Thermo Scientific Confidential
Thermo Scientific NanoDrop™ 3300 Positioning
Versatile, small sample-size fluorometer featuring the following benefits:
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Small sample volume (1-2 ul)
– With assays in microgenomics, proteomics and drug discovery requiring progressively
smaller samples, there is a clear need to measure minute amounts of biomolecules.
The NanoDrop 3300 achieves this by measuring low concentrations in very small
volumes. For example, as few as two picograms or one pg/µl of dsDNA can be
measured using a three microliter PicoGreen assay volume. Sensitivity of Alexa 647 is
five femtomoles or 2.5nM. (This lowers the detection limit of nucleic acids [more than
1000-fold] and dyes commonly used for microarray probes [~100-fold] below that of
the NanoDrop 1000 Spectrophotometer.)
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Broad spectrum analysis (400nm-750nm)
– Allows the measurement of up to four fluorophores in a single sample and
visualization of even more.
– Software utility in FRET (fluorescent resonant energy transfer) analysis in which the
emission of one fluorophore provides the excitation for a second fluorophore.
– Ability to visually inspect the emission spectrum gives more confidence in the final
RFU determination.
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Broad excitation range (365-650nm)
– NanoDrop 3300 enables a broad excitation range without the complexity associated
with high end spectrofluorometers, but rather with the simplicity of low end, less
versatile fluorometers. Both the monochromator (complex and expensive) and filter
changes are effectively eliminated through the unconventional use of a white LED in
addition to filtered UV and blue LED sources. The unconventional use of the broad
spectrum, longer wavelength white LED is enabled by the combination of uniquely
clean optics of the retention system and proprietary signal processing. Simply put,
the software applies a virtual filter to remove the excitation source light from the
emission signal.
– As a point of comparison, sophisticated high end fluorometers or spectrofluorometers
are most commonly found in research laboratories. They utilize a monochromator at
the source (with broad range from a xenon or less powerful halogen lamp) and
variable slit width to enable selection of the excitation wavelength and bandpass.
Alternatively, some high end fluorometers rely on a wide variety of filters to select
amongst common excitation wavelengths.
– For comparison from the other side, low-priced fluorometers frequently use single or
multiple sources, each providing excitation over a narrow range. Common sources
include light emitting diodes (LEDs) in the UV, blue, green or red range as well as
mercury lamps to obtain the common 460nm emission wavelength. Others use filters.
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No cuvettes, capillaries, filters or specialized 96-well fluorescent plates
•
Easy to use
– Preprogrammed methods
– User-defined methods
•
Sample applications
– Micro volume quantitation using common nucleic acid assays (PicoGreen, RiboGreen,
SYBR Green, and Hoechst 33258) and protein assays (Quant-iT Protein).
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Common probe dyes including Cy3 and Cy5, their Alexa Fluor counterparts (Alexa 555
and Alexa 647 respectively), Alexa 488, Alexa 546, and Alexa 568 have been excited
with a single source (white LED).
FRET reactions including:
o FITC – Cy5
o FAM – 3’-Dabcyl
o TET – 3’-Dabcyl
o HEX – 3’-Dabcyl
o Alexa 546 – 3’BHQ-2
o Alexa 594 – 3’BHQ-2
Small instrument size
– 20cm X 15cm X 12 cm
Competitive Products
Low-Priced Fluorometers
• Inexpensive RFU measurements
• Disadvantages
– No spectral analysis
– Limited excitation sources
– Larger sample volumes
– Cuvettes needed
– Some require filters
– Cost of disposables
• Examples
– Turner TBS380 & Modulus
– GE (Amersham) DynaQuant
– BioRad-Versafluor
– Turner Barnsted Quantech
– Farrand Optical A-1
High-Priced Fluorometers
• Full spectrum analysis for versatility
• Disadvantages
– Expensive (purchase, maintenance)
– Large footprints
– Larger sample volumes
– Cuvettes needed
– Many require filters
– Cost of disposables
– Light source stabilization period and longevity
• Examples
– Hitachi F-2500
– Horiba Jobin Yvon FluoroMax-3
– Jasco FP-6200
– Shimadzu RF1501 & RF5301PC
– Varian Cary Eclipse
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Plate Readers
• Fluorescent readings (simple to sophisticated) of microtiter plates for higher
throughput
• Disadvantages
– Relatively expensive instrumentation
– Large sample volume
– Costly disposables (e.g., specialized 96-well fluorescent plates)
– Not conducive for single sample or very limited sample analysis
• Examples
– BioScan Chameleon
– BioTek FLx800
– Perkin Elmer Victor2 D
– Tecan Genios-FL
– Thermo Electron Fluoroskan
– Molecular Devices Gemini EM , XPS