains:

1 FCS2 chamber*
5 Microaqueduct slides non-coated
30 piece gasket set
50 coverslips.

* requires stage adapter, see main price-list for model to fit scope

 Open Mode Top for the FCS2

The open mode option allows for the FCS2 to be assembled without the microaqueduct slide thus exposing the cell on the cover slip for microinjection. The coverslip can then be removed and reassembled with the microaqueduct slide for long term, time-lapse. Click here for User Instructions


FCS2 Open Mode Top with cable

FCS2 Open Mode Top installed on FCS2 base ready for microinjection
Low Dead Volume Top for the FCS2

The low dead volume top is specifically designed to reduce the volume of media in the ports of the FCS2 from the point where the supply tubing attaches to the FCS2 through to the optical cavity where the cells reside for imaging. It is commonly used when working with expensive compounds and primers. Note: When using a low dead volume top, the flow of media is greatly reduced by both the aperture and length of the tubes.  The following charts and examples illustrate the differences. The standard FCS2 has a minimal cross sectional area of 0.608 sq mm, the inside diameter of the vertical tube provides ample flow for most experiments. On the order of fractional to milliliters/sec. However, the lowest dead volume top has a minimal cross sectional area of 0.196sq mm. If you do not have high volume flow requirements for your experiment this is not a problem but this must be taken into consideration when ordering.


Low Dead Volume Top


Low Dead Volume Top (bottom)

Half Section Drawing
Standard FCS2 Top:
Tube Tube Length (mm) Tube Inside Diameter (mm) Dead Volume (l)
A 29 1.57 56.14
B 10 0.88 6.08
Total Dead Volume 62.22
 
Low Dead Volume Top 1mm Tubes:
Tube Tube Length (mm) Tube Inside Diameter (mm) Dead Volume (l)
A 29 1 22.78
B 10 0.88 6.08
Total Dead Volume 28.86
Low Dead Volume Top 0.5mm Tubes:

Tube

Tube Length (mm) Tube Inside Diameter (mm) Dead Volume (l)
A 29 0.5 5.69
B 10 0.88 6.08

Total Dead Volume

11.77

Cooled FCS2

CFCS2 Cutaway Rendering with FCS2 Cooling Adapter

CFCS2 with FCS2 Cooling Adapter

How it works:

Cells are plated on a 40mm coverslip and placed into a FCS2 chamber. This provides a perfuseable laminar flow optical chamber with user modifiable flow characteristics. The upper glass element (Microaqueduct slide) is then used to remove heat from the specimen cavity to be absorbed in the cooled fluid being circulated in the cavity formed by the addition of an FCS2 cooling Adapter which is an o-ring sealed window adapter. Gravity or an electric pump is then used to create a flow of chilled fluid through the heat exchange cavity in the chamber (the space in-between the Microaqueduct slide and the glass window of the FCS2 Cooling Adapter). The cells remain safely enclosed in a separate optical enclosure. If more thermal transfer is necessary, tubing can be installed into the FCS2 base (CFCS2 Link).

Note: If you are using high N.A. objectives at below ambient temperatures it will be necessary to cool and thermally isolate the objective from the scope. The optional thermal isolator will prevent condensation from forming on the lower element of the objective. A gas port is provided to purge the adapter with dry nitrogen. An anti-reflection coated window optimized for 340-700nm seals the bottom of the adapter. Cooling Collar and Isolator link.

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 (.pdf)
Micro-Environmental System Profile Questions (.doc)

Accessories:
Objective Heater System Required for immersion objectives
CFCS2 System When below ambient temperatures are required of your flow cell
Objective Cooling Collar To aid your flow cell in below ambient temperatures
   

We wish to acknowledge The Center for Light microscope Imaging and Biotechnology , a National Science Foundation Science and Technology Center, at Carnegie Mellon University, for their initial contribution in the development of the FCS2. The FCS2 is now manufactured in part under license from C.M.U.

E-mail: info@bioptechs.com
Phone: (724) 282-7145, Toll-Free 877 LIVE-CELL (548-3235), Fax: (724) 282-0745