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FRET Split imaging system
• Simultaneous two-color split imaging with one CCD camera.
• Unique design splits the primary image for the highest efficiency
and light transmission necessary for weak fluorescence signals
such as CFP/YFP FRET experiments.
•Compact and space-saving design takes advantage of the 70 mm
of free space between the microscope frame and the primary
image plane found on all Olympus Research Upright and Inverted
Microscopes.
• Simple cassette mechanism makes it easy to switch between
split and full frame imaging.
• Unit is up to 10% brighter than similar relay lens based, image
splitting systems.
• When used with the rectangular field stop U-RFSS, excitation
energy is limited to the camera's field of view, minimizing specimen
photo-bleaching.
Bright, simultaneous two-wavelength imaging using the primary image.
Cube cassette for
full image
Rectangular field
stop/U-RFSS
Split primary image
camera port/U-SIP
Cube cassette for
split images
Prism
Built-in separation
dichromatic mirror
(DM505)
Filter sliders
(Emission, ND sliders)
Filter set such as
XF88-2(OMEGA) or
31044v2(CHROMA)
Specimen
Objective
Excitation filter
Tube lens
75W xenon apo
lamp housing
U-LH75XEAPO
Power supply unit
Research inverted
system microscope
IX81/IX71
L shape fluorescence illuminator
IX2-RFAL
Fluorescence mirror unit
Split primary image
camera port
U-SIP
Cube cassette for
split image
Mirrors
Split image
High resolution camera
YFP
CFP
Rectangular field stop
U-RFSS
U-SIP main specifications
Microscope
IX71/81
Image separation
Right and left 2-separation
(can be adjusted independently)
Built-in separation dichromatic mirror
DM505 (special size)
Filter slider
Emission, ND filters' size ø25 mm,
total thickness: 8 mm
Used together with commercially available filter set
(XF88-2 OMEGA) or 31044v2(CHROMA)
Field Number
Split image: 8
Full image: 11
Magnifications
1X (primary image)
Objectives
40X and higher
Camera mounting
C-mount
Recommended camera
Chip size 2/3 inch
YFP
CFP
HeLa cell, in which YC3.1 (cytoplasm) and YC3.1nu (with nuclear localization signal) are coexpressed.
FRET changes are observed through histamine stimulation, and images are acquired at intervals of
50 msec.
FRET
Photoactivation Fluorescence
Microscope system
The photoactivation illuminator allows the exposure by UV light to
specific regions of a cell for photoconversion, the uncaging of
compounds and the photoactivation of specific fluorochromes.
• A specified area of the cell can be exposed to UV light while
observing the targeted cell by fluorescence or transmitted (DIC)
method.
• Compliance with FRAP or FLIP
experiments (by special order).
• Easy system upgrade by
attaching double lamphouse
illuminator IX2-RFAW to IX2
series inverted microscope.
Photoactivation illuminator for inverted microscopy.
The novel Kaede gene is useful in biology because it exhibits photoconversion. Normally, the Kaede
gene shows green fluorescence but after exposure to UV light will exhibit red fluorescence. By using
UV light to only a specific region within a labeled cell and then noting the movement of red beyond
that region, observations of internal cellular dynamics can easily be made. The photo on the left
shows a nerve cell (from a rat hippocampus) pre-labeled with green Kaede gene.
The photo on the right side was taken after the right-most cell body was exposed to a 10 µm
diameter spot of UV light for 60 seconds, thus changing the Kaede gene from green to red. Note the
translocation of the red shifted gene outside of the 10 µm spot thus indicating intracellular transport
mechanisms.
High resolution
camera
Excitation filter
BP330-385
Filter slider
Composed dichromatic
mirror of right and left path
Excitation filter
475AF20(XF1072 OMEGA) or
HQ475/20x(CHROMA)
Excitation filter *2
550DF30(XF1021 OMEGA)
Fluorescence mirror unit
U-MF2+dichromatic mirror
(DM400 on the illustration)
Objective
UPLFLN40XO
Filter wheel 2
such as Lambda10-2
Fluorescence filter *2
575ALP (XF3089 OMEGA) or
HQ575LP(CHROMA)
Fluorescence filter
530DF30(XF3107 OMEGA) or
D530/30x(CHROMA)
Filter slider
Inverted microscope
IX series
Pinhole slider
Pinhole or slit
Magnetic shutter
Sample
Filter wheel 1
such as Lambda10-2
Double lamp
house illuminator
IX2-RFAW
Field
stop
position*1
Double lamphouse illuminator IX2-RFAW
IX2-RFAW specifications
Microscope
IX81/71/51, IX70/50
Pinhole slider
2-step exchange (pinhole or slit/vacant hole)
Pinhole and slit are available on the market
(ø16 mm Melles Griot Inc. products)
Exposed area on
Pinhole diameter
the specimen
objective magnification
Filter slider
3-step exchange (shutter/filter pocket/vacant hole)
BP330-385 excitation filter equipped
Excitation filter slider
5-step exchange (4-step filter pocket/vacant hole)
Filter size
Excitation filter: ø25 mm,
thickness: 6 mm and below
ND filter: ø32 mm,
thickness: 1 mm and below
Composed dichromatic mirror
DM400 (standard)
of right and left light path
Slide IN/OUT type
Power consumption
7.4 A
Dimensions
Width: 710 mm
Depth: 740 mm (from the front of tilting tube to
the end of the illuminator)
*1 Field stop position is the same
position as the focus point of the
sample.
*2 Use 550DF30 excitation filter in the
filter wheel 1 and 575ALP
fluorescence filter in the filter wheel 2
when observing red Kaede protein.
Exchange of the fluorescence mirror
unit is not required.
Photoactivation
Application System
* Not available in some areas
* Not available in some areas
Setting up example for Kaede
