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View additional product information for E-Gel™ Imager Qdot™ 625 Filter - FAQs (4466607)
34 product FAQs found
1) Check that the camera hood or activator key is in place to make sure the base is not deactivated.
2) Light bases will "time out," but the power indicator light may stay illuminated. Turn the base off, then back on.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
Depress the activation button for at least 2 seconds, and the imager will transilluminate for 5 minutes rather than 30 seconds.
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This error can occur if there is other software running in the background, like an Internet Explorer browser, or if there is an antivirus program running that is limiting his ability to access some files.
To fix this error, try to:
- Turn off Internet Explorer or any other program running during the install.
- Inactivate the antivirus program (or temporarily uninstall it), install the E-Gel Imager software, then reactivate or reinstall the antivirus program.
- When the error comes up, select "Ignore" to allow the installation to continue instead of "Retry" or "Abort".
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We do not currently offer software that is compatible with Mac computers. Please install the software on a 32-bit or 64-bit PC running either a Windows XP Professional or Windows 7 Professional operating system.
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For our E-Gel Imager, we recommend using the UV light base with the E-Gel Imager Universal Filter for ethidium bromide gels, and either the blue light base with the E-Gel Imager Universal Filter or the UV light base with E-Gel Imager UV/SYBR Filter (green filter) for SYBR gels.
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Both types of software come with each E-Gel Imager system. GelCapture software is used to control the camera hood during image acquisition and to perform basic manipulations to the image once acquired. The GelQuant Express software is used to analyze images after they have been acquired. This includes estimating the size and relative or absolute mass of bands of interest as well as providing a data report for an entire gel if needed. GelQuant Express software can only be used on a given computer while the activation dongle (HASP key) is inserted into a USB port.
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Here are the specifications:
- Connectivity: USB2.0
- Camera power supply: AC: 100-240V, 50-60 Hz; DC: 7.5 V 2.0 A
- Camera hood dimensions: 35.6 cm (height) x 28.4 cm (length) x 20.3 cm (width)
- Electrical requirements: 100-240 V, 50/60 Hz, 0.6 A
- Temperature: Ambient ±5°C to 40°C
- Base dimensions: 11.9 cm (height) x 30.4 cm (length) x 21.4 cm (width) (this is the entire base)
- Viewing surface dimensions: 15 cm x 12 cm (this is the area where the gel is placed)
- Adaptor specifications: Use only the UL Listed adaptor supplied with the E-Gel Imager Camera Hood (100-240 VAC, 50/60 Hz, 0.6 A)
- Weight: 1 kg
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These E-Gel Imager Filters are alternative filters that can be easily utilized by removing the universal filter tray and sliding in the filter tray of your choosing. We offer an orange E-Gel Imager Universal Filter, a green filter optimal for SYBR Green and SYBR Safe stains and those found in E-Gel EX Agarose Gels, and a red filter for use with our Invitrogen Qdot 625 products.
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Yes, you will need to purchase the E-Gel Imager White-Light Conversion Screen (Cat. No. 4473061), that converts blue light emitted by the blue-light transilluminator or UV light emitted by the UV-light transilluminator to white light. This conversion screen is compatible with multiple protein stains including SimplyBlue SafeStain, SilverQuest silver stain, and Coomassie blue stains.
Gels stained with SYPRO Ruby Protein Gel Stain do not need the white-light conversion screen. They can be visualized using a E-Gel Imager Qdot 625 Filter (Cat. No. 4466607).
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
There are three base options for the E-Gel Imager:
1.Blue-light transilluminator base-ideal for innovative DNA stains such as SYBR Safestains, SYBR Green stains, and those found in E-Gel EX Agarose Gels.
2.UV transilluminator base-best for gels traditionally stained with ethidium bromide.
3.E-Gel adapter base-a one-of-a-kind, real-time gel documentation base that is compatible with the E-Gel Go! System or E-Gel iBase/E-Gel Safe Imager System.
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Please use this link (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/nucleic-acid-gel-electrophoresis/e-gel-electrophoresis-system/e-gel-imager-system.html#specs) to view the E-Gel Imager specifications.
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Spinning your ITK Qdot nanocrystals at approximately 3,000 rpm for 3-5 minutes should remove the white precipitate from the supernatant. Use the supernatant immediately.
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The precipitate in the organic ITK Qdot nanocrystals occurs with some frequency. The ITK Qdot nanocrystals sometimes include impurities that show as a white precipitate.
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Blinking is an inherent property of quantum dots; in fact, all single-luminescent molecules blink, including organic dyes. The brightness and photostability of Qdot nanocrystals makes the blinking more visibly apparent. Under higher energy excitation, Qdot nanocrystals blink even faster.
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Appropriate mounting media selection is very important to retain the fluorescence of Qdot nanocrystals. In our studies, Qdot nanocrystals work best with the following mountants:
HistoMount medium (Cat No. 00-8030); best for long term archiving
Cytoseal 60 Mountant
Clarion Mountant
Most polyvinyl alcohol-based mountants (limited storage time, less than weeks)
Water-based mountants (limited storage time, less than week)
Up to 50% glycerol (limited storage time, less than week)
Note: We do not recommend using ProLong mounting media with Qdot nanocrystals as it will quench their fluorescence.
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Freezing will cause the product to aggregate. The Qdot nanocrystals cannot be dispersed into solution after aggregation.
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Once your product undergoes aggregation, it cannot be dispersed back into solution. We recommend purchasing a new product.
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You may occasionally observe a small amount of aggregation of the Qdot nanocrystals during proper storage. To remove any aggregates that may have formed prior to use, we recommend centrifuging the vial at 2,000 x g for 1 min. Pipette only the supernatant and avoid the pellet. In our experience, pelleting any aggregates that may have formed typically results in a loss of less than 10% of the product.
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We have not systematically investigated the energy transfer properties of the quantum dots, though the quantum dots may have useful properties as both energy transfer donors and acceptors. We have investigated the fluorescence of Qdot 605 Streptavidin conjugates that are coupled to each other through a bis-biotin linker, and found that the emission intensity of the materials was unperturbed at any concentration of biotin cross-linker. These results suggest that the interparticle quenching of these Qdot conjugates is negligible.
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The Qdot products contain cadmium and selenium (and tellurium, in the larger particles) in an inorganic crystalline form. We can only advise that you dispose of the material in compliance with all applicable local, state, and federal regulations for disposal of these classes of material. For more information on the composition of these materials, consult the Material Safety Data Sheet.
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We have not investigated the toxicity of the Qdot nanocrystals. The materials are provided in a solution which is approximately 2 mM total Cd concentration. We have demonstrated the utility of these materials in a variety of live-cell in vitro labeling experiments, but do not have systematic data investigating the toxicity of the materials to humans, to animals, or to cells in culture.
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The number of molecules conjugated to one Qdot nanocrystal is based on the ratio of quantum dot:molecule used in the conjugation, the number of available binding sites on the Qdot nanocrystal, and the size of both the Qdot nanocrystal and the molecule of interest. In general, there are 2-3 antibodies, 4-5 biotin molecules, and 6-8 streptavidin molecules per Qdot nanocrystal.
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ITK Qdot nanocrystals use the original formulation of outer polymer provided in the first generation of the Qdot products; except for the Amine-PEG products, the outer polymer does not include PEG. The outer polymer of the standard Qdot nanocrystals includes PEG.
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There are approximately 80-100 functional groups of each Qdot ITK nanocrystal. We use a type of immunosorbent assay to determine the EC50 of each conjugate.
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Yes, you can visualize Qdot nanocrystals using a standard filter; they will excite at any wavelength below their emission. Keep in mind that the lower the excitation value the brighter the Qdot nanocrystal fluorescence output.
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Qdot nanocrystals do not require the use of antifades as they do not photobleach or fade in the same manner as a chemical dye. In our studies, Qdot nanocrystals work best with the following mountants:
- HistoMount medium (Cat No. 00-8030); best for long-term archiving
- Cytoseal 60 Mountant
- Clarion Mountant
- Most polyvinyl alcohol-based mountants (limited storage time, less than a week)
- Water-based mountants (limited storage time, less than a week)
- Up to 50% glycerol (limited storage time, less than a week)
Note: We do not recommend using ProLong or SlowFade mounting media with Qdot nanocrystals.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Hydrophilic Qdot nanocrystals are stored and shipped in borate buffer pH 8.3-9.0, and organic Qdot nanocrystals are stored and shipped in decane.
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When stored at 4 degrees C, Qdot nanocrystals are stable for approximately 6 months. Qdot nanocrystals should never be frozen due to the possibility of aggregation. The temperature stability of Qdot nanocrystals is summarized below. Please note that fluorescence is not temperature dependent.
<0 degrees C: NEVER freeze Qdot nanocrystals - polymer induces aggregation at freezing temperatures.
>4 degrees C: Core/Shell/Polymer stable at 4 degrees C for ~ 6 months. May be filter sterilized using uncharged filters.
<60 degrees C: Core/Shell/Polymer stable at 60 degrees C (as in in situ hybridization).
<65 degrees C: Core/Shell/Polymer stable at 65 degrees C for only ~1 hour, beyond 1 hour, emission drops off.
<100 degrees C: Core/Shell/Polymer stable up to 100 degrees C brief exposure. OK for 5 minutes at 100 degrees C.
<360 degrees C: Only Core/Shell stable up to 360 degrees C.
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Qdot nanocrystals are most stable at pH 6-9, and marginal stability of Qdot nanocrystals is shown down to a pH 5. Qdot nanocrystals should not be used at pH > 9 due to the possibility of self-aggregation and clumping, and Qdot nanocrystals should not be used pH less than 4 as the polymer and exposed core/shell will begin to dissociate. For more information on Qdot nanocrystals and recommended pH ranges, see pH Ranges for Qdot Nanocrystals (https://www.thermofisher.com/us/en/home/brands/molecular-probes/key-molecular-probes-products/qdot/qdot-reg--nanocrystal0.html)
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You can use Qdot nanocrystals with FRET applications in two scenarios:
- Qdot nanocrystals as donors with fluorescent dyes as acceptors
- Lanthanide (terbium, europium, etc.) as donors with Qdot nanocrystals as acceptors
Note: You cannot perform FRET experiments using Qdot nanocrystals as both donor and acceptor.
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We offer amino (PEG), carboxyl, and streptavidin-functionalized Qdot Innovator's Tool Kit ITK Nanocrystals for the preparation of custom conjugates of proteins or other biomolecules. Amino (PEG)-derivitized forms can be coupled to isothiocyanates and succinimidyl esters or with native carboxylic acids using water-soluble carbodiimides. Carboxyl-derivitized forms can be coupled to amine groups of proteins and modified oligonucleotides. Streptavidin-derivitized forms can be bound with biotinylated conjugates to form stable labeled complexes.
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Qdot nanocrystals and bioconjugates are ideal for experiments requiring long-term photostability or single-excitation, multicolor analysis. Some example applications include:
- Flow cytometry
- Cell and tissue staining
- Cell tracking
- WesternDot western blotting
- In vivo imaging
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Qdot nanocrystals offer many advantages over traditional fluorescent dyes:
- Qdot nanocrystals have a broad excitation range, and they can be excited by any wavelength below their emission peak. The lower the excitation wavelength, the higher the extinction coefficient and Qdot nanocrystal brightness.
- Multicolor detection using Qdot nanocrystals can be done using a single excitation wavelength.
- Qdot nanocrystals exhibit a large Stokes shift.
- Qdot nanocrystals have a narrow emission band.
- Qdot nanocrystals have excellent photostability compared to traditional fluorescent dyes.
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A Qdot nanocrystal is comprises four basic layers. Listed from inner core to outer shell, these are:
1) Core nanocrystal (CdSe or CdSeTe): Determines the color of the Qdot nanocrystal
2) Inorganic shell (ZnS): Improves brightness and stability of the Qdot nanocrystal
3) Organic/polymer coating: Provides water solubility and/or functional groups for conjugation
4) Biomolecule: Covalently attached to the polymer shell and can include antibodies, streptavidin, receptor ligands, or oligonucleotides.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.