Troubleshooting and Things

You Must Know About

Quantum Dots
CD

1. What are Quantum Dots?

Quantum dots (QDs) are nanoparticles of semiconductors, which were theorized in the 1970s and
initially created in the early 1980s. If semiconductor particles are made small enough, quantum effects
come into play, which limits the energies at which electrons and holes can exist in the particles.
Because energy is associated with wavelength or color, this means that the optical properties of the
particle can be finely tuned depending on its size. As artificial nanostructures, by controlling their size,
material, and shape, QDs can be made to emit or absorb specific wavelengths or colors of light or
obtain varied properties.
There are three main types of quantum dots:

・ III-V-semiconductors: made of elements of main group III of the periodic table of the elements (boron,
aluminum, gallium, indium) and main group V (nitrogen, phosphorus, arsenic, antimony, bismuth)

・ II-VI- semiconductors: made of elements of transition metal group II (zinc, cadmium) and main group
VI (oxygen, sulfur, selenium, tellurium)

・ Silicon (Si), the standard material of the semiconductor and chip industry

With a reliable manufacturing technology, Creative Diagnostics is capable of churning out large
quantities of nanocrystals where each batch is produced according to the exact same parameters.
Our DiagNano™ products are now widely used in a wide-ranging number of applications including
catalysis, electronics, photonics, composites, sensing, solar cells, fluorescent biological labels and
optical probes for biological and medical imaging.

Tel: 1-631-624-4882
Email: info@cd-bioparticles.com
2. Basic Structure of DiagNano™ Nanocrystals

Core: determines the color of

the DiagNano™ nanocrystal

Shell: improves brightness and stability

of the DiagNano™ nanocrystal

Coating: provides water solubility and/or

functional groups for conjugation

Biomolecules: covalently attached to polymer shell

and can include immunoglobins,
streptavidin, receptor ligands, or
oligonucleotides

3. Advantages of DiagNano™ Nanocrystals over Traditional

Fluorescent Dyes

・ QDs 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 nanocrystal
brightness.

・ When using QDs, multicolor detection can be completed by using a single excitation wavelength.

・ DiagNano™ nanocrystals exhibit a large Stokes shift.

・ DiagNano™ nanocrystals have a narrow emission band.

・ DiagNano™ nanocrystals possess excellent photostability compared to traditional fluorescent dyes.

4. DiagNano™ Nanocrystal Applications

QDs and bioconjugates are ideal for experiments which need long-term photostability or
single-excitation, multicolor analysis, such as flow cytometry, cell and tissue staining, cell tracking,
western blotting and in vivo imaging.
In addition, we also offer amino, carboxyl, GSH, MPA, etc. functionalized quantum dots for custom
covalent conjugation with proteins, antibodies, and other biomolecules. Quantum dots labeling and
conjugation kits with highly efficient and site-specific labeling and conjugating abilities are available too.

Tel: 1-631-624-4882
Email: info@cd-bioparticles.com
5. pH Range of Stable DiagNano™ Nanocrystals

pH

pH > 9 Not recommended

QDs start to self-aggregate or clump

6 < pH < 9 Most optimal stability is in this pH range

5 < pH < 6 Marginal stability is shown in this range

pH < 4 Not recommended The polymer will dissociate

6. Temperature Range of Stable DiagNano™ Nanocrystals

Temperature

<=0°C Never freeze DiagNano™ Nanocrystals

>=4°C Core/Shell/Polymer stable at 4°C for about 6

months

<=60°C Core/Shell/Polymer stable at 60°C

Core/Shell/Polymer stable at 65°C for only

<=65°C
about 1 hour

Core/Shell/Polymer stable up to 100°C for

<=100°C brief exposure. OK for 5 minutes at 100°C.

<=360°C Only Core/Shell stable up to 360°C

Tel: 1-631-624-4882
Email: info@cd-bioparticles.com
7. Solvents Stability of DiagNano™ Nanocrystals

Chemical Chemical

Acetonitrile Unstable H2O2 Unstable

Alcohols OK up to <10% Heavy Metals Variable

Alkanes Stable MgCl2 Stable

Cacodylate (arsenic-based buffer) Unknown Marine salt (sea salt concentrations) OK

Chloroform Unstable NaCl Stable up to 1 M

DMF Stable NaI Stable up to 1 M

DMSO Stable Paraffin-embedding Stable

5% Tween Stable PBS Buffer Unstable

4% Saponin Stable Phenol Unknown

Triton X-100 Unknown Picric Acid Unknown

NP-40 Unknown Organic solvents Stable

Formaldehyde/Paraformaldehyde Unknown Sodium azide Stable

Formalin Unknown Thimerosal Unknown

Glutaraldehyde Unknown Xylene Stable

Glycerol OK up to 50%

8. Mounting Media for DiagNano™ Nanocrystals

In addition to commercial mounting media, there are several media ideal for short-term storage,
namely most polyvinyl alcohol-based mountants (<weeks), water-based mountants (<week) and
glycerol (up to 50%, <week).

Troubleshooting
1. Why is there a small amount of aggregation of my DiagNano™ product
even though I stored it properly?
You may see a small amount of aggregation of the DiagNano™ product during correct storage. Before
official usage, we suggest centrifuging the vial at 2000 × g for 1 minute to remove the aggregates.
Pipette only the supernatant and avoid the pellet.
2. If my DiagNano™ product is completely aggregated, what should I do?
If that happens, we suggest you order a new product because QDs cannot be dispersed back into
solution. Moreover, since freezing will cause the aggregation, do not freeze your QDs.

Tel: 1-631-624-4882
Email: info@cd-bioparticles.com
3. Why is there a loss of fluorescence after I mounted my samples even
though it was brightly fluorescent before that?
It's important to select an appropriate mounting media to retain the fluorescence of DiagNano™

nanocrystals. As mentioned before, choose commercial media or proper mountant according to your

desired storage time.

4. Is it normal that I found my DiagNano™ product blinking?

All single-luminescent molecules blink (including organic dyes), which is an inherent property of QDs.

The brightness and photostability of DiagNano™ nanocrystals make the blinking more visibly apparent.

Actually, the higher energy excitation, the faster they blink. Try β-mercaptoethanol to reduce blinking.

5. Suggestions for very high background DiagNano™ streptavidin QD

conjugate
• Check if the sample has a high level of endogenous biotin
Some tissues, for example, spleen and kidney sections may contain endogenous biotin, resulting in the

nonspecific signal. Thus, check your sample first and then block the sample using an avidin-biotin

pre-blocking step (BSA or normal animal serum) to decrease nonspecific binding.

• Find grainy staining or clumps of fluorescent material in the background

This is caused by the slight aggregation of BSA. You may spin your samples by a benchtop centrifuge

or a filter to remove microscopic precipitates. In addition, a high level of NaCl or other salts in the

incubation buffer may contribute to this too, so try to reduce the overall salt concentration.

• Optimize the concentration of biotinylated secondary antibodies

Although high levels of biotinylated antibody are required to obtain specific labeling, overly high levels

will lead to nonspecific binding of the antibody to the sample. It's necessary to optimize the level of

biotinylated antibody to achieve specific signal in staining.

• Optimize the concentration of your streptavidin QD conjugate

The level of the final probe should be optimized for each conjugate. Generally, concentrations at or

slightly below saturation should have the optimal signal-to-background ratio, while concentrations

substantially higher than saturation may lead to high background levels.

For more information,

view our website: www.cd-bioparticles.com

Tel: 1-631-624-4882 Email: info@cd-bioparticles.com

Fax: 1-631-938-8221 Address: 45-1 Ramsey Road, Shirley, NY 11967, USA
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