SU1041917A1 - Method and device for measuring micro object thermal oscillation - Google Patents

Abstract

1. Measurement method of thermo-oscil. of microscopic objects, including the registration of light refraction at the boundary between two contacting thermometric bodies with temperature-dependent refractive indices, one of which is the environment, characterized in that in order to expand the range of sizes and measurable micro-objects, refraction the light is recorded on the border of the environment and its wall, the layer adjacent to the micro-object. 2. A device for measuring thermo-oscillations of micro-objects, containing a light source, a microscope with an oblique illumination condenser, a camera and a cuvette for the studied objects, characterized in that in order to provide measurements of thermo-oscillations of subcellular structures, an amplifier of biopotentials and a synchronization unit are added to it. moreover, the output of the amplifiers: tepl biopotentials is connected to the input of the synchronization unit, and the output of the synchronization unit is connected to the source of light. 4 SO YNA

Description

The invention relates to thermometry of optical methods and can be used to measure the rapidly varying temperature of micro-objects, mainly biological, tissue and subcellular, in their own environment. There are known methods for measuring thermoecillations of microsubjects, including the measurement of the temperature-dependent characteristics of the thermometric bodies in contact with the objects under study 1. The device for implementing the known method contains a cuvette for the objects under investigation and a unit for measuring temperature dependent parameters. thermometric bodies. A disadvantage of the known method and device is the impossibility of measuring the characteristics of subcellular structures due to the large mass of thermometric bodies. The closest to the present invention is a method for measuring the thermal oscillations of micro-objects, including detecting the refraction of light at the boundary of two thermocouples in contact with different temperature-dependent refractive indices, one of which is the surrounding micro-object. 2 A device for measuring the thermal oscillations of micro-objects contains light source, microscope with oblique condenser, camera and cuvette for the objects under study. In the implementation of the known method, phenetol and quartz powder suspended in it are used as thermometric bodies. The mixed components have similar refractive indices, moreover, the refractive index of phenetol is thermally dependent. When illuminated with white light, the mixture changes color depending on the temperature in accordance with the change in the refractive index of phenetol. A color change register is measured for thermal oscillations. In the microscopic version, the thermometer body is a single microparticle of quartz, wetted, with phenetril. The disadvantage of this method is that it is impossible to measure its implementation, thermo-oscillation of small objects, such as objects of subcellular structures, due to an increase in heat loss in the contact area of the thermometric bodies with the object under study and, consequently, a decrease in the possible frequency of measured oscillations and a decrease in reliability of measurements. A disadvantage of the known devices is the impossibility of carrying out measurements for subcellular structures with rapidly varying temporally according to a random law thermo-OSCILLATIONS. The purpose of the invention is to expand the range of sizes of the measured micro-objects. This goal is achieved by the fact that according to the method, including the registration of light refraction at the boundary of two thermometric bodies in contact with different temperature dependences of refractive indices, one of which is the surrounding object medium, the refraction of light is recorded at the boundary of the environment and its reception. wall layer adjacent to the microg object. In order to provide measurements of thermo-oscillations of subcellular structures into a device for measuring thermo-oscillations of micro-objects, containing a light source, a microscope with a condenser. oblique illumination, camera and cuvette for the objects under study, additionally introduced a biopotential amplifier and a synchronization unit, the output of the biopotential amplifier connected to the input of the synchronization unit, and the output of the synchronization unit connected to the light source. The drawing shows a diagram of the proposed device. The device contains a pulsed light source 1, a microscope 2 with con, a sensor. 3 oblique illumination, photoKaNijpy 4, cuvette 5 for the objects under study, amplifier b of biopotentials and block 7 of synchronization. The refractive index of the near-wall layer and the environment adjacent to the micro-object, in connection with its other organization, are distinguished from the refractive index of the layer adjacent to the near-wall one, and have a different thermal dependence. Proportional to the temperature of a micro-object, the relative difference between these coefficients can be measured in various ways. One of them is to determine the linear scale change behind the object whose coefficient is measured. The wall layer at the surface of subcellular structures has a thickness sufficient for such measurements with an increase of about 1000 times. In the micrographic version, a fiber of constant diameter should be used as a scale object. Thus, the measurement of thermo-oscillations of micro-objects includes the operations placed {and on the object stage of the dark-field microscope scale-up and thermometry-based objects, photographing them and in the long-term calculation of photograms by the formula

m - JlMj2 1 MO

where is m

IM-diameter scale

fiber and micro-object, correspondingly at the level of the wall layer and the layer / adjacent to it;

M.

proportional to temperature relative to the change of scale due to the near-wall layer of the time sequence M

M. M, ....., M and rd Mr-M,

Mr-M, Mr-Mg, give an idea about the thermal oscillations of a micro-object. .

By using another method for determining the refractive difference of the layers, a scale object can be excluded. , at high magnification, a refraction strip or a Becke strip formed by the near-wall layer can be detected around the micro-object. This requires some defocusing of the microscope. Its position at the micro-object depends on the difference in the refractive indices of the contacting bodies and is a function of temperature if one of these coefficients is thermally dependent. .

Measuring the thermo-oscillations of micro-objects by shifting the refractive band also includes photographing the object after selecting its necessary part. The calculation of the photograms are according to the formula

- Cl) lCj

i-- K

where C

and C

the width of the peripheral part of the refraction band, respectively, at the initial and i-time points;

To the degree of increase of the micro-object;

0

R. displacement of the strip proportional to the increase in the temperature of the micro-object after the i-Toro time interval expires.

5 Time sequence

R, 2 RP presents.

thermo-oscillation of a micro-object.

Example. Measurement of thermal oscillations is performed at the facility

The temperature of which varies over time in a known manner and can be measured in a different way than the one proposed. The object of thermometry is the working thread of platinum.

5 resistance thermometers, the temperature of which increases after a stepwise increase in the electric current flowing through it and is recorded with an oscilloscope. Thread has

A diameter of 30 microns allows for immersion in various media and is placed on the table of a biological microscope from a camera and synchronous shooting. The results of measurements and calculations for platinum in water are given in the table.

The increase in platinum temperature on average, ° C

Number of measurements Scope of measurements, ° С

The offset of the refractive band, microns

Scope of measurements, micron

The correlation coefficient .Me) iyiy is an increase in platinum temperature and a strip shift

0.14O, .270,400,76

65520

0.040.030.030.03

1.42.1 2.52.7

Oh, 60,80,80,8

0,00,850,900,90

The magnitude of the rank correlation between the increase in the temperature of the object and the displacement of the refractive band at its edge indicates a connection between these values. The correlation between them becomes less than optimal than a millisecond after the object temperature changes.

The Kramer – Mises calculated for all observations probability of similarity Р 2 between the shift of the refraction band and the logarithm of the temperature increase does not fall below 0.85.

A device for measuring the thermal oscillations of microobjects, realizing the proposed method, works as follows.

The cuvette 5 for the test objects with the test object is placed on the stage of the biological

microscope 2 ,. having a condenser oblique lighting.

Stimulation of excitement of the object under study and control hardening of the biopotential; carried out by extracellular electrodes.

The biopotential signal from the object under study is fed to the amplifier 6 biopotentials and then to block 7 syn: 4ronisation. The signal from the synchronization unit controls the pulsed light source 1, as a result of which the exposure time is matched to the biopotential.

Thus, the proposed method allows to expand the range of sizes of the measured micro-objects in the direction of their reduction, and the device provides the possibility of measuring thermo-oscillations for subcellular structures.

Claims (2)

1. A method for measuring thermal oscillations of micro objects, including recording light refraction at the boundary of two thermometric bodies in contact with each other with temperature-dependent refractive indices, one of which is. environment, characterized in that, in order to expand the size range of the measured microobjects, light refraction is recorded at the boundary of the environment and its wall layer adjacent to the microobject. 2. A device for measuring the thermal oscillations of microobjects containing a light source, a microscope with a slanting condenser, a photo-. a measure and a cuvette for the studied objects, which is different in that, in order to provide measurements of thermo-oscillations of subcellular structures, a biopotential amplifier and a synchronization unit are added to it, the output of the biopotential amplifier is connected to the input of the synchronization unit, and the output of the synchronization unit is connected to a source Sveta. ''