Microspectrophotometry is the measure of the spectra of microscopic samples using different wavelengths of

electromagnetic radiation In physics, electromagnetic radiation (EMR) consists of waves of the electromagnetic (EM) field, which propagate through space and carry momentum and electromagnetic radiant energy. It includes radio waves, microwaves, infrared, (visible ...

(e.g.

ultraviolet Ultraviolet (UV) is a form of electromagnetic radiation with wavelength from 10 nm (with a corresponding frequency around 30 PHz) to 400 nm (750 THz), shorter than that of visible light, but longer than X-rays. UV radiati ...

, visible and near

infrared Infrared (IR), sometimes called infrared light, is electromagnetic radiation (EMR) with wavelengths longer than those of visible light. It is therefore invisible to the human eye. IR is generally understood to encompass wavelengths from aroun ...

, etc.) It is accomplished with ''microspectrophotometers'', ''cytospectrophotometers'', ''microfluorometers'', ''Raman microspectrophotometers'', etc. A microspectrophotometer can be configured to measure

transmittance Transmittance of the surface of a material is its effectiveness in transmitting radiant energy. It is the fraction of incident electromagnetic power that is transmitted through a sample, in contrast to the transmission coefficient, which is th ...

absorbance Absorbance is defined as "the logarithm of the ratio of incident to transmitted radiant power through a sample (excluding the effects on cell walls)". Alternatively, for samples which scatter light, absorbance may be defined as "the negative lo ...

reflectance The reflectance of the surface of a material is its effectiveness in Reflection (physics), reflecting radiant energy. It is the fraction of incident electromagnetic power that is reflected at the boundary. Reflectance is a component of the respon ...

, light polarization,

fluorescence Fluorescence is the emission of light by a substance that has absorbed light or other electromagnetic radiation. It is a form of luminescence. In most cases, the emitted light has a longer wavelength, and therefore a lower photon energy, ...

(or other types of

luminescence Luminescence is spontaneous emission of light by a substance not resulting from heat; or "cold light". It is thus a form of cold-body radiation. It can be caused by chemical reactions, electrical energy, subatomic motions or stress on a crysta ...

such as

photoluminescence Photoluminescence (abbreviated as PL) is light emission from any form of matter after the absorption of photons (electromagnetic radiation). It is one of many forms of luminescence (light emission) and is initiated by photoexcitation (i.e. pho ...

) of sample areas less than a micrometer in diameter through a modified optical microscope.

Applications

The main reason to use microspectrophotometry is the ability to measure the optical spectra of samples with a spatial resolution on the micron scale. Optical spectra may be acquired of either microscopic samples or larger samples with a micron-scale spatial resolution. Another reason microspectrophotometry is useful is that measurements are made without destroying the samples. This is important when dealing with stained/unstained

histological Histology, also known as microscopic anatomy or microanatomy, is the branch of biology which studies the microscopic anatomy of biological tissues. Histology is the microscopic counterpart to gross anatomy, which looks at larger structures vis ...

or cytochemical

biological Biology is the scientific study of life. It is a natural science with a broad scope but has several unifying themes that tie it together as a single, coherent field. For instance, all organisms are made up of cells that process hereditary ...

sections, when measuring film thickness in

semi-conductor A semiconductor is a material which has an electrical conductivity value falling between that of a conductor, such as copper, and an insulator, such as glass. Its resistivity falls as its temperature rises; metals behave in the opposite way. ...

integrated circuits, when matching paints and fibers ( forensic science), when studying gems and coal (

geology Geology () is a branch of natural science concerned with Earth and other astronomical objects, the features or rocks of which it is composed, and the processes by which they change over time. Modern geology significantly overlaps all other Ea ...

), and in paint/ink/color analysis in paint chemistry or art-work.

Variations

An advantage of the 'microscope spectrometer' is its ability to use microscope apertures to precisely control the area of sample analysis. Flat capillaries can be used for analyzing small liquid samples, up to about 10 micro-liters in volume. Quartz or mirror-based optics can be used for studying samples from the ultraviolet (UV), down to 200 nm, to the near infrared (NIR) up to 2100 nm. Samples that emit electromagnetic radiation via fluorescence, phosphorescence or photoluminescence when exposed to light, can be quantitatively investigated using a variety of excitation and barrier filters. A variety of observations can be made on samples of interest by using different illumination sources such as halogen, xenon, deuterium and mercury lamps. Plane polarized light can also be used for studying

birefringent Birefringence is the optics, optical property of a material having a refractive index that depends on the Polarization (waves), polarization and propagation direction of light. These optically anisotropic materials are said to be birefringent (or ...

samples.

External links

CRAIC Technologies - The Science of MicrospectroscopyLeica - MICROPHOTOMETRY AND MICROSPECTROPHOTOMETRY
Spectroscopy

Applications

Variations

See also

External links