Phase Microscopy part II
Posted: Thu Mar 12, 2020 10:47 am
In the previous entry about phase contrast, I covered a bit of the history and general principals. It was noted that phase contrast is not one technique but highly variable : really it can be as many as are imagined. Sadly, it has become one thing, with most companies opting to produce a general dark phase system.
In part II, I am covering the general design parameters that when changed or adjusted , alter the type of phase and or the contrast level. Different phase types yield differing results with various types of samples, even different thicknesses of the same sample, so it is clearly evident that the dearth of phase contrast types on the market today, is a detriment to quality microscopy, since phase is one of the least expensive variable contrast tools available. Even different levels of contrast have applications separate from one another. Thus phase contrast is a continuum. For brevity, I will be describing a theoretical system that has an annular design, although phase systems with designs other than annular have existed. Thus, the functional parts of the system are called an annulus or annuli.
A phase system begins with a light source that has conjugate planes at the condenser diaphragm( the phase annulus in the phase condenser body), the focus plane( the object plane) and the diffraction plate( the phase annulus in the objective or above the objective, whichever is the manufacturers method). This is called the conjugate plane or the conjugate area.
Additionally, the diaphragm and diffraction plate are fitted with a complementary area. This is the area of the diaphragm outside of the annulus and the area of the diffraction plate outside of the annulus.
The conditions of the diapragm can be changed to yield different results and the conditions of the diffraction plate can be changed to yield different results.
The diaphragm can be adjusted as to the width and size of the annulus. The width and size of the diaphragm annulus can be adjusted relative to the width and size of the diffraction plate annulus to yield different results or the diffraction plate annulus and diaphragm annulus ratio can remain constant but of different proportions and that will yield different results as well.
The objective diffraction plate can be adjusted as to the degree of absorption in both the conjugate and complementary areas and the degree of wave separation in both the conjugate and complementary areas. Adjusting these parameters, will also yield different results.
Given the high degree of variability available to the design of a phase system, it seems surprising that the industry has defaulted to such a small selection of phase options. It is an area very much open to diy engineering.
Below are 4 representative diffraction plates in cross section, showing the absorptive and diffractive layers. Each displays 3 sections of the conjugate area, which represent the central circle and the two outer sections of the outer ring. The two sections of the annular ring are shown as the two smaller sections. The relative size of the various areas on the plate could of course be of any dimensions, as long as the diaphragm was designed in a complementary fashion.
In the images below the conjugate area is designated 0 and the complementary area 1. Images are from Phase Microscopy by Bennet, Jupnik, Osterburg and Richards.
In part II, I am covering the general design parameters that when changed or adjusted , alter the type of phase and or the contrast level. Different phase types yield differing results with various types of samples, even different thicknesses of the same sample, so it is clearly evident that the dearth of phase contrast types on the market today, is a detriment to quality microscopy, since phase is one of the least expensive variable contrast tools available. Even different levels of contrast have applications separate from one another. Thus phase contrast is a continuum. For brevity, I will be describing a theoretical system that has an annular design, although phase systems with designs other than annular have existed. Thus, the functional parts of the system are called an annulus or annuli.
A phase system begins with a light source that has conjugate planes at the condenser diaphragm( the phase annulus in the phase condenser body), the focus plane( the object plane) and the diffraction plate( the phase annulus in the objective or above the objective, whichever is the manufacturers method). This is called the conjugate plane or the conjugate area.
Additionally, the diaphragm and diffraction plate are fitted with a complementary area. This is the area of the diaphragm outside of the annulus and the area of the diffraction plate outside of the annulus.
The conditions of the diapragm can be changed to yield different results and the conditions of the diffraction plate can be changed to yield different results.
The diaphragm can be adjusted as to the width and size of the annulus. The width and size of the diaphragm annulus can be adjusted relative to the width and size of the diffraction plate annulus to yield different results or the diffraction plate annulus and diaphragm annulus ratio can remain constant but of different proportions and that will yield different results as well.
The objective diffraction plate can be adjusted as to the degree of absorption in both the conjugate and complementary areas and the degree of wave separation in both the conjugate and complementary areas. Adjusting these parameters, will also yield different results.
Given the high degree of variability available to the design of a phase system, it seems surprising that the industry has defaulted to such a small selection of phase options. It is an area very much open to diy engineering.
Below are 4 representative diffraction plates in cross section, showing the absorptive and diffractive layers. Each displays 3 sections of the conjugate area, which represent the central circle and the two outer sections of the outer ring. The two sections of the annular ring are shown as the two smaller sections. The relative size of the various areas on the plate could of course be of any dimensions, as long as the diaphragm was designed in a complementary fashion.
In the images below the conjugate area is designated 0 and the complementary area 1. Images are from Phase Microscopy by Bennet, Jupnik, Osterburg and Richards.