brochure

Fluorescence Light Sources
Fluorescence Illumination
Mercury, Xenon and LED
A Ray of Light
2
MOOD IMAGE 1
CONTENTS
THE LIGHT OF LIFE
Pushing research forward
The advent of fluorescence microscopy has enabled all researchers to see greater levels of detail more
clearly than ever before. For all processes, no matter how complex, the growing range of dyes and
markers has allowed research into life processes to surge ahead, moving microscopy from being static
to dynamic. New dyes have extended the range of normal fluorescence microscopy, enabling more indepth observation over extended time periods of days or even months. They have also led to a number
of unmatched advancements in Olympus microscope optics, such as the new UIS2 objectives and fluorescence filter sets. The addition of a range of digital cameras, various imaging software platforms and
other modules enables Olympus to provide the correct system solutions, whatever the requirements.
Arc burner light sources
4–7
Arc burners have been the mainstay of fluorescence illumination in microscopy
for a long time and a range of improvements have been implemented to ensure
that they are still excellent options for the majority of procedures. Arc burners
use mercury (Hg) or xenon (Xe) to provide bright light across a broad spectrum
of wavelengths. Recent developments have greatly improved functionality,
including alignment-free bulbs and increased lifetime.
LED light sources
8–11
LEDs have proved themselves as excellent alternatives to standard bulbs in
many lighting situations and have now also entered the microscope fluorescence light source arena with outstanding properties, such as their compact
size, stable alignment-free light and extended life. Olympus has two LED systems, one providing routine transmitted light facilities and the other designed for
more advanced reflected light processes. Both are easy to control and provide
flexible fluorescence.
Real-time illumination systems
12–13
Real-time microscopy needs fast illumination control to capture short-lived
events. Therefore Olympus has developed a range of specialised illumination
systems which provide bright and stable fluorescence excitation light from arc
burners, which are easy to control ‘in tune’ with your experiments.
Your Vision: Our Future
Olympus is dedicated to providing the best combination of microscope components to support your
work at all levels. We have therefore developed a comprehensive range of fluorescence illumination
sources to make the most of any application, whatever the microscope.
4
CHAPTER I
ARC BURNERS
Let there be light
Arc burners are accepted as the standard source of fluorescence for microscopy. Olympus has
enhanced the design of its burners to provide a number of improvements, including a reduced
chance of incorrect use, which provides higher data fidelity, better stability and functionality.
These ensure that researchers can concentrate on their work rather than on the light source.
ARC BURNER LIGHT SOURCES
BURNERS AND BEYOND
Every component of a fluorescence microscope’s light path is important to
the overall image produced. Probably the single most important optical component, though, is the objective lens, since both the fluorescence illumination
and emission pass through it. The new UIS2 objective range from Olympus
uses new glasses to not only provide the best resolution and clarity, but also
to significantly reduce the autofluorescence, consequently greatly improving
the S/N ratio. Furthermore, carefully designed anti-reflection coatings provide
broad transmission width and outstanding chromatic aberration correction for
reduced focus shift. The UIS2 optical system also features excellent fluorescence filters with similar benefits, which enable steep spectral flanges. These
improvements provide the entire light path with a 10–20% increase in efficiency
and an accuracy of ±2 nm.
A
UIS2 objectives
Lead-free optics
B
Fluorescence filter cubes
In various colours
With each optical system providing the best characteristics for the research being
conducted, Olympus has a range of different fluorescence arc burner solutions to
perfectly complement each microscopy system; from lamphouses to fully integrated
units. Olympus has continued developing these solutions to offer the best illumination for all protocols. For example, improvements to the collector lens maximise light
intensity, and Olympus lamphouses ensure that lamps are simple to fit and easily
aligned.
The arc burner-based illumination systems available from Olympus also provide other
enhanced features to ensure that the fluorescence illumination is taken to a higher
level. The X-Cite 120 range and MT_10D modules feature more precise control via
the
family of imaging and analysis software, advanced electronic power control
for longer burner life, as well as integrated intensity (attenuation) and colour filters.
Furthermore, the X-Cite 120 range uses a specialised metal halide technology, to further increase lamp life whilst keeping the same wavelength spectrum.
Building on the MT10_D system, the MT10 and MT20 systems provide an excellent balance of speed, flexibility and brightness, and can use different burners for
expanded research possibilities. For the ultimate in speed, though, the Polychrome V
grid monochromator provides the fastest wavelength changes across its spectrum in
less then 400 ms.
D
Fluorescence light path
On a BX51 microscope
C
Fluorescence lamphouses
With Mercury or Xenon burner
6
A
X-Cite 120PC
Metal halide fluorescence
illumination system
X-Cite 120
A The X-Cite 120 XL range will provide the same fluorescence spectrum and
similar intensities as a standard 100 W mercury burner, but will ensure an
additional level of consistency and safety, making them excellent options for a
broad range of requirements. Furthermore, the units are thermally uncoupled
from the microscope, with a liquid light guide delivering the light through collimators to the Olympus BX2, IX2, SZX2 or MVX microscopes.
Three times better
B
X-Cite interface
In
software
B The three versions of the X-Cite 120 XL range provide each user with the correct
solution for their fluorescence illumination requirements. Each of the three brings the
same level of consistency and safety, but with different levels of controllability. The
entry level model offers a consistent intensity level and is controlled directly from the
unit. The X-Cite 120 XL Iris model adds to this with five intensity levels set by an iris
system (0%, 12%, 25%, 50% and 100%), enabling finer control for sensitive and
bright samples. The X-Cite 120 XL PC combines all these features with PC-based
control via an RS232 interface.
Unique longevity
C The X-Cite 120 XL range uses alignment-free metal halide burners, making them
much easier to handle and move between different microscopes. By using metal
halide technology, much of the tungsten eroded from the electrodes during ‘burning’, which is normally deposited on the inner bulb surface, is recycled back to the
electrodes. This slows down the widening of the arc gap, which in turn decreases
the rate of intensity reduction. This, coupled with the electronic control gear (ECG),
which ensures that as the gap between the electrodes grows the correct voltage is
used to generate a consistent arc, greatly extends the life of the burner and guarantees at least 2,000 hours of use.
C
Pre-aligned burner
For X-Cite 120
Degradation
Typical lamp degradation for X-Cite 120 XL
90%
80%
70%
60%
50%
40%
30%
20%
10%
0%
D
0
200
400
600
800
1,000
1,200
1,400
1,600
1,800
2,000
Elapsed time (hours)
Collimator
For X-Cite 120
Safety first
The unique IntelliLamp™ system monitors and maintains the optimum lamp temperature to ensure consistently safe operation and even prevents the lamp from being
turned on if it has not had time to cool down. Furthermore, it automatically tracks
the lamp’s usage time – importantly, this information stays with the lamp, even if
the lamp is moved to a different unit. The ECG system also ensures that once the
voltage used to maintain the arc reaches a certain level it is shut down safely. This
greatly decreases the risk of dangerous lamp explosions.
ARC BURNER LIGHT SOURCES
MT10_D
A
MT10_D
Advanced fluorescence illumination
A The Olympus MT10_D features all the components necessary for advanced
multicolour fluorescence excitation illumination. It includes the burner compartment, an 8-position filter wheel, 7-level light attenuator and high-speed shutter.
This versatile all-in-one system can be controlled conveniently via the manual
controls or smoothly integrated into the different Olympus
imaging software programmes.
Convenient and easy
A The intuitive manual control can be easily configured and incorporates a digital
status display which makes all of the functions very convenient and straightforward.
Further unique tool-free features simplify other processes such as excitation filter
changes, lamp exchange and intensity optimisation via an integrated photodiode, all
of which also reduce downtime during maintenance.
Fast and flexible
Modern cell labelling techniques enable researchers to use multiple fluorescence
dyes and proteins to provide simultaneous observation of different cellular structures
and physiological parameters. This is made possible by the MT10_D due to the fast
internal 8-position filter wheel, which switches between different filters in just 85 ms.
The 7-level attenuator provides quick adjustments in light intensity and the highspeed shutter can be switched off and on in less than 5 ms, providing fine control
over the light entering the microscope optics.
B
150 W Burner
Xenon or mercury-xenon
Bright and stable
B C The peerless Olympus MT10_D can use either a 150 W Xenon or a 150 W mercury-xenon burner, depending on the wavelength requirements of the research work.
The advanced electronics ensure that both burner sources provide bright stabilised
light over long periods of time, ensuring the utmost data reliability and consistency.
Light is transferred to the microscope via a quartz fibre coupling, which guarantees
the highest levels of light efficiency.
C Highly stable light output
Comparison with standard Hg
lamphouses: light output
normalised to the average
intensity (100%).
Relative
intensity (%)
110
Control and upgrade
MT10/20
MT1
MT
MT10
M
T1
T
T10
10
1
0/2
//20
20
2
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HgH
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bu
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rne
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err
Standard
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Stan
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an
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ard
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bu
bur
urn
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105
The MT10_D can be controlled from a PC using the
family of advanced software packages via RS232 or USB hardware communication interfaces. The MT10_D
can be upgraded to the MT10 or MT20 real-time illumination systems to provide
even better parameters for live-cell imaging. For more about the MT10 or MT20 illumination systems, see chapter 3.
100
95
90
0
5
10
15
20
Time (min)
8
CHAPTER II
LED LIGHT SOURCES
The future is bright and colourful
LEDs have been around for many years, finding new applications as the technology advances.
As a result, LEDs can now be used as alternative sources of fluorescence illumination for
microscopy. Importantly, the most recent advances have made them an even more attractive
solution: from extremely long lifetimes with very little drop in intensity, to high-speed electronic
control and completely stable illumination. LEDs are also very safe and reduce the requirement for intensity filters. All in all, LEDs offer a new level of functionality, bringing fluorescence
microscopy within reach of an even wider range of applications.
LED LIGHT SOURCES
A NEW KIND OF LIGHT
With Olympus microscopes already pushing the boundaries of fluorescence
microscopy with excellent optical systems, the introduction of light emitting
diode (LED) -based fluorescence light sources further expands the possibilities. LEDs use solid-state electronics to emit light within very tight wavelength
bands. This enables more precise illumination of fluorescent dyes with reduced
bleed-through and also simplifies control since one LED can be used for each
specific illumination wavelength range. LEDs are therefore well placed to maximise the impact of new fluorescent dyes. Furthermore, LEDs offer a very long
lifetime (guaranteed minimum of 10,000 hours) with excellent lumen maintenance. This means that during an experiment (however long), the light intensity
will hardly change, which means that LEDs provide highly reproducible results
and the ability to be fully quantitative. This is also useful for screening processes where illumination is key to consistency.
An important aspect, especially in fluorescence live-cell imaging applications, is the
protection of the specimen. The tight wavelength bands emitted by LEDs make it
easier to avoid the dangers of overexposure to light: phototoxicity of the specimen
and photobleaching of fluorescent dyes. Moreover, LEDs do not emit heat in the form
of light, thus the risk of specimen damage by IR radiation is reduced.
Arc burner units use shutters to control when samples are illuminated, whereas LEDs
can be switched on and off very quickly, which provides more controllable illumination windows (into the microsecond range) and therefore also reduces unwanted illumination. LED-based light intensity is controlled uniformly by varying the voltage and
therefore there is no need for neutral density filters as with arc burners. With fewer
mechanical components such as shutters and filter changers, and the flicker-free
nature of LED light, image stability and ease of use are both improved.
LEDs are mechanically very stable and also pose no threat of explosion or exposure
to poisonous gases, which is a great benefit, especially for inexperienced users.
Environmentally, LED-based fluorescence illumination systems offer lower power
consumption due to the increased efficiency of LEDs, reduced reliance on mechanical components and there are no disposal issues. This also means that they have
lower running costs.
A
LED lamp
B
Typical construction
Of a high-performance LED
Lens
Gold wire
LED chip
Body
Cathode lead
Anode lead
Heatsink slug
Silicon sub-mount chip
10
A
FluoLED
CX41 with multi-fluorescence
version
FluoLED
The FluoLED system brings both flexibility and value to LED-based fluorescence, making it perfect for routine and educational requirements, but with the
added benefit of ‘in-field’ operation. The system has been specifically designed
for use with transparent specimens, such as bacteria and thin sections, which
require transmitted fluorescence illumination. The system provides very good
signal-to-noise ratios (S/N) with high intensities and is very easy to retrofit onto
the Olympus CX upright microscope range.
All-rounder
B
Single fluorescence unit
A The entry-level FluoLED fluorescence excitation illumination system introduces
additional flexibility to fluorescence microscopy. It is designed to fit Olympus CX
microscopes making it ideal for educational purposes. Further extending this application area is the ability to power the LEDs by battery or even solar power, ensuring
that fluorescence microscopy can be conducted ‘in field’ where samples are fresh.
Illuminator with LED cassette
Straightforward
B C D The FluoLED system is available in three versions, all of which attach directly to the microscope to provide transmitted fluorescence, without affecting the use of
the normal transmitted brightfield illumination: the FluoLED EasyBlue (480 nm) provides a single wavelength of fixed intensity for the most cost-efficient solution. The
FluoLED Single provides users with the capability to use one of seven interchangeable LED cassettes from 365 nm (UV) to 630 nm (red) and control the intensity of the
light. The FluoLED MultiFluo enables up to three of the interchangeable LED cassettes to be installed and controlled on the microscope at once via a three-channel
electronic driver.
C
Multi-fluorescence unit
Illuminator with three LED cassettes
Simple to use
A With no lamp alignment requirements or the need for excitation filters, the simplicity of the FluoLED systems makes them easy and intuitive to use. An integrated
UV-blocking filter and a filter slider, which holds the emission filters for signal detection, ensure that the systems are safe for the user’s eyes.
A long time going
D
FluoLED
LED cassette
D With a minimum lifetime of 30,000 hours and a 24-month warranty on each LED
cassette, the FluoLED systems are not only cost-effective to purchase, but also very
efficient to run. The low power usage also ensures that fluorescence microscopy is
not a drain on electrical supplies when they are scarce, and that they can be used in
the field from non-mains sources (excluding the MultiFluo).
LED LIGHT SOURCES
precisExcite
A
precisExcite
Easy adaptation to all Olympus
fluorescence microscopes
The precisExcite is a peerless LED fluorescence light source, featuring the latest advances in the LED technology and delivering the most comprehensive
range of wavelengths with the highest available intensities.
Technically advanced
A It is sometimes said that light-emitting diodes do not produce heat. But they do!
Even the best LEDs available today transform most of the applied electric current
into heat and not into light. And LEDs don’t emit heat in the form of IR light. Efficient
cooling is therefore a must to avoid detrimental effects on LED performance such
as reduced intensity, spectral shifts and reduced lifetime. The precisExcite is the
only LED illumination system on the market to use specially developed, proprietary
LED array design and a Peltier cooling system which enable it to attain intensities
not possible on other systems. This also helps to increase the life and stability of
the LEDs to provide in excess of 10,000 hours of use with no more than a 7.5%
decrease in light intensity.
Rapid modularity
B
B The LEDs are provided as LED array modules (LAMs) which are easily interchangeable and enable up to three different wavelengths to be used. The precisExcite is supplied with LAMs for 400, 465 and 525 nm wavelengths – perfect for DAPI,
FITC and Rhodamine dyes, for example. The LAMs are automatically recognised
by the precisExcite so changing between different modules is both quick and easy.
Each LAM has a three-year guarantee and it is therefore possible to reduce running
costs to an absolute minimum.
Installation and control
precisExcite LAM: The brightest fluorescence LEDs currently available
The output from the precisExcite is designed for reflected light microscopy and is
delivered to the microscope via a liquid light guide, which only needs to be aligned
once at installation. The combination of the advanced LED system, liquid light guide
and collimator ensures that the precisExcite provides even illumination across the
entire field of view. The use of LEDs means that there is little need for intensity filters
since the unique electronic control system enables intensity settings from 0–100% in
1% increments.
C The precisExcite can be controlled in two ways. The remote pod provides simple
button functionality along with a clear screen. For a more integrated system, the
precisExcite can be fully operated via the Olympus
family of software programs. This also extends the functionality, enabling the use of TTL pulses to provide
extremely fast on/off switching, which also removes the need for a shutter in the light
path. The software also enables the user to switch between the wavelengths in under
10 ms, as well as store and reuse the parameters used for illumination. This, along
with the excellent stability of the light source, even over long periods of time, ensures
that any data generated is consistent and repeatable.
C
precisExcite hand switch
Intuitive illumination control
12
CHAPTER III
REAL TIME
Don’t make your research wait
At the molecular level, a second is a long time and therefore if you are looking for subtle
changes over time in a cellular system, then you will need to record in real time – see the
changes as they happen, record more data and get the whole picture. Achieving this
requires some dedicated equipment such as high-speed illumination sources with computercontrollable components. The Olympus range of real-time illumination sources takes this a
step further with exquisite precision, excellent reliability and an easy-to-use interface.
REAL TIME
THE NEXT LEVEL IN CELL IMAGING
Advanced live-cell imaging applications, such as multicolour time-lapse imaging, Z-sectioning, multidimensional imaging, ion imaging, FRET, TIRF, etc., need
illumination systems with bright light for efficient illumination, fast switching to
different wavelengths, high-speed shutters for maximum specimen protection
and high precision for analysis. The Olympus MT10 and MT20 light sources provide highly stabilised and easily controlled light from either a 150 W
xenon or 150 W mercury-xenon burner. To maximise the potential of these
unique units, they are fully integrated with the Olympus imaging systems
and
respectively.
A
Screenshot of
software
MT10
B The MT10 (and
) is designed for fast events and incorporates an 8-position
filter wheel which can be moved between neighbouring filters in just 85 ms. Intensity
can be controlled even more quickly via the 7-position attenuator (4–100%) and the
high-speed shutter can be opened or closed in under 5 ms. The
system also
incorporates a system coordinator (PC control board) which provides 10 ms temporal
resolution (camera 1 ms) with a timing precision of 15 ms. The combination of the
MT10, system coordinator and
software makes even the most complex data
acquisitions straightforward and repeatable.
B
MT10
Fluorescence illumination system
MT20
C The MT20 (and
) is designed especially for high-speed live-cell imaging and
uses a faster filter wheel than the MT10 which moves between each of the 8 neighbouring filters in just 58 ms. The expanded attenuator set includes 14 positions
offering intensities between 1 –100%. The shutter is also much quicker, with on/off
times of less than 1 ms. The more powerful real-time controller PC board ensures,
via parallel command execution, temporal resolutions of 1 ms and precision in the
microsecond range.
C
MT20
Fluorescence illumination system
for high-speed imaging
Experiment Manager
With both the
and
software programs, the powerful Experiment Manager
provides precise planning and execution tools via an intuitive drag-and-drop system
that enables easy assembly of command icons such as image acquisition,
Z-stack or time loop.
Spectral microscopy
D For the ultimate in speed, the
system can be used with the Polychrome V
monochromator-based system, which can provide wavelength changes of up to 400
nm/ms. The Polychrome V uses a 150 W xenon burner with 3,000 h lifetime and provides UV to near-infrared wavelengths. An optional 10-level attenuator enables fast
setting of different excitation light levels.
D
Polychrome V
Ultimate wavelength switching
speed
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Light sources specifications
Arc burner light sources
Mercury lamphouses
Xenon lamphouses
X-Cite 120
X-Cite 120 Iris
X-Cite 120 PC
Type
Mercury
Xenon
Metal halide
Metal halide
Metal halide
Wattage
50/100
75
120
120
120
Lifetime (h)
100/300
1,000
2,000 (guaranteed)
2,000 (guaranteed)
2,000 (guaranteed)
Operation
Manual
Manual
Manual
Manual
Manual/PC
Intensity control
Additional ND filter
Additional ND filter
Additional ND filter
Shutter: Manual
Online
Automatic
Iris with 4 positions plus
Iris with 4 positions plus
shutter function
shutter function
Via slider in illuminator
Via slider in illuminator
Via slider in illuminator
Via iris
Via front panel/foot switch
–
–
–
–
Online via GUI
–
–
–
–
Via SW
NA
NA
NA
NA
≥ 200 ms
Filter positions
NA
NA
NA
NA
NA
Switching speed
NA
NA
NA
NA
NA
Control interface
NA
NA
NA
NA
RS232
Real-time system
NA
NA
NA
NA
No
BX, IX, MVX, SZX (100 W)
BX, IX, MVX, SZX
BX, IX, MVX, SZX
BX, IX, MVX, SZX
BX, IX, MVX, SZX
Shutter speed
Microscope
CX and CKX (50 W)
MT10_D
MT10
MT20
Polychrome V
Type
Mercury or mercury-xenon
Mercury or mercury-xenon
Mercury or mercury-xenon
Xenon
Wattage
150
150
150
150
Lifetime (h)
1,000
1,000
1,000
3,000
Operation
Manual/PC
PC
PC
PC
Intensity control
Attenuator with 7 levels
Attenuator with 7 levels
Attenuator with 14 levels
Attenuator with 10 levels (optional)
Shutter: Manual
Via hand switch
–
–
–
Online
Online via GUI
Online via GUI
Online via GUI
Online via GUI
Automatic
Via SW
Via SW
Via SW
Via SW
Shutter speed
< 5 ms
< 5 ms
1 ms
< 1 ms
Filter positions
8
8
8
Continuous wavelength selection (320 – 680 nm in 1 nm increments)
Switching speed
≥ 85 ms
≥ 85 ms
≥ 58 ms
Up to 400 nm/ms
Control interface
R232/USB
Independent plugin CPU board
Independent plugin CPU board
Voltage (–10 V to + 10 V), RS232, trigger in and out
Real-time system
–
–
Yes
Yes
Microscope
BX, IX, MVX, SZX
BX, IX, MVX, SZX
BX, IX, MVX, SZX
BX, IX, MVX, SZX
LED light sources
FluoLED Single Fluorescence
FluoLED MultiFluo
precisExcite
Illumination method
Transmitted fluorescence
Transmitted fluorescence
Transmitted fluorescence
Reflected fluorescence
Lifetime (h)
10,000 (minimum)
10,000 (minimum)
10,000 (minimum)
10,000 (minimum)
Wavelength
480 nm fixed
One
Up to three
Up to three
Available LED modules
Blue (fixed)
UV/royal blue/blue/cyan/
UV/royal blue/blue/cyan/
10 different LAMS
green/yellow/red
green/yellow/red
From 400 to 635 nm
Intensity control
Fixed
One-channel electronic driver
Three-channel electronic driver
0 –100% in 1% increments via pod or SW
Cooling
NA
NA
NA
Peltier
Control interface
NA
NA
NA
RS232/Ethernet/USB
Battery pack for in-field use
Optional
Optional
NA
NA
Microscope
CX21/31/41
CX21/31/41
CX31/41
BX, IX, MVX, SZX
The manufacturer reserves the right to make technical changes without prior notice.
Postfach 10 49 08, 20034 Hamburg, Germany
Wendenstrasse 14 –18, 20097 Hamburg, Germany
Phone: + 49 40 23 77 30, Fax: + 49 40 23 77 36 47
E-mail: [email protected]
www.olympus-europa.com
Art. code: E0430944 • Printed in Germany 04/2008
FluoLED EasyBlue