SpectraMax i3x Multi-Mode Detection Platform

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SpectraMax i3x Multi-Mode

Detection Platform

產品經理楊博甦 Hant Yang

金萬林企業股份有限公司

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• Research

 Microplate reader

 Washer

 Genepix

• Drug Screening

 FLIPR

 Flexstation 3

 ImageXpress

 Ionworks Quattro

About MD products

3

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MD microplate reader—The TOP1 !!

第一台可以動力學偵測第一台連續光譜光度儀 第一台可以Pathcheck®

第一台多功能光度儀

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Imagine a Multi-Mode Plate Reader that evolves over time

3 Mode Base Functionality

Extended Dynamic Range

MiniMax™ Imaging Cytometer option

Improved sensitivity with Spectral Fusion™ Illumination

Ease of Use of SoftMax®

Pro Software

Flexibility of User- Upgradeable Application

Cartridges

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Cartridges – Future Ready

• User-upgradeable within 2 minutes

– No messy upgrades

– No instrument down time – Only cost – new cartridge

• Applications (18 options)

– ScanLater^TM (Western Blot) – AlphaScreen® (PE)

– HTRF® (Cisbio) – Customization – And more

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Each Cartridge Is One Microplate Reader

• Detection Cartridges include:

• Optimal Light Source

– AlphaScreen®: laser diode – FL: LED

• Detection Specific Electronics

• Label Specific Optics

– Excitiation filter (s) – Emission filter (s) – Dichroics

– Polarizer(s) for FP

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Detection Cartridge Options – How they work

LED,

Flash lamp, or Laser diode

Reference Diode

Sample

Channel 1 Channel 2

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HTRF with I3x

Reader EC₅₀* Z’ factor Window (Ratio)

Window (%Delta F) SpectraMax i3 19.3

nM

0.91 8747 3024

* Optimal setting for i3 as determined with Cisbio Certification are 30 µs delay, 400 µs integration, 30 pulses

HTRF cAMP HiRange assay – standard curve Cisbio

requirements

Passing value

i3x

Signal/bkg ≥ 40 106.8

DF% High ≥ 600 939.3

DF% Low ≥ 15 24.4

CV Std 0 < 10 3.8

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ScanLater

^TM

Western Blot Detection

• First western blot detection on a multi- mode microplate reader

• Time-resolved fluorescence (TRF) based assay

– Eu 2nd antibody

– Scan now or up to 30 days later

• Cost saving

– Less expensive than buying an gel imager

• Quantitation

– High sensitivity for fg concentrations

• Time-saving protocols

– Eliminate substrate, darkroom, or exposure time requirements

Fluorescence Intensity

Background Fluorescence

1-Excitation

3-Emission Signals

(Europium labeled)

2

Fluorescence intensity emission measurement

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ScanLater

^TM

Advantages Over Other Methods

E U

Flash lamp

Image or use film

Chemiluminescence Fluorescence Intensity TRF

Sensitivity fg pg fg/pg

Dynamic Range 1-2 logs 4 logs 4 logs

Substrate Yes Not needed Not Needed

Stability Hours Weeks Months

Quantitative Semi/indirect measurement Quantitative/direct measurement Quantitative/direct measurement

Output Film/Imaging Imaging Imaging

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Absorbance with SpectraMax i3 Platform

• Light source: Xenon Flash Lamp

• Detection: Photodiode

• Wavelength selection: Dual Monochromator

• Spectral Range: 230-1000 nm

• Read: top to bottom

• Bandwidth: 4nm

• Performance

• System has less stray light than single Monochromator system

– More sensitive

– Linearity up to 4OD

Absorbance scan of Holmium oxide; 300-600nm in 1nm increments

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Micro-Volume DNA/RNA Quantitation

SpectraDrop Micro-Volume Microplate

• SBS format holder + 2 slide design

• 24 or 64 spot microplate

• 2uL or 4uL configurations

• Sensitivity

• 2ng/uL (dsDNA)

• Easy to clean

• Replaceable slide design

LLD < 2 ng/uL

DNA standards ranged from 50 ng/µL down to 2.4 ng/µL

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Fluorescence with SpectraMax i3 Platform

• Light source: Spectral Fusion™ Illumination

• Detection: PMT

• Wavelength selection: 2x2 Monochromator Design

• Read: top and bottom

• Top read no fibers for Ex and Em

• Bottom: fibers for the Ex and Em

• Bandwidth: Ex 9 or 15 nm, Em 15 or 25 nm

• Performance

• 2x2 Monochromator design in combination with Spectral Fusion™ Illumination allow this system to be

– More sensitive

– Higher dynamic range – Full spectral range

Fluorophore i3 LLD Coumarin 200pM Alexa 430 20pM Fluorescein 1pM Rhodamine 5pM Texas Red 10pM Alexa 633 5pM Alexa 647 3pM Alexa 700 10pM

384 Top, Optimized LLD results

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Unique Spectral Fusion

^TM

Illumination

• Revolutionary light source design

– Xenon flash lamp (for deep UV) – Intensified by LEDs in visible range – More powerful

• Increased sensitivity

– Ultimately increasing the sensitivity of the system across the full spectrum

• Auto LED

• High dynamic range for all conc. on one plate

– No need to reread – No saturation

230 nm 1000 nm

Xenon flash lamp

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Em

Ex

• Advantage:

• Dual monochromators for excitation and/or emission provide best stray light elimination

• Limitation

• Loss of excitation and emission signal (poor light efficiency) can lead to decreased sensitivity

• Solution for I3:

• Better light source with Spectral Fusion Illumination overcomes the efficiency limitations of 2x2 Monochromator Design

• More sensitive than other 2x2 Monochromator Design system

Wavelength Selection: 2x2 Monochromator Design

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Extended Dynamic Range

SpectraMax i3 Base System

Est. dyn. range (decades)

Alexa 430 > 4

Fluorescein > 6

Alexa 700 > 5

Fluorescein Dynamic range (decades):

~6.7

Alexa Fluor 700 Dynamic range (decades):

~5.2

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Luminescence with SpectraMax i3 system

• Detection: Cooled PMT

• Wavelength selection: Dual Monochromator for Em only

• Read: top only

• Bandwidth for Em: 15 or 25 nm

• Performance:

– Low instrument background

– High sensitivity across the visible range (most applications)

Firefly luciferase standard curve ranged from 10 µg/mL down to 1 pg/mL

Dynamic range (decades): ~7

ATP standards ranged from 100 µg down to 10 pg Dynamic range ~7 decades

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Luminescence with SpectraMax i3 system

• In SoftMax pro two options for Emission:

1. Read at all wavelengths

2. Read at specific wavelengths (BRET)

a) Choose the BW

b) Choose up to 4 different wavelengths

c) Choose the Wavelength

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SoftMax Pro Powerful Data Analysis

• Extensive Data Analysis Features

• 3D data visualization

• 19 Curve Fit options

• Plate Cloning feature

• Custom formula writing

• Customizable protocol editing

• Now with image analysis features

• Relevant Help information 1

click away

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The SpectraMax® MiniMax Imaging Cytometer

• A field upgradeable option for the SpectraMax i3 system

• Adds imaging capabilities to the plate reader

• Images in bright-field/transmitted light and fluorescence

• Add functionality for:

• Quick check of your cells

• Cellular resolution assays

“When is the last time you actually looked at your

cells?”

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What advantage does MiniMax offer?

Total intensity

MiniMax

“sees” cells

The Plate reader detector

“sees” RFUs

236 Cells 10 Cells 0 Cells

105.7 RFU 47.7 RFU 45.0 RFU

Easily see your low cell concentration, where in plate reader data might be in the noise

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SpectraMax® MiniMax™ 300 Imaging Cytometer

• Combine cellular imaging and microplate assays

• StainFree™ Technology

– Eliminate staining and maintain cell viability – Cell count

– Confluency

• Multi-channel

– Transmitted, green, red channels

– Toxicity assays

• SoftMax® Pro Software

– Powered by MetaMorph®

– Acquire and analyze

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SoftMax® Pro Software for Imaging Analysis

• Visualization

– Well-by-well data or cell-by-cell data

• Diverse analysis types

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Cell Analysis in SoftMax Pro Software

• Cell Count

• Analysis separates and counts cells based on intensity over background and cell size

• Cell Proliferation/Toxicity

• Analysis detects cell coverage in the well- total cell area

• Marker Expression

• Analysis determines level of signal intensity corresponding to protein or marker expression

Other output parameters can be also

measured

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MiniMax Hardware Details

• Single 4X Objective (Nikon PLAN Fluor)

• Proprietary solid state illumination

• Fixed dichroic and filters

– Green 541/108 nm – Red 713/123 nm

• High sensitivity CCD camera (1.25Mpix, 12-bit)

• Custom laser autofocus

• Transmitted Light/Bright-field illumination

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ELISA

SNP genotyping Molecular beacons

ATP assay

Wellular Resolution

MiniMax Bridging the gap between plate readers and HCS

Large Format Intracellular Detail Micronuclei Neurite Outgrowth Intracellular Trafficking

Cellular Resolution

Cell viability GFP assays FRET assays

IMAP

Cell Counting Cell Migration Whole organism

Live - Dead Cell Cycle Mitotic index

Single Data Point

Sub-Wellular Resolution Sub-Cellular Resolution

Plate Reader MinMax

Imaging Cytometer HCS Instrument

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服務團隊陣容

金萬林企業股份有限公司產品技術應用服務

• 鄒志成博士 MD產品服務時間二年

• 楊博甦碩士 MD產品服務時間五年

• 杜宜碩士 MD產品服務時間三年

• 侯沛庠碩士 MD產品服務時間一年

• 王舒惠碩士 MD產品服務時間一年維修工程師

• 王佑毓 MD產品維修年資七年

• 陳建因 MD產品維修年資三年 Molecular Devices台灣分公司

資深產品應用專員維修工程師

• Samantha Lee ． Jim Chou

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Your success is our success

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Thank you for your attention!

Any Question?

http://www.kimforest.com

http://www.moleculardevices.com Hant@kimforest.com

產品經理楊博甦 Hant Yang 服務電話：0980-580-155 金萬林企業股份有限公司免付費：0800009695

SpectraMax i3x Multi-Mode Detection Platform