ChemiDoc MP Imaging System - Bio-Rad€¦ · ChemiDoc MP Imaging System Fig. 2. A calibrated...

ChemiDoc™ MP Imaging System

Imaging

Confidence in every step of your imaging workflow.

ChemiDoc MP Imaging SystemBecause results always matter.

Bring a new level of capability and efficiency to your experiments with an imaging system designed for multitasking. The ChemiDoc MP Imager is a unique imaging system that enables stain-free operation and lets you visualize proteins at every stage of your blotting experiment. Its flexibility and sensitivity are complemented by simple, intuitive operation that integrates seamlessly into your workflow.

Bio-Rad Laboratories, Inc.

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Fig. 1. Sensitivity comparison of the ChemiDoc MP System versus X-ray film using blots of serial dilution of transferrin. A, the ChemiDoc Imager delivers superior dynamic range and comparable limit of detection to film. B, a 10-second exposure on film reveals a more limited dynamic range than the ChemiDoc MP System. Saturated pixels are highlighted in red.

Film

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ChemiDoc MP Imaging System

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Sample load, ng

Superior SensitivityGet quantitative, reproducible data without relying on outmoded film processes. The ChemiDoc MP Imaging System offers advanced detection technology that creates optimal exposure for even the faintest bands. Rely on it for fast, super-sensitive chemiluminescence and fluorescence detection and for colorimetric gel and blot documentation.

SUPERIOR SENSITIVITY


Fig. 2. A calibrated luminescent target was used to test light collection efficiency and determine overall system sensitivity. A, data from lower limits of detection are graphed to show overall signal-to-noise ratio. The ChemiDoc MP System delivers top-of-the-class chemiluminescence sensitivity against leading multiplexing imagers on the market. B, images of lower limits of detection from a calibrated luminescent target. The ChemiDoc MP System delivers excellent image quality and limit of detection.

Fig. 2B ChemiDoc MP

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Signal-to-noise ratio of LEDs from a calibrated luminescent test plate. 5 min exposure.


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Fig. 3. 1-D Coomassie-stained gel.

Fig. 4. Chemiluminescent ELISA arrays. Quansys Biosciences Q-Plex Array has 16 distinct capture antibodies bound to each well of a 96-well plate.

Fig. 5. Fluorescent multiplex blot with DyLight 488, DyLight 549, and DyLight 649 conjugates.

Fig. 3

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Exceptional Image QualityWith patented focus calibration technology, images are always in focus at any zoom level. Exceptional dynamic range enables visualization of faint and intense bands on same blot or gel. With Image Lab™ Software you can edit and analyze images on the spot without exporting to other programs.

EXCEPTIONAL IMAGE QUALITY


Cross-talk Cross-talkCorrect Correct

Cross-talk Cross-talkCorrect Correct

FAM A488 Cy3 A546 FAM A488 Cy3 A546

ChemiDoc MP

Fluors detected in green channel Fluors detected in blue channel

Competitor 2

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Fig. 6A. Determination of cross-talk using same multiplex gel. Blue fluors (FAM and Alexa Fluor 488) are detected in the green channel and green fluors (Cy3 and Alexa Fluor 546) in the blue channel using the ChemiDoc MP System and a competitor’s system. For competitor 1, the bold percentage values indicate high cross-talk.

Fig. 6B. Visualization of cross-talk signals recorded with a second competitor’s instrument. The green and blue channel images show high cross-talk values and are indicated in bold. The blue fluors (FAM and Alexa Fluor 488) are detected in the green channel and the green fluors (Cy3 and Alexa Fluor 546) are detected in the blue channel.

Fig. 6B

Fig. 6A

Minimal cross-talk between blue and green channels.


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Fig. 7. Multiplex image file allows multichannel and individual channel view. Image of multiplexed fluorescent blot with Alexa Fluor 488, Alexa Fluor 555, and Alexa Fluor 647.

Unmatched Application VersatilityThe ChemiDoc MP Imager is the only system you need when your experiments include a variety of sample types or require different detection methods. It is the perfect imager to accompany your protein and DNA electrophoresis runs as well as your western blotting experiments. And it delivers quantitative, reproducible results every time.

UNMATCHED APPLICATION VERSATILITY


Fig. 8. Multiple applications of the ChemiDoc MP Imager.

DIGE Criterion™ Gel

Silver-stained 2-D Criterion Gel

Flamingo™-stained Mini-PROTEAN TGX Gel

EtBr-stained wide mini ReadyAgarose™ Gel

Coomassie-stained Mini-PROTEAN® TGX™ Gel

Criterion Stain Free™ TGX Gel

GelRed-stained mini ReadyAgarose Gel

SYPRO Ruby-stained large-format 2-D gel

SYBR® Green-stained wide mini ReadyAgarose Gel


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Fig. 9B. The ChemiDoc MP System is always in focus at any zoom level — no more out-of-focus gel or blot images.

Fig. 9A. The application selected automatically determines the excitation sources and emission filters.

Ease of UseThe ChemiDoc MP System is designed for productivity. With little or no training, users can acquire publication-quality images in seconds. The system is precalibrated to provide the precise focus for any zoom setting or sample height; automated hands-free operation ensures consistent, reproducible, and high-throughput performance.

EASE OF USE


Autofocus

Autoexposure

Autoanalysis

Autoreports

Autooverlay

Autosetup

Autoprint


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V3Visualize

The ChemiDoc MP System will activate and provide immediate visualization of protein separation in all lanes with stain-free gels before blot transfer.

Stain-free gel

Verify

The ChemiDoc MP System paired with stain-free technology enables instant verification of protein transfer before blot detection.

Stain-free blot

Validate

Flexible Image Lab Software tools normalize for protein of interest using total protein stain from stain-free blot and provide quantitative blot results.

Total protein quantification

Protein of interest probed with Alexa Fluor 649

Stain-Free EnabledStain-free technology, offered only by Bio-Rad, eliminates extra steps and unproductive delays in your western blotting experiments.

Stain-free technology is the keystone of the Bio-Rad® V3 Western Workflow™,

a portfolio of products that enables researchers to visualize, verify, and validate results at each step of their western blotting experiment.

STAIN-FREE ENABLED

V3 Western Workflow

Separate Proteins

Run gels in as little as 15 min

■■ Speed with flexibility: TGX Stain-Free™ Gel chemistry available in precast and handcast formats

Efficient and uniform protein transfer in as little as 3 min■■ Throughput: transfer 4 mini gels at once

Quickly assess transfer efficiency■■ Verify quality of transfer for all lanes in 2 min

The V3 Western Workflow streamlines the western blotting protocol, incorporating stain-free in-gel chemistry to allow rapid fluorescent detection of proteins for gels and blots as well as the use of total protein normalization as a loading control. This improved workflow saves time and increases accuracy and reliability throughout the western blotting process.

Transfer

Verify Transfer Efficiency

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Stain-free image of blot

Validate Western Blot Data by Normalization and Analysis

Use stain-free blot image as total protein loading control■■ No need to strip and reprobe■■ Use the entire protein sample in one lane

(no need to rely on housekeeping proteins)■■ Reliable and accurate quantitation

Visualize Protein Separation

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Stain-free image of pretransferred gel

Visualize separation for all lanes in 1 min

■■ Coomassie-like performance with no background variability and no staining/destaining

Workflow

Antibody Incubation and Blot Detection ~5 hr

Benefit

5Detect protein of interest

Normalize protein of interest with stain-free image of blot from step 4



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Total Protein Normalization

MORE RELIABLE to quantitate the protein load1Using total protein normalization produces a much greater linear dynamic range for measuring target protein levels. Housekeeping proteins such as b-actin, b-tubulin, or GAPDH are often very abundant in biological samples, which results in their signal being oversaturated compared to target proteins. Normalizing results to a total protein measurement corrects this problem, allowing a meaningful comparison even with low-abundance targets, and leads to far greater quantitative accuracy in measuring proteins of interest.

Stain-free gel chemistry makes it possible to use total protein levels as a loading control rather than the housekeeping proteins used in traditional western blotting protocols. This negates the need to strip and reprobe the blot and avoids the attendant errors that can be introduced in this step.

2 AVOID FALSE FINDINGS caused by housekeeping protein signal saturation

Stain-free total protein versus housekeeping proteins

Linearity comparison of stain-free total protein measurement and immunodetection of three housekeeping proteins in 10–50 µg of HeLa cell lysate.

HeLa lysate load, µg

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False Findings: Kinase Protein Levels Normalized against Actin

10 40HeLa lysate load, µg

Comparison of kinase protein levels normalized by stain-free total protein or actin loading controls. 10–50 µg of HeLa cell lysate from the same sample were loaded onto a stain-free gel to probe MEK ( ■■), Akt ( ■■), and Erk ( ■■). There should be no changes in the kinase protein levels when data are normalized. Expected value ( ).

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TOTAL PROTEIN NORMALIZATION

Research Papers Using Stain-Free Technology as Loading Controls in Western BlotsMethodology: How stain-free total protein normalization improves western blotting

Stain-free total protein staining is a superior loading control to b-actin for western blots. Gilda JE, Gomes AV (2013). Anal Biochem 440, 186–188.

V3 stain-free workflow for a practical, convenient, and reliable total protein loading control in Western blotting. Posch A, Kohn J, Oh K, Hammond M, Liu N (2013). J Vis Exp, video ID 50948.

Cancer Biology

BRCA2 is epistatic to the RAD51 paralogs in response to DNA damage. Jensen RB, Ozes A, Kim T, Estep A, Kowalczykowski SC (2013). DNA Repair (Amst) 12, 306–311.

Differential network analysis applied to preoperative breast cancer chemotherapy response. Warsow G, Struckmann S, Kerkhoff C, Reimer T, Engel N, Fuellen G (2013). PLoS One, article ID 0081784.

Neurobiology

Proteomic analysis of gliosomes from mouse brain: identification and investigation of glial membrane proteins. Carney KE, Milanese M, van Nierop P, Li KW, Oliet SH, Smit AB, Bonanno G, Verheijen MH (2014). J Proteome Res [published online ahead of print Nov 4, 2014]. Accessed Nov 10, 2014.

Hippocampal extracellular matrix levels and stochasticity in synaptic protein expression increase with age and are associated with age-dependent cognitive decline. Vegh MJ, Rausell A, Loos M, Heldring CM, Jurkowski W, van Nierop P, Paliukovich I, Li KW, Del Sol A, Smit AB, Spijker S, van Kesteren RE (2014). Mol Cell Proteomics 13, 2975–2985.

A trifluoromethyl analogue of celecoxib exerts beneficial effects in neuroinflammation. Penta AD, Chiba A, Alloza I, Wyssenbach A, Yamamura T, Villoslada P, Miyake S, Vandenbroeck K (2013). PLoS One 8, e83119.

Skeletal Muscle

Identification of the immunoproteasome as a novel regulator of skeletal muscle differentiation. Cui Z, Hwang SM, Gomes AV (2014). Mol Cell Biol 34, 96–109.

Immunology

Activation of the P2X7 receptor induces the rapid shedding of CD23 from human and murine B cells. Pupovac A, Geraghty NJ, Watson D, Sluyter R (2014). Immunol Cell Biol [published online ahead of print Aug 26, 2014]. Accessed Nov 10, 2014.

Metabolomics

Quantification of protein copy number in yeast: The NAD+ metabolome. Mei SC and Brenner C (2014). PLoS One 9, e106496.


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Instrument Tour

BlueAutomatic detection of blue fluorescence using dyes such as Cy2, Coomassie Fluor Orange, Alexa Fluor 488, DyLight 488, Pro-Q Emerald 488, Qdot 523, Qdot 605, Qdot 625, Qdot 705.

GreenAutomatic detection of green fluorescence using dyes such as Cy3, Flamingo, Krypton, Pro-Q Diamond, Alexa Fluor 546, DyLight 549, Rhodamine.

RedAutomatic detection of red fluorescence using dyes such as Cy5, Cy5.5, Alexa Fluor 647, Alexa Fluor 680, DyLight 649, DyLight 680, IRDye 680.

Optimized emission and excitation filters for discrete detection of multiplexed visible fluorescence.

INSTRUMENT TOUR


Multiplexed fluorescent imagingMulticolor LEDs optimized for quantitative western blot imaging

Sample tray with UV, white, and blue imaging Flexible transillumination source for easy and uniform imaging of gels and blots

Cooled CCD with autofocus for all zoom levelsUnmatched sensitivity and image quality

6-position automated filter wheel Broad application flexibility

Touch-pad operation For positioning and band excision

Go to bio-rad.com/chemidocmp to see the complete ChemiDoc MP video.

Specifications

Automation Capabilities Workflow automated selection Application driven; user selected or

recalled by a protocol

Workflow automated execution Controlled by a protocol via application-specific setup for image area, illumination source, filter, analysis, and reporting

Workflow reproducibility 100% repeatability via recallable protocols; from image capture to quantitative analysis and reports

Autofocus (patent pending) Precalibrated focus for any zoom setting or sample height

Image flat fielding* Dynamic; precalibrated and optimized for every application

Autoexposure 2 user-defined modes (intense or faint bands)

Hardware specificationsMaximum sample size (L x W) 28 x 36 cm

Maximum image area (L x W) 26 x 35 cm

Maximum image area for 25 x 26 cm standard UV-excited gels (L x W)

Excitation source Trans-UV (302 nm included; 254 nm and 365 nm available as options) and epi-white; optional trans-white conversion screen, XcitaBlue™ Conversion Screen, and epi-red, -green, and -blue LED modules

Illumination control 8 modes available. Trans-UV, epi-white, and no illumination for chemiluminescence are standard; epi-red, epi-green, epi-blue, trans-white, and XcitaBlue Conversion Screens are optional

Detector Supercooled CCD

Image resolution 4 megapixels

Pixel size (H x V) 6.45 x 6.45 µm

Cooling system Peltier

Camera cooling temperature –30°C absolute and regulated

Filter holder 6 positions (5 for filters, 1 without filter for chemiluminescence)

Emission filters 1 included (standard), 4 optional

Dynamic range >4.0 orders of magnitude

Pixel density (gray levels) 65,535

Instrument size (L x W x H) 36 x 60 x 96 cm

Instrument weight 32 kg

Operating RangesOperating voltage 110/115/230 VAC nominal

Operating temperature 10–28°C (21°C recommended)

Operating humidity <70% noncondensing

* U.S. patent 5,951,838.

Ordering InformationCatalog # Description

170-8280 ChemiDoc MP Imaging System with Image Lab Software, PC or Mac, includes darkroom, UV transilluminator, epi-white illumination, camera, power supply, cables, Image Lab Software

Accessories170-8283 Red LED Molecular Kit, pkg of 2 epi-red LED

modules and 1 red emission filter, for use with applications requiring red fluorophore detection

170-8284 Green LED Molecular Kit, pkg of 2 epi-green LED modules and 1 green emission filter, for use with applications requiring green fluorophore detection

170-8285 Blue LED Molecular Kit, pkg of 2 epi-blue LED modules and 1 blue emission filter, for use with applications requiring blue fluorophore detection

170-8289 White Light Conversion Screen, for viewing Coomassie, silver stain, and other colorimetric gel stains

170-8182 XcitaBlue Conversion Screen, includes view goggles; blue conversion screen for viewing SYBR® Green, SYBR® Safe, GFP, Flamingo, and other fluorescent gel stains

170-8183 XcitaBlue Conversion Screen and Filter, includes view goggles and SYBR® Safe Filter (170-8075, 560DF50); blue conversion screen for viewing SYBR® Green, SYBR® Safe, and other fluorescent gel stains

170-8083 Filter 520DF30 62 mm, for SYBR® Green/GFP/SYBR® Gold/fluorescein170-8098 254 nm UV Lamps, pkg of 6170-6887 365 nm UV Lamps, pkg of 6170-8097 Standard 302 nm UV Lamps, pkg of 6170-8089 Mitsubishi Thermal Printer170-7581 Mitsubishi Thermal Printer Paper, 4 rolls170-8184 Gel Alignment Templates, pkg of 3

Software170-9690** Image Lab Software, PC or Mac, for automated

image capture, optimization, and 1-D data analysis

** Included with the imaging system.

Scan this QR code to learn more about the ChemiDoc MP Imaging System, or visit biorad.com/chemidocmp.

Alexa Fluor, Coomassie Fluor Orange, Pro-Q, Qdot, SYBR, and SYPRO are trademarks of Life Technologies Corporation. Coomassie is a trademark of BASF Aktiengesellschaft. Cy is a trademark of GE Healthcare Group Companies. DyLight and Krypton are trademarks of Thermo Fisher Scientific. GelRed is a trademark of Biotium, Inc. IRDye is a trademark of LI-COR Biosciences. Mac is a trademark of Apple Inc. Mitsubishi is a trademark of Mitsubishi Companies. Q-Plex is a trademark of Quansys Biosciences.

Bio-Rad Laboratories, Inc. is licensed by Life Technologies Corporation to sell SYPRO products for research use only, under U.S. patent 5,616,502.

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ChemiDoc MP Imaging System - Bio-Rad€¦ · ChemiDoc MP Imaging System Fig. 2. A calibrated...

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