FluoLED reflected light fluorescence illuminator - · PDF file• Medical and biological ... FluoLED reflected light fluorescence illuminator is available for the following Olympus

For internal use only

FluoLED reflected light fluorescence

illuminator for UPRIGHT transmitted light microscopes with infinite corrected optics

PRODUCT DESCRIPTION

Fraen Corporation Srl For internal use only

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CONTENTS Philosophy and goals……………………………………………………page 3 The new FluoLED reflected light fluorescence illuminator…………. page 5

Where to sell – main applications…………………………………….. page 7 When reflected and when transmitted?............................................page 8 Sales arguments…………………………………………………………page 11 Specifications…………………………………………………………….page 12

Fraen Corporation Srl

Via Stelvio, 12

20019 Settimo Milanese (MI), Italy

Phone: +39 02 335456.1

Internet: www.fraensrl.com

Email: [email protected]


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Philosophy and goals The new FluoLED illuminator for reflected light fluorescence has been developed after collecting

several years of experience in development, manufacturing and marketing of collimating optics for

high power LEDs and optical instruments.

It is intended for universal applications in routine and research fluorescence applications on upright

microscopes.

Similar to the transmitted light fluorescence illuminator, this new unit is available in three different

versions:

“Easy” FluoLED reflected light illuminator

For one LED cassette, with non variable light control, for Royal Blue 450 nm, Blue 480 nm or

Green 535 nm LED cassettes.

“Single” FluoLED reflected light illuminator

For one LED cassette, with variable light control, for UV 365 nm, Royal Blue 450 nm, Blue 480 or

Green 535 nm, Yellow 590 nm and 630 nm LED cassettes.

“Multi” FluoLED reflected light illuminator

For up to three LED cassettes, with variable light control, for UV 365 nm, Royal Blue 450 nm, Blue

480 nm, Green 535 nm, Yellow 590 nm and 630 nm LED cassettes.

Main characteristics are:

1) Reflected light path including a pre-centred field diaphragm for exact sizing of the illuminated

field

2) A 4 positions filter slider for fluorescence filter sets which is completely dust protected avoiding

any contamination from polluting agents

3) Freely interchangeable LED cassettes according to FluoLED version. LED cassettes are

independent and can be retrofitted at any time. They are completely maintenance free and no

alignment is required.

4) Variable light control units allow adjustment of illumination intensity and therefore represent a

helpful tool in preventing sample bleaching.


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In order to increment success in this product segment the following strategies have been

implemented:

Great attention to price/performance ratio.

Analysis and identification of different market segments for FRAEN’s FluoLED illuminators:

when reflected and when transmitted?

Reduction of parts required to fit on microscopes of different brands: “One product fits many

microscopes” – for cost reduction, simplified logistics and easy quoting.

The new FluoLED reflected light illuminator meets theses ambitious goals .


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The new reflected light fluorescence illuminator fr om Fraen

The development of this new product was driven by specific requests coming from target customer

groups.

It does not replace the existing FluoLED transmitted light illuminator. Please refer to section “When

reflected and when transmitted?” for additional information.

“Easy” and “ Single” FluoLED reflected light illuminator:

1) Slider for fluorescence filter sets: typically one position is left free for transmitted white light

illumination (brightfield, phase contrast, …) and three positions are available for

fluorescence filter sets. Each filter set contains an exciter, a beamsplitter and an emission

filter.

2) Filters are completely dust protected by closed housing.

3) Read which filter set/LED is in use. Sticker is in front position to avoid any confusion.

4) Field diaphragm.

5) LED cassette with integrated heat sink.


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“Multi” FluoLED reflected light illuminator:

In addition to the “Easy” and “ Single” FluoLED reflected light illuminators shown before, the “Multi”

FluoLED reflected light illuminator allows to install and drive up to 3 LED cassettes.

1) LED cassette with integrated heat sink. Up to three LED cassettes can be used.

2) Beamsplitters.

Similarly to the transmitted light illuminator, these new reflected light units come with the well known fixed

and variable light controls.

1-colour non variable light control, for “Easy” FluoLED reflected light illuminator

1-colour variable light control, for “Single” FluoLED reflected light illuminator

3-colours variable light control, for “Multi” FluoLED reflected light illuminator


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Where to sell – main applications The new FluoLED illuminator for reflected light fluorescence is indicated for the daily routine applications

in medicine and biology. Typical examples are immunofluorescence, micro bacteriology and basic FISH

investigations.

It can be offered every time a customer wants to retrofit an existing upright transmitted light microscope

with LED fluorescence capabilities.

Please check on section “Specifications” for availability list.

Furthermore also complete microscopes can be offered.

These are the typical requirements fulfilled by the FluoLED illuminator for reflected light fluorescence:

• Upright microscope (new or retrofitted) with all it’s components (objectives, tube,…)

• Routine fluorescence applications in medicine and biology

• Multiple fluorescence (more fluorochromes)

• Good price/performance ratio

• Extreme long life time of LED sources - 30.000 hrs, with integrated heat sink.

The following customer groups are potentially interested in LED fluorescence microscopy:

• Hospital laboratories (immunology, urology, cytology, toxicology, microbiology, …)

• Food industry

• Medical and biological laboratories of any type (also veterinarians)

• Research institutes

• OEM customers

Bacteriology • Tested equipment for widely diffused applications like TB and Malaria • Easy to retrofit on existing microscopes, allows cost reduction in 3rd world countries • Operation on field with optional battery pack Immunology • Detection of autoimmun diseases on standardized test slides like HEp-2, AMCA,

animal tissues. • Detection of single Fitc or Fitc with counterstainig (example Evans blue)

Education

• Robust design, no fiddling with light beams and potential risks from light source explosion (Mercury lamps)

• A selection of microscopes for best price/performance ratio

Basic Research

• Affordable, easy to use system for general laboratory implementation • Ability to perform one, two and three color assays


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When reflected and when transmitted?

FluoLED is available in two different optical layouts:

FluoLED for transmitted light fluorescence

FluoLED for reflected light fluorescence

Each of these two layouts has specific advantages and in order to understand the differences it is

necessary to give some optical explanations.

FluoLED for transmitted light fluorescence

In transmitted light fluorescence, light emitted by the LED cassette is used in transmission mode

rather than reflected mode. Bright field microscopy is not affected since the halogen white light

function remains intact, which means that transmitted light observation is possible without major

changes in the optical configuration.

The kit is designed to attach to a standard bright field microscope and does not change the

characteristics of the microscope in any way.

The adaptor (1) is clamped over the microscope base. The LED cassette (2) is inserted into the

adaptor from the front side. A 45° mirror (3) refle cts light emitted by the LED cassette through the

condenser (4) on the sample.

Fluorescence emitted by the sample is then filtered by an optical filter (not shown) before reaching the

observer.


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In transmitted light fluorescence the condenser creates a specific power density of light on the

sample, which is a fixed value. Objectives with smaller magnification typically exhibit a larger

diameter of the front lens and therefore “capture” more light emitted by the sample. In addition,

economical standard objectives have less lenses inside and this increases internal tranmittance.

FluoLED for transmitted light fluorescence is very well indicated for those applications

where medium to low power objectives are used, typi cally 5x, 10x, 20x up to maximum 40x.

No UV excitation is available.

FluoLED for reflected light fluorescence

In reflected light fluorescence the optical light path is different. This layout is very ofted described

as “epi-fluorescence”, where light is coming from top and the objective has a double function: it

provides illumination on the sample and collects the emitted light.

Obj

ectiv

e

10X

-0.2

5NA

Obj

ectiv

e

40X

–0.6

5NA

CONDENSER

Power Density (W/mm2): same value with allobjectives

CONDENSER

Microscope stage

Obj

ectiv

e

10X

-0.2

5NA

Obj

ectiv

e

40X

–0.6

5NA

CONDENSER

Power Density (W/mm2): same value with allobjectives

CONDENSER

Microscope stage

The light source, in our case an LED (1), emits

light which is redirected on the sample through

the objective by means of a filter set (2) mounted

on the filter slider.

Fluorescence emitted by the sample is collected

by the same objective (3).


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In reflected light fluorescence the sample is illuminated by the objective which acts as a real

condenser. The capacity of an objective to concentrate light is depending by its numerical aperture,

which typically increases with magnification factor. Therefore illumination provided by a 40x

objective is more efficient than a 10x.

FluoLED for reflected light fluorescence is very we ll indicated for those applications where

medium to high power objectives are used, typically from 20x up to 100x.

Very well indicated also for UV excitation.

Power density (W/mm2): increased by numerical aperture


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Sales arguments

This section explains the main sales arguments of FluoLED. Please note that many of these are

common to both transmitted and reflected light models.

LED modules are light sources emitting an extremely efficient spectrum only in the desired

bandwidth, thus ensuring a very good signal-to-noise ratio.

They are available from near UV across the visible light spectrum. Standard colors are: 630nm

(red), 590 nm (yellow), 535nm (green), 480nm (blue), 450nm (deep blue), and 365nm (UV).

Key Product Benefits:

• One, two and three color excitation. Depending on LED and filter sets, 2- and 3-colour fluorescence observations are possible. Each color channel is dimmable with variable light control, thus allowing to reduce more intense signals in respect to weaker ones.

• No alignment of light source. The LED cassette fits into the adapter without use of any tool.

No risk of low fluorescence signals due to light source misalignment.

• Light source lifetime > 30.000 hrs. Corresponds to 15 years, considering a use of 8 hours a day for 5 days a week. Mercury lamps have an average lifetime of 150-200 hours.

• Regulation of each single color channel with variable light control. Allows to reduce

photobleaching.

• No warm-up time. LED source reaches desired efficiency immediately after turning power on.

• High S/N ratio. LED source emits energy only in the desired wavelengths range, thus

allowing transmitted light fluorescence. • Fluorescence unit is added to the microscope between stand and observation tube,

allowing the use of the original transmitted light illumination system through free position in filter slider.

• Battery pack option for operation on field.

FRAEN AFTER® cassettes Excitation

UV 365 nm

ROYAL BLUE 450 nm

BLUE 480 nm

CYAN (on request) 505 nm

GREEN 535 nm

YELLOW 590 nm

RED 630 nm


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Specifications

List of available LEDs for fluorescence excitation

UV (Dapi)………………………………………………………….………………..……excitation λ 365 nm

Royal Blue (Auramine O, Acridine Orange)…………………......................……… excitation λ 450 nm

Blue (FITC, Alexa 488, EGFP)…………………..………………………….……….. excitation λ 480 nm

Green (Cy3, Alexa 532, Rhodamine, TRITC...………………………………….….. excitation λ 535 nm

Yellow (Texas Red, Alexa 594)…………………...………………………...……….. excitation λ 590 nm

Red (Cy5, Alexa 633, Alexa 647)………………………………………...………….. excitation λ 630 nm

Cyan available on request……………………………………………………………. excitation λ 505 nm

Microscope compatibility

FluoLED reflected light fluorescence illuminator is available for the following Olympus microscope

models:

CX21, CX31, CX41, BX41, BX51

In principle FluoLED reflected light fluorescence illuminator is compatible with any upright light

microscope with infinite corrected optics (usually this sign “∞∞∞∞”is printed on the objectives), provided

the following conditions are fulfilled:

1) Availability of junction kit. This is consisting of two interfaces (dove tails) which allow to

mount the unit on the stand and accept the observation tube.

2) When using multi colour unit depth must be adequate to dimension of microscope stand.

Therfore please inquire before quoting for microscopes which do not belong to the list above.

Transport conditions

Temperature range (with original packaging)...................................-40°C to + 70°C (-40°F t o 158°F)

Storage conditions

Ambient temperature range.................................................................. 5°C to +40 °C (41°F to 104°F)

Operating environment

Operating temperature ......................................................................... 5°C to + 40 °C (41°F to 104°F)

Maximum relative humidity ............................................................................................... 75% at 35°C

Operating air pressure........................................................................................ 800 hPa to 1060 hPa

Altitude...............................................................................................max. 2000 meters (6.561,7 feet)

Pollution degree......................................................................................... 2 (according to IEC 60664)

Use ...................................................................................................................................... Indoor use


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AC:DC power converter

AC:DC power converter ..…………………………………100-240VAC 50/60Hz input, 7.5V dC output Operating temperature ................................5°C to +40 °C (41°F to 104°F), according to UL 60 950-1 Power cord ..................................................................according to IEC 53, length < 2 metres (78,7”) Wire gauge ..............................................................................................................................18 AWG Variable light control for single fluorescence excitation

Input………………...………………………………………………………………...…7.5 V dC, 2 A, 15 W Output………………………………...………………………………………..……..…7 V dC, 1,4 A, 10 W Variable light control for multiple fluorescence excitation

Input………………...……………………………………………………………..….…7.5 V dC, 2 A, 15 W Output………………………………...…………………………………………...….…7 V dC, 1,4 A, 10 W Non-variable light control for single fluorescence excitation

Input………………...………………………………………………….….……………7.5 V dC, 2 A, 15 W Output………………………………...…………………………………………….….…7 V dC, 1 A, 10 W

Documents

FluoLED reflected light fluorescence illuminator - · PDF file• Medical and biological ... FluoLED reflected light fluorescence illuminator is available for the following Olympus