mithras lb 940 specifications and ordering...
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Power Supply 110 - 230 V; 50 / 60 Hz; 240 VA
Regulations CE, UL, CSA
Temperature Storage: 0° - 40° C
Range Operation: 15° - 35° C
Humidity 10 – 85%, not condensating
Dimensions 472 x 494 x 374 mm (W x D x H),
incl. injectors
Weight 52 – 54 kg (depending on configuration)
Measurement Luminescence
Technologies Top-reading Fluorescence
Bottom-reading Fluorescence (option)
Fluorescence Polarisation (option)
Absorbance (option)
AlphaScreen™ (option)
Detection Unit Low-noise photomultiplier tube in single photoncounting mode
Excitation Sources Halogen lamp, 75 W, 340 to 700 nm;
Laser, 680 nm (option)
Excitation Filters 355 nm, 485 nm; 405 nm, 450 nm, 490 nm,
620 nm (with Absorbance option)
Emission Filters 460 nm, 535 nm; 400 nm, 515 nm, 480 nm,
530 nm (with BRET option);
568 nm (with AlphaScreen™ option)
Performance
Luminescence: detection limit <20 amol ATP
Fluorescence: detection limit <20 fmol FITC (top reading)
Absorbance: linear range 0 – 2 OD
Fluorescence
Polarisation: c.v. 4.4 % with 1 nM Fluorescein
AlphaScreen™: detection limit <12.5 ng standard beads
Dynamic Range >6 orders of magnitude
Crosstalk Low crosstalk through crosstalk reduction design:5 x 10-6
Injection Unit Up to 4 injectors (variable volume: 10 – 100 µl),
JET Injection technology
Injection into 384 well plates
Plate Formats All 96 and 384 well microplates with outer dimen-sions: 86.0 x 128.2 (W x L); tolerance in plateheight: 13.9 – 15.1 mm
With plate height adjustment option:
All 6 well to 1536 well microplates with outerdimensions: 86.0 x 128.2 mm (W x L); tolerance inplate height: 8.0 – 22.2 mm
Petri dishes, Terasaki Plates, filters with respectiveadapters
Plate Height Optional: automatic detection and adjust-
Adjustment ment to varying plate heights (8.0 to 22.2 mm)
Temperature Control Optional: +5°C above RT to 37 °C
Includes cooling unit for PMT
Operation Modes Integral measurement 0.1 – 600 sec
Kinetics measurement (total time up to 24 h)
Repeated measurement (total time up to 24 h)
Plate repeats (up to 999)
Delay (up to 600 sec)
Scanning (up to 100 individual measurementpoints)
Dispensing with 4 independently controlled vari-able volume injectors
Shaking (3 modes, variable amplitude and speed)
BRET and BRET2 assays
Dual-Luciferase® Reporter Gene assays
Readit™ parameter file
AlphaScreen™ settings
FRET settings
Ratiometric assays
Stacker Stacker20 for 20 microplates (option)
Stacker100 for 100 microplates (option) Robotic Integration Optional Interface Serial RS232, 9 pin PC Operating System Win 98, Win 2000, Win NT, Win XP PC Requirements Pentium processor, 500 MHz (or better), CD ROM
drive, display 1024x768 (or better), serial port,parallel port
Software Windows® application BERTHOLD TECHNOLOGIES MikroWin 2000
Order Information Order NumberMithras LB 940 incl. MikroWin 2000 Lite 940-38099-10Mithras LB 940 with temperature controlincl. MikroWin 2000 Lite 940-38099-15 Injector 1 pre-position L, 10 – 100 µl 940-37772-11Injector 2 measurement position L, 10 – 100 µl 940-37772-12Injector 3 measurement position L, 10 – 100 µl 940-37772-13Injector 4 measurement position F, 10 – 100 µl 940-37772-14 Waste pump (for injectors 1 to 3) 940-38883 Plate Height adjustment 940-39364 Robot integration module 940-39354
BERTHOLD TECHNOLOGIES GmbH & Co. KGCalmbacher Str. 22 • D-75323 Bad Wildbad, GermanyPhone +49 7081 177-0 • Fax +49 7081 177-100info@BertholdTech.com • www.BertholdTech.com
Mithras LB 940 Specifications and ordering informations
LB 9
40 ·
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003
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d.N
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PR2
Rev
02
DLR, DLReady logo and Readit are trademarks of Promega Corporation. Dual-Luciferase and Stop & Glo areregistered trademarks of Promega Corporation. AlphaScreen and DeepBlueC are trademarks of PerkinElmerInc. Windows is a registered trademark of Microsoft. To run certain applications on this product may requirelicenses from others.
PMT cooling unit 940-39337Bottom-reading Fluorescence 940-39348 BRET package 940-39350 AlphaScreen™ package 940-39359 Absorbance option 940-39360 Fluorescence Polarisation option 940-39351 Stacker20 940-40056 Stacker100 940-40057 MikroWin 2000 Advanced I 37854-003 MikroWin 2000 Advanced II 37854-006
A Multilabel ReaderBeyond Expectations
Mithras LB 940
BIO
AN
AL
YT
ICA
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NS
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UM
EN
TS
Today’s needs to perform Many different assaysWith Multiple labels
Are now met by One readerWith No compromise in terms of sensitivity.
The Mithras LB 940 operates with dedicated optical paths to ensure thebest available performance for each technology. In consequence, theimplemented detection technologies can be read with the performanceof dedicated instruments. Those consist of luminescence, fluorescence,absorbance and in addition the homogeneous methods BRET, FRET,AlphaScreen™ as well as Fluorescence Polarisation.
Ancient legend ...... says that on December 25th, Mithras was conceived in the
shade of a sacred tree beside a sacred stream, holding a knifeand a torch. A raven from the sun god Sol then gave him a
message that told him to slay the mysterious white bull.Mithras obeyed and tracked down the bull, accompanied by two minor deities, Cautes and Cautophes. When he finally
met up with the mystic bull, he did slay it, using the knife hewas born with. The white bull then became the moon and
Mithras’ vicorious cape became the sky. Supposedly, this evolved into the alteration of night and day. The legend endwith Mithras’ climbing into Sol’s golden chariot and flashing
across the sky in glory.
• Absorbance• AlphaScreen™• Fluorescence Polarisation• Fluorescence intensity• FRET capability• Dual excitation, dual emission
The highlight of any laboratoryMithras LB 940
• Bottom reading fluorescence• Optimised BRET/BRET2 reading• Flash luminescence• Glow luminescence• Multi-wavelength luminescence• Dual luminescence
• Shaking• Scanning• Small footprint• Dynamic Range adaption (DRA)• Temperature control• Robot accessibility• Stackers• 21CRF part 11 compliancy
detect and identify
• Reading of 1 to 1536 well plates• Fast kinetics• Slow kinetics• Live kinetic monitoring• Up to 4 reagent injectors• Dispensing in 6 to 384 well
plates• Fast filter change enabling ratio-
metric assays
The highlight of any laboratoryMithras LB 940
2
Optics
The proprietary Dedicated Optical Path System
(DOPS) guarantees non-compromised perform-
ance for the individual measurement technolo-
gies luminescence, top reading fluorescence,
bottom reading fluorescence, fluorescence
polarisation, absorbance and AlphaScreen™. As
each technology has its own demands on the
optical system only separated light paths
ensure that the needs for high sensitivity and
wide dynamic range in each technology are
met equally.
Excitation & Emission Filters
Software-driven filtercarriers provide quick
change of emission and excitation filters within
one reading sequence especially important for all
applications with a ratiometric readout, e.g.
Indo-1 and Fura 2. If the measurement requires
several wavelength measurements this is possible
with the flexible protocol definition software.
The changing time for adjacent emission filters is
less than 130 ms providing rapid ratio kinetic
measurements.
Selected high quality interference filters are
mounted in the required positions and additional
filters can easily be added by the user if required.
4
3
Plate Formats
Any kind of microplate regardless of its colour
and format can be measured. All microplate
formats from 6 wells up to 1536 wells are sup-
ported. Additionally Petri dishes and Terasaki
plates can be loaded with adapters. There is no
need for any mechanical adjustment – just
select the appropriate plate type in the operat-
ing software.
The plate height adjustment module detects
the height of the plate used and automatically
adjusts the measurement optics for it.
Even filter mats can be handled in the instru-
ment. A specific filter adapter has been devel-
oped for dot blots especially taking care for
low cross talk while maintaining high counting
efficiency.
5
Reagent Dispensing
Up to 4 independently controlled injectors with
variable volume give entire freedom in the selec-
tion of assay sequence. The injectors are located
in measurement and pre-measurement positions
of the luminescence and the fluorescence path
respectively.
JET injection technology stands for precision and
accuracy – better than 98% – and the only reli-
able way of proper mixing of the added
reagents. Tuneable injection speed covers the dif-
ferent demands of viscosities, cells or filling
heights.
The volume is adjustable from 10 to 100 µl – in
increments of 1 µl. This is perfectly fitting the
requirements for reagent addition into 96 and
384 well plates.
Dispensing can even be performed within a run-
ning kinetic measurement, e.g. to watch the
effects of added agonists / antagonists.
detect and identify
Any type of ratio calcula-
tion, e.g. DLR, or other
mathematical formulas can be linked to every
well individually.
Standard curve fitting and calculation of
unknown samples is available for those users
looking for quantification of their results
(Advanced I and II).
All data can be exported in
EXCEL® or ASCII formats.
Instrument set-
tings, user signa-
tures and file
information are stored with the results.
The password system and the audit trail function
of the Advanced I
and II versions are
important tools
for data security.
The highlight of any laboratoryMithras LB 940
MikroWin 2000
The Windows® based software combines
operation and definition of instrument settings
as well as data reduction and evaluation.
The wells for measure-
ment and injection can be
selected inde-
pendently
Various opera-
tions can be
picked and placed in any order to accommodate
for all different requirements of the assays to be
performed.
Kinetic data
reduction and
graphical display
of the respective
curves help the
user to judge
the results.
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21 CFR part 11 statementSystem access is controlled by log-on routines where unique combinations of user name and password are required to enter
operation levels. Unauthorized access is denied and documented. Electronic signatures are provided by user ID and date/time
stamps in data files. Files can be viewed in electronic and printable form. Audit trails are generated. System has been
developed, tested and documented according to ISO 9001 regulations.
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Stacker & Robotic integration
Specially designed for, but not restricted to the
use in HTS departments of drug discovery com-
panies, the robot access module allows for
uncomplicated integration of the Mithras into
any type of lab automation system. Software
and hardware are easy to integrate into exist-
ing robotic or liquid handling systems.
A barcode reader is available on
request for positive plate identification.
The stacker units provide an of-the-
shelf solution for labs facing moderate
to advanced throughput requirements.
Two pre-configured models are available
Stacker20 to handle up to 20 microplates
Stacker100 to handle up to 100 microplates
Based on the BUTLER plate handling robot the
stackers are characterized by the option of
integrating additional equipment at any time.
The stacker operates in an efficient, reliable
and fast manner. Microplates are securely
gripped due to built-in sensors. There is no
plate loss after power failure. Low noise step-
per motor technology ensures fast and precise
plate handling.
Throughput is even further opti-
mised by the integration of the
unique High Throughput
Transfer & Loading (HTTL) sta-
tion minimising the time loss of
transferring read plates back to the stackers.
Plate stacks are
individually
removable and
can be exchanged
by the user.
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detect and identify
The highlight of any laboratoryMithras LB 940
Intracellular Ca++ levels have become important indicators for the acti-
vation state of ion channels and G-protein coupled receptors as well as
for the phases of apoptosis and cell injury. Though the respective kinetics
and the absolute amounts of the Calcium levels are different for each of
these physiological processes there are common ways for monitoring
them. Luminescent labels like Aequorin as well as fluorescent ones are
versatile and widely used solutions for microplate assays. Fura 2 and
Indo-1 provide a ratiometric readout thereby reducing effects caused by
leaking or bleached dyes or varying assay conditions. OregonGreen
offers a fast single wavelength readout especially suited for high
throughput environments.
These assays can easily and reliably be performed with the use of
Mithras´s variable reagent injectors.
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340 nm360 nm
380 nm
Fluorescence resonance energy transfer (FRET)
Fluorescence Resonance
Energy Transfer (FRET)
has for long been known
to be a smart method for
visualization of molecular
interactions of proteins
and nucleic acids. The discovery and further
development of the Fluorescent Protein family as
well as the Cy dyes has led to an increased use in
microplate assays. Another popular donor/accep-
tor combination is FITC and Rhodamine.
FRET is based on the fact that a donor dye can
transfer a part of this energy and excite an accep-
tor dye. Its emission can be detected as soon as
both dyes are in close proximity.
Applications
GPCRs / 7TM receptorsReporter gene assays
Dual-Luciferase® reporter gene assays (DLR™)Ca++ determination
Protein quantificationATP monitoring
Immunoassays (LIA, FIA)Enzyme kinetics
Readit™ SNP determinationDNA probe assays
Receptor binding studiesCell proliferation
Cell viabilityMembrane potential
Cytokine analysisIon channel studies
Nucleic acid quantificationCellular luminescence / Phagocytosis
Protein-protein interactionsKinase activity
Proteases, e.g. MMPsChemotaxisCytotixicityApoptosis
cAMPNADH
Calcium monitoring
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480 nm
480 nm
480 nm530 nm
detect and identify
Homogeneous functional GPCR assay using BRET detectionRenilla luciferase emits blue
light at 480 nm upon addi-
tion of its substrate
coelenterazine.
Activation of receptor (con-
formation change, attach-
ment of G-protein) due to
binding of
ligand.
β-arrestin / eYFP fusion pro-
tein attaches to receptor
enabling energy transfer
between Renilla luciferase
and the eYFP moiety result-
ing in a rising peak of green
light at about 530 nm.
G-protein coupled receptors, also referred to as
7-transmembrane (7TM) receptors, comprise the
largest and most diverse superfamily of proteins
known. To a minor part only the ligands are
known.
Especially in the field of G-protein coupled recep-
tor research, the BRET technology offers the
opportunity for the establishment of a homoge-
neous and universal functional assay, taking
advantage of the fact that β-arrestin (which is
naturally playing a role in the desensitisation of
the receptors) binds to the intracellular part of
the activated receptor.
BRET is based on the fact that the energy derived
from a Renilla luciferase reaction can be used to
excite a fluorescent protein molecule if the latter
is in close proximity to the luciferase enzyme.
There are several advantages of BRET over other
methods. It is a non-radioactive and homoge-
neous technology. The ratiometric signal mini-
mizes interferences from assay conditions and
keep the time management non critical. There is
no auto-fluorescence coming from compounds or
cell and buffer components as no light source is
required.
Effect of agonist AVP on COS7#41 cells expressing VP2-Rlucand β-arrestin-GFP2
The highlight of any laboratoryMithras LB 940
AlphaScreen™ technology for secondary messenger screening
AlphaScreen™ relies on hydrogel coated Donor
and Acceptor beads providing functional
groups for conjugation to biomolecules. The
beads come in close proximity when the bio-
molecules interact by molecular binding, e.g.
antibody-antigen or receptor-ligand. Laser exci-
tation at 680 nm of a photosensitiser present
on the Donor bead results in the production of
singlet oxygen. The singlet oxygen migrates to
react with chemiluminescers on the Acceptor
bead. The chemiluminescers then activate fluo-
rophores emitting light at 520 - 620 nm.
AlphaScreen™ has proven to be a reliable and
sensitive method in detection assays for second-
ary messengers like cyclic AMP or IP3 activity.
Kinase assays based onFluorescence Polarisation
Fluorescence polarisation (FP) is ideal to meas-
ure the binding of a small molecule to a much
larger one. Unbound fluorescent tracers will
depolarise light during emission resulting in
low polarisation, because the unbound mole-
cules tumble more rapidly than bound tracers.
In contrast, bound fluorescent labelled mole-
cules keep the polarisation orientation as their
rotation is inhibited.
Besides cAMP determination and receptor–lig-
and binding assays a variety of kinase assays
has been established. After the kinase assay has
been performed in its usual way a defined
amount of fluorescent labelled phosphorylated
peptide is added together with the phospho-
specific antibody. The fluorescent tracer com-
petes with the phosphorylated kinase substrate.
Polarisation will decrease inversely proportional
to the kinase activity.
10
11
detect and identify
Readit™ SNP determination
SNPs are the most common type of human vari-
ability promising to play key roles in the deve-
lopment of diseases and the response to thera-
peutic treatments. The Readit™ test is an accu-
rate, reliable and convenient affordable ap-
proach which uses a light emitting luciferase
reaction triggered by ATP. The ATP itself is
derived from pyrophosphorolysis and a subse-
quent phosphorylation of ADP upon matched
probe hybridisation.
DNA probe assaysSeveral diagnostic DNA probe
assays based on acridinium ester
labelled oligo-nucleotides are
commonly used providing the most sensitive
detection and diagnosis of infectious diseases.
Reporter Gene Assays
In basic research of gene regulation as well as
in drug discovery the use of luciferases, ß-galac-
tosidases, ß-glucuronidases and secreted alkali-
ne phosphatases have become a standard tool
offering the highest sensitivity.
Especially the dual luminescence type assays,
e.g. Dual-Luciferase® Reporter Gene Assay,
have become a favourite means as they provide
an internal control for transfection
efficiency or general
expression level.
ATP determination
A detection limit of less than 20 attomoles of ATP
per well makes the Mithras to one of the best
suited microplate instruments for the determina-
tion of cell viability, e.g. in tumor chemo-
sensitivity assays, cell proliferation, antibiotic
susceptibility testing or hygiene monitoring.
Luminescent immunoassays(LIA, ILMA)
By exchanging colorimetric substrates of horse-
radish peroxidase or phosphatases with lumi-
nescent ones an 100-fold increase in sensitivity
can usually be achieved. MikroWin software
with the curve fitting option adds convenient
and extensive data evaluation capabilities to
the superb instrument performance.
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