Simultaneous false‐colour imaging of birefringence, extinction and transmittance at camera speed

Abstract

SummaryA polarized light imaging technique is introduced that simultaneously captures and unfolds transmittance, fast/slow axes directions and birefringence or linear dichroism from single camera exposures. The technique, based on the rotating polarizer method, is capable of sampling an object through polarizers at four different angles in a single image with the use of an image multiplexer. The range of possible applications is demonstrated with examples from the fields of biology, medicine, forensics and material sciences.

A quantitative collagen fibers orientation assessment using birefringence measurements: Calibration and application to human osteons

“…An automated technique was applied, based on the rotating polarizer method to unfold quantitatively birefringence, eigenmodes and absorption, where an object is simultaneously imaged at four different polarizer angles in a single image implementing an image multiplexer (Quadview, MAG Biosystems) (Kaminsky et al, 2007). In this way the noise contributions coming from camera drifts, fluctuations of light source and mechanical instabilities of the optical system set-up are minimized (Kaminsky et al, 2007).…”

“…Quantitative birefringence assessment requires additional components for the optical set-up (like optical retarders, for the full filters arrangement see Kaminsky et al (2007)). Some methods allow quantitative measurements of two dimensional linear birefringence (Geday et al, 2000).…”

“…An addition of a sample tilting stage provides three dimensional maps of birefringence and eigen rays (Pajdzik and Glazer, 2006). Recently, a technique allowing a simultaneous imaging of birefringence from four quadrants of a single image at camera speed was developed by Kaminsky et al (2007). This method, previously applied to some examples from the fields of biology and materials sciences, was here calibrated for the use on mineralized tissues (mineralized turkey leg tendon – MTLT and osteonal bone).…”

Detecting ordered small molecule drug aggregates in live macrophages: a multi-parameter microscope image data acquisition and analysis strategy

“…Similar to conventional, transmitted light microscopy, polarization microscopy instruments can employ polarized, transmitted light to study the structure of supramolecular aggregates within individual cells [4][5][6][7]. Because of the manner in which polarized light interacts with ordered molecular structures, differences in the organization of molecular components can be detected and quantified based on the fraction of light that is transmitted through the material when it is illuminated with polarized light [8]. At the simplest level, the diattenuation anisotropy of a material can be measured as the maximal difference in light transmittance of a perpendicular pair of planes of polarized light passing through a material, as the material is rotated in the direction perpendicular to both polarized planes of light.…”

Polarized light imaging of birefringence and diattenuation at high resolution and high sensitivity

“…These schemes invariably involve the traditional compensator, which is either rotated under computer control (Glazer et al 1996) or replaced by electro-optical modulators, such as Pockel-cells (Allen et al 1963), Faraday rotators (Kuhn et al 2001), and liquid crystal variable retarders (Laude-Boulesteix et al 2004; Oldenbourg and Mei 1995; Shribak 2011). Alternatively, one can split the imaging beam and simultaneously capture several images, analyzing each for a different polarization state (Kaminsky et al 2007; Shribak et al 2006). These schemes also involve quantitative intensity measurements using electronic light detectors, such as photomultipliers or charge-coupled device (CCD) cameras.…”