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FCS2 Closed Chamber-Inverted
After rigorous preparation your
cells will need a micro-observation environment that is conducive
to their viability, compatible with your experiments protocol
and all techniques of microscopy!
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.
The system is comprised of:
• Chamber (environmental
optical cavity, this page)
• Electronic Controller (this page)
• Stage Adapter
Accessories recommended for immersion objectives:
• Objective Heater
• Objective Temperature Controller
How does it work?
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. 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.
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.
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

Isometric
View of Optical Cavity
Microaqueduct Slide is transparent
glass but rendered as a solid to improve contrast to illustrate
function. This image is rendered upside down, to demonstrate
how the
Optical Cavity works. In an FCS2 chamber the coverslip will be on the bottom.


Assembled FCS2
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) |
| 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 |
|
| 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 |
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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 .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)

The Gasket Set Includes:
(3) 0.1mm 30mm Round
(2) 0.1mm 14 x
24
(3) 0.25mm 30 mm Round
(2) 0.25mm 14 x 24
(1) 0.25mm Blank
(3) 0.5mm 30mm Round
(2) 0.5mm 14 x 24
(2) 0.5mm Blank
(5)
0.75mm 30mm Round w/holes
(2) 0.75mm 14 x 24
(1) 0.75mm Blank
(2) -1.0mm 30mm Round
(1) 1.0mm 14 x 24
(1) 1.0mm Blank
Custom shapes are available. Simply contact Bioptechs to make
arrangements for their production.
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.
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| Open Mode Adapter
Installed on FCS2 Base |
Open Mode Adapter
Installed on FCS2 Base |
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FCS2
Specifications
75mm OD 13mm high
1.5 thick x 40mm
Diameter
:
22mm
706mm^3
<31mm^3
1/sec
0.6mm^2
- 1000 microns
1.6mm
+/-
0.2 degrees C
Important Notes
If you are using immersion
objectives on mammalian specimens, you will need an Objective
Heater!
Click
here to download an assembly and demonstrational video
of the FCS2 (30 MB)
Click here to download a PDF of FCS2 user instructions (724
K)
Click here download a PDF about Traditional Stage Warming with
thermal images. (340 K)
Click here to download a
PDF about Typical FCS2 Perfusion Configuration For Induced Change
Biology (724
K)
To assist you and Bioptechs in selecting the most appropriate system
to meet your needs, please download and fill out a PDF of Micro-Environmental
System Profile Questions (36 K)
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