Polarized Light Microscopy


Polarization microscopy may be used for quantitative measurement of optical properties of biological specimens and to reveal dynamic structural details without exogenous stains.

Keywords: optical microscopy; inherent contrast; birefringence; dichroism; polarization

Figure 1.

Schematic diagram of a polarizing microscope (see text).

Figure 2.

Schematic diagram of a DPI microscope. The optional components are used to obtain images of the various forms of anisotropy that characterize a sample that are not due to preferential absorption or scattering. If epi‐illumination is used, fluorescence depolarization studies can be performed.

Figure 3.

Illustration of a DPI microscope. On the left the microscope is placed on a vibration‐damped optical table. Under the table is a Jasco spectropolarimeter used to record circular dichroism spectra of small areas in the sample seen through the microscope. On the right is the rack with the electronics.



Beach DA, Bustamante CJ, Wells KS and Foucar KM (1987) Differential polarisation imaging III. Theory confirmation. Patterns of polymerisation of hemoglobin S in red blood sickle cells. Biophysical Journal 52: 947–954.

CinBo J, Finzi L and Bustamante CJ (1988) Design and application of a computer‐controlled confocal scanning differential polarisation microscope. Review of Scientific Instruments 59: 2399–2408.

Finzi L, CinBo J and Bustamante CJ (1989) Direct observation of large chiral domains in chloroplast thylakoid membranes by differential polarisation microscopy. Proceedings of the National Academy of Sciences of the USA 86: 8748–8752.

Finzi L, Ulibarri L and Bustamante CJ (1991) Differential polarisation imaging. V. Numerical aperture effects and the contribution of preferential scattering and absorption to the circular dichroism images. Biophysical Journal 59: 1183–1193.

Fresnel A (1821) Annales of Chemical Physics 2 17: 167.

Inouè S and Hyde (1957) Studies on depolarization of light at microscope lens surfaces. II The simultaneous realization of high resolution and high sensitivity with the polarising microscope. Journal of Biophysical and Biochemical Cytology 3: 831–838.

Inouè S and Kubota H (1958) Diffraction anomaly in polarising microscopes. Nature 182: 1725–1726.

Kim M, Keller D and Bustamante CJ (1987) Differential polarisation imaging. I. Theory. Biophysical Journal 52: 911–927.

Kim M and Bustamante CJ (1991) Differential polarisation imaging. IV. Images in higher born–approximations. Biophysical Journal 59: 1171–1182.

Livolant F, Mickols W and Maestre M (1988) Differential polarisation microscopy (CD and linear dichroism) of polytene chromosomes and nucleoli from dipteran sarcophaga footpad. Biopolymers 27: 1761–1769.

Oldenbourg R (1996) A new view on polarisation microscopy. Nature 381: 811–812.

Oldenbourg R (1991) Biophysical Journal 60: 629–641.

Shurcliffe W (1962) Polarized Light. Cambridge, MA: Harvard University Press.

Walker RA, Inouè S and Salmon ED (1989) Asymmetric behaviour of severed microtubule ends after ultraviolet‐microbeam irradiation of individual microtubules in vitro. Journal of Cell Biology 108: 931–937.

Further Reading

Ruch F (1956) Birefringence and dichroism of cells and tissues. In: Oster G and Pollister A (eds) Physical Techniques in Biological Research, vol. III, pp. 149–176. New York: Academic Press.

Bennett HS (1950) Methods applicable to the study of both fresh and fixed material. The microscopical investigation of biological materials with polarised light. In: McClung R (ed.) McClung's Handbook of Microscopical Technique, chap. IX, pp. 591–677. New York: Hoeber, P.B., Inc.

Inouè S (1961) Basic design and operation. Design for maximum sensitivity. Clark GL The Encyclopedia of Microscopy, pp. 480–485. New York: Reinhold.

Inouè S and Spring KR (eds) (1997) Video microscopy. The Fundamentals, 2nd edn. New York: Plenum Press.

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How to Cite close
Finzi, Laura, and Dunlap, David D(Apr 2001) Polarized Light Microscopy. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0002994]