JP2000275535A - Phase sample and formation of phase sample - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a phase sample which makes it possible to know the value of a phase difference quantity by joining a 2nd transparent substrate to a hollow formation surface of a 1st hollowed transparent substrate by using an adhesive. SOLUTION: On slide glass 1 made of quartz glass, a hollow 3 of 744 nm in depth is formed and cover glass 2 is adhered onto the slide glass 1 with the hollow 3 by using an adhesive. A pattern of the hollow 3 is in a grating shape which differs in pitch and this pattern is used to investigate the relation between the phase difference quantity and resolution. The adhesive has, for example, a refractive index nd=1.538 on the (d) line of a wavelength of 587.6 nm and is given a phase difference by a refractive index difference from the refractive index nd=1.459 of the quartz glass. The phase difference quantity δis found from the difference in refractive index based upon the depth (t) of the hollow 3. Namely, the phase difference quantity δ is represented as δ=t(1.538-1.459).

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a phase difference sample and a method for preparing the phase difference sample for use in a phase contrast observation apparatus such as a phase contrast microscope capable of observing a colorless and transparent object without staining. Things.

[0002]

2. Description of the Related Art A colorless and transparent object is called a phase object. When light is transmitted or reflected by such a phase object, the wavelength and amplitude of the transmitted light and reflected light do not change, but the phase changes. A phase contrast observation device such as a phase contrast microscope is an optical device that visualizes such a phase change of light. That is, by using a phase contrast observation device such as a phase contrast microscope, it is possible to identify and visualize a portion where the thickness or the refractive index is partially different. Specific examples of the phase object include a biological specimen and a mirror specimen having a step.

[0003]

When a biological specimen is used, phase specimens have individual differences, and are absolutely evaluated (for example, evaluation of the size of a pattern or evaluation of the magnitude of a phase difference).
There is a limit as a sample when doing. Further, it is difficult to measure the absolute amount of an object having a small phase difference, and it is difficult to create a standard sample.

Accordingly, it is an object of the present invention to provide a phase sample from which the value of the phase difference can be known and a method for preparing such a sample.

[0005]

According to the present invention, there is provided a first transparent substrate having a concave surface, and a second transparent substrate which is in close contact with the concave surface.
This is a phase sample characterized by being bonded to a transparent substrate using an adhesive, and a method of producing such a phase sample. Since the depth of the depression formed in the first transparent substrate can be controlled by, for example, the etching time, the depression amount can be determined very accurately. Further, since the refractive index of the adhesive to be bonded is known, the amount of phase difference can be accurately determined. Further, in the case of a reflection-type phase sample, the depression on the surface becomes the phase amount as it is. Therefore, a reflection object such as a metal is vapor-deposited on the surface with the depression to create a phase sample.

[0006]

FIG. 1 is a diagram showing an embodiment of the present invention. 1A is a cross-sectional view of a phase sample in which a broken line portion in FIG. 1B is enlarged, FIG. 1B is a side view of a phase table, and FIG. 1C is a plan view of the phase sample. A recess 744 nm in depth is formed on a slide glass 1 made of quartz glass, and a cover glass 2 is adhered to the slide glass 1 with the recess 3 by an adhesive 3. Figure
The pattern of the depression 3 shown in FIG. 1 (c) has a lattice shape with different pitches, and by using this pattern, the relationship between the amount of phase difference and the resolution can be examined. The adhesive 3 has, for example, a refractive index nd = 1.538 at a d-line having a wavelength of 587.6 nm, and a refractive index nd = quartz glass.
A phase difference is given by a refractive index difference from 1.459. The amount of phase difference δ is obtained from the difference in refractive index, where t is the depth on the slide glass 3. That is, the amount of phase difference δ is δ =
t (1.583-1.459). As a result, FIG.
Shows a λ / 1 at a wavelength of 587.6 nm.
The phase difference is 0. At the time of preparing a phase sample, masking is performed on the slide glass 1 having no recesses with a resist pattern, and the portions having no masking are etched to form the recesses 3. The depth of the depression 3 can be adjusted by controlling the etching time.
In this example, a depth of 744 nm was obtained as described above. The masking described above can be processed to a width of about 0.5 μm, and the amount of phase difference can be freely set to λ / 5, λ / 20, etc. by changing the depth of the depression by controlling the etching time. In the example described above, the depression 3 is formed on the slide glass 1, but the same effect can be obtained by forming the depression on the cover glass 2. Further, the slide glass 1 and the cover glass 2 can be replaced with another transparent material. Furthermore, if various patterns with different amounts of phase difference are prepared instead of the lattice-like pattern of FIG.
The ability of the phase contrast microscope can be evaluated. Examples of various patterns include grid patterns, isolated patterns on circles,
There are Siemens Stars, etc. The reflection-type phase sample can be formed by masking the substrate, forming a recess in a portion other than the mask by etching, and thinly depositing a metal reflective substance thereon. In this case, the substrate does not need to be transparent.

[0007]

As described above, according to the present invention, a fine phase sample can be accurately prepared, and a very important phase sample can be obtained.

[Brief description of the drawings]

FIG. 1 is a view showing an embodiment of the present invention, in which (a) is a partially enlarged view, (b) is a side view, and (c) is a plan view.

[Explanation of Codes] Slide glass 2. Cover glass 3. adhesive

Claims (4)

[Claims] 1. A phase specimen, wherein a second transparent substrate is bonded to a surface of the first transparent substrate provided with the depression by using an adhesive. 2. A method for preparing a phase specimen, comprising forming a depression in a surface of a first transparent substrate and bonding a second transparent substrate to the surface using an adhesive. 3. The method according to claim 2, wherein the recess is formed by masking the surface of the first transparent substrate and then forming a recess in a portion other than the mask by etching. 4. The method according to claim 2, wherein the first transparent substrate and the second transparent substrate are made of quartz glass.