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Microspectrophotometry in Forensic Science

Forensic Science

  1. Janina Ziȩba-Palus

Published Online: 15 SEP 2006

DOI: 10.1002/9780470027318.a1117

Encyclopedia of Analytical Chemistry

Encyclopedia of Analytical Chemistry

How to Cite

Ziȩba-Palus, J. 2006. Microspectrophotometry in Forensic Science. Encyclopedia of Analytical Chemistry. .

Author Information

  1. Institute of Forensic Research, Cracow, Poland

Publication History

  1. Published Online: 15 SEP 2006


Infrared microspectrophotometry is a combination of two techniques – optical microscopy and infrared (IR) spectrometry – which supply specific information about the composition and structure of a given material. The equipment used is a Fourier transform infrared (FTIR) spectrometer connected on line with an optical microscope.

The microscope allows observation of a sample in white light at relatively high magnification; this allows its morphology and microstructure to be established, and facilitates selection of the area which will subsequently be subjected to IR spectrometric analysis. The microscope also permits use of polarized light in investigations.

The spectrometer enables analysis of a sample with a chosen measuring technique – transmission or reflection – with the aim of determining the chemical composition of the sample. It also supplies information about its microstructure and orientation.

This method is particularly useful in the analysis of trace amounts of various substances secured as material evidence in court cases, and in analysis of the homogeneity of the sample, identification of inclusions and contaminants on the surface, and detection of structural defects.

A specific advantage of this method is its ability to unambiguously photograph and record marked and measured areas of the sample and minimize the process of preparing the sample for analysis.

Furthermore, IR microspectrophotometry gives the rare opportunity of studying small crystals or areas in the oriented materials by using polarized IR radiation, both by the transmission and the reflection technique.

Its fundamental drawback is that the physical nature of the microsample may influence the precision of the photometric measurements and cause distortion of the spectra (artefacts) obtained. Furthermore, because only a small area is studied the heterogeneity of the sample and the amount of contaminants may significantly influence the results of spectrometric measurements.

Microspectrophotometry in the ultraviolet/visible (UV/VIS) range is a combination of photometric measuring techniques and optical microscopy. It allows comparison of the color of very small samples of various materials, such as single fibers, tiny amounts of paint, traces of ink or ball-point pen ink on a forged document in an objective way, independently of the sharpness and quality of the observers's vision. It yields immediate information about spectral differences existing between two samples of similar color, which are indistinguishable by use of the optical microscope. Additionally, using appropriate software to analyze the results of microspectrometric measurements, it enables precise measurement of a color, and – applying the theory of colors – gives a color a defined numerical value; this facilitates communication between experts working with this method. The equipment consists of an optical microscope with a spectrometer for analysis in the visible and ultraviolet range, connected to a microcomputer via an analog-to-digital converter.