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Home > Measurement Techniques > Fluorescence

Fluorescence

Fluorescence

Custom fluorescence system using an Ocean Optics spectrometer. The customer adapted the unit for underwater use.

First observed in 1560, fluorescence has evolved into a powerful technique that enables entire fields of cutting-edge science and medicine. The only spectroscopic technique capable of resolving single molecules, fluorescence has moved from the lab to applications limited only by imagination.

The field of fluorescence is as diverse as it is beautiful. The sample under study may be liquid or solid, irregularly shaped, or accessible only outside the lab. A well-configured modular spectroscopy system can easily be reconfigured to study a wide range of samples in both the lab and the field.

Although fluorescence is sophisticated, it doesn’t need to be that complex. The beauty of modular spectroscopy is that you can invent as you go, trying different configurations until you find something that works. The tips and tricks below can help you get started. Or give us a call. It may just turn out that our team of application engineers have seen your application before. It’s not unusual, even if your application is.

Advantages

  • Sensitive: Concentrations as low as picomoles and femtomoles can routinely be detected, with even lower concentrations possible.
  • Quantitative: Fluorescence signal is generally proportional to concentration. Fluorescence intensity responds to changes in concentration within picoseconds, making it well-suited for in-situ studies and monitoring fast processes.
  • Safe: Unlike many other techniques for studying biological samples, it is non-destructive to the sample, and has no hazardous byproducts.

Applications

Other Common Applications

  • Nature: analysis of gemstones, minerals, chlorophyll, and crude oil residues
  • Forensics: detection of fingerprints & blood; analysis of fibers and other materials
  • Phosphor thermometry: measurement of temperature using the lifetime or intensity ratios of fluorescence peaks
  • Fundamental studies: use of laser-induced fluorescence to study the electronic structure of molecules and their interactions; concentrations in combustion, plasma, and flow phenomena
  • Biology: detection of molecules, observation of cellular processes, and sorting cells
  • Medical diagnostics: analysis of tissue for presence of cancer, glucose sensors, DNA sequencing, cytometry, and gel electrophoresis

Application Notes:

Technical Note

What Is the Difference Between Quick View Mode and Relative Irradiance Mode?

Scope mode data shows the raw number of counts for each pixel in the array without any processing or correction for spectrometer sensitivity. This is important to remember, because each spectrometer has a different response function that comes from a combination of its individual elements and alignment. That can make scope mode misleading, showing a peak in the right general location, but with a distorted shape and/or center wavelength.

Fluorescence Technical Note

This can be corrected by calibrating against a blackbody light source of known color temperature and working in relative irradiance mode. A tungsten halogen lamp is a convenient standard, and works well for visible and NIR wavelengths. A relative irradiance measurement generates a corrected spectrum of relative intensity as a function of wavelength, scaled from 0 to 1 in arbitrary units.

Is relative irradiance mode always necessary for fluorescence measurements? No. Measurements taken with a single spectrometer are accurate relative to one another, even if the spectral shape is uncorrected. That means you can take the ratio of one fluorescence measurement to another and get an accurate change in the percent signal as a function of wavelength.

Relative irradiance is important when comparing measurements taken by different spectrometers, when determining the spectral shape, or when looking for peak location and shifts.

Application Note:

Featured Products for Fluorescence:

Fluorescence

USB4000-FL Spectrometer is preconfigured for fluorescence measurements from 360 – 1100 nm.
The spectrometer comes with a 200 µm slit and detector collection lens for increased light throughput.
QE Pro-FL Highly sensitive spectrometer specifically suited for low light-level applications such as fluorescence.

What light source to select?
What sampling optics do I need?
How do I detect data information?
How do I determine excitation wavelength?
How do I maximize my signal?
What is the best way to measure turbid or opaque samples?
What is luminescence?
Can I use spectrometer for flow cytometry and fluorescence microscopy?
Fluorescence Fluorescence Fluorescence

These are the basic components to perform a fluorescence measurement using  an Ocean Optics Flame spectrometer, software and sampling accessories. First observed in 1560, fluorescence has evolved into a powerful technique that enables entire fields of cutting-edge science and medicine. The only spectroscopic technique capable of resolving single molecules, fluorescence has moved from the lab to applications limited only by imagination.

Flame Spectrometer

Flame Spectrometer

High Thermal Stability, Interchangeable Slits
FLAME-BUNDLE-FL

FLAME-BUNDLE-FL

Application-Ready System for Fluorescence
JAZ Spectrometer

JAZ Spectrometer

Handheld Spectrometer for UV-Vis Measurements
Spark-VIS

Spark-VIS

Ultra-compact Visible Spectral Sensor
QE Pro (Custom)

QE Pro (Custom)

High-sensitivity Spectrometer for Low Light Level Applications
Maya2000 Pro (Custom)

Maya2000 Pro (Custom)

High Sensitivity Spectrometer
Maya LSL Spectrometer

Maya LSL Spectrometer

Low Stray Light with High Sensitivity
Maya2000 Pro-NIR

Maya2000 Pro-NIR

High-sensitivity Spectrometer for Raman and NIR Applications
USB4000-FL

USB4000-FL

Preconfigured Spectrometer for the Fluorescence Applications and Measurements
QE Pro-FL

QE Pro-FL

High-sensitivity Spectrometer for Fluorescence
Ventana-VIS-NIR

Ventana-VIS-NIR

High-sensitivity Spectrometer for Fluorescence Techniques
HL-2000 Family

HL-2000 Family

Tungsten Halogen Light Sources for the Vis-NIR
DH-mini Light Source

DH-mini Light Source

Compact Deuterium, Halogen Light Source for the UV-Vis-NIR
DH-2000 Family

DH-2000 Family

Deuterium-Halogen Light Sources for the UV-Vis-NIR
PX-2

PX-2

Pulsed Xenon Light Source
HG-1 Calibration Source

HG-1 Calibration Source

Mercury Argon Calibration Source
HPX-2000 Family

HPX-2000 Family

High-powered, Continuous Xenon Light Sources
ecoVis

ecoVis

Compact, Low-Voltage Light Source
KR-1

KR-1

Krypton Calibration Source
LLS Family

LLS Family

High-Performance UV and visible LEDs
D-2000 Family

D-2000 Family

Stable, Deuterium Light Sources for the UV
DH-2000-BAL

DH-2000-BAL

Balanced Deuterium, Halogen Light Source for the UV-Vis-NIR
AR-1

AR-1

Argon Calibration Source
UVTOP®

UVTOP®

Deep UV LED Bulbs
XE-1

XE-1

Xenon Calibration Source
NE-1

NE-1

Neon Calibration Source
Raman Sample Holders

Raman Sample Holders

Accessories for Sampling of Liquids, Powders and More
MCLS

MCLS

Multi-Channel LED Light Source
74-series Collimating Lenses

74-series Collimating Lenses

Single and Achromatic Lenses