Process for carrying out an allergy test, process for determining degranulation in cells, device for carrying out an allergy test and microfluidic chip

Patent metadata

Field	Value
Publication number	US-12078632-B2
Application number	US-201716333404-A
Country	US
Kind code	B2
Filing date	Sep 14, 2017
Priority date	Sep 15, 2016
Publication date	Sep 3, 2024
Grant date	Sep 3, 2024

Abstract

A process for carrying out an allergy test is based on a blood sample being taken. The blood sample is contacted in vitro with at least one allergen. At least one allergic reaction or the absence of the at least one allergic reaction is observed via a microscope ( 11 ) directly and/or optically. To make it possible to carry out an allergy test with a higher validity and/or better accuracy, a position of granules is observed via the microscope ( 11 ). The granules are observed in different planes or horizontal planes.

First claim

The invention claimed is:

1. A process for carrying out an allergy test, the process comprising:
contacting a blood sample with at least one allergen in vitro; and
detecting in vitro at least one allergic reaction or an absence of the at least one allergic reaction directly optically by means of a microscope comprising detecting a position of granules of cells by means of the microscope, wherein the cells comprise immunocytes, wherein the immunocytes of the blood sample are arranged in an at least one reaction chamber of a microfluidic chip, the at least one reaction chamber having an at least partially transparent configuration, wherein the granules are detected in a plurality of planes within the at least one reaction chamber, wherein degranulation of the immunocytes of the blood sample is detected with the microscope in case of the at least one allergic reaction, wherein the position of the granules in the immunocytes, a motion of the granules and/or the degranulation is detected within the at least one reaction chamber for an observation time, the observation time being one of about 1 minute to 10 minutes and about 60 seconds to 300 seconds, wherein one of: each of the plurality of planes is different from each other; and each of the planes is a horizontal plane.

2. The process in accordance with claim 1 , wherein degranulation of immunocytes, granulocytes, basophilic cells, and/or mast cells, of the blood sample is determined with the microscope in case of an allergic reaction, and an absence of degranulation is classified as a non-allergic reaction and a presence of degranulation is classified as an allergic reaction on the basis of the direct optical observation of the immunocytes on contacting with the at least one allergen.

3. The process in accordance with claim 1 , wherein before contacting the blood sample with the at least one allergen, the blood sample is processed to increase a percentage of immunocytes, wherein the percentage of immunocytes in the blood sample is increased to at least 5%, and a percentage of red blood cells and/or of white blood cells different from immunocytes, granulocytes, basophilic cells, basophilic granulocytes and/or mast cells is reduced.

4. The process in accordance with claim 1 , wherein the microfluidic chip is used for contacting the blood sample with the at least one allergen, and an at least one reference chamber of the microfluidic chip has an at least partially transparent configuration.

5. The process in accordance with claim 4 , wherein at least one allergen is guided into the at least one reaction chamber, and the at least one allergen is especially guided both into the at least one reaction chamber containing the immunocytes and into a reference chamber without immunocytes, and the reference chamber is associated with the at least one reaction chamber.

6. The process in accordance with claim 4 , wherein immunocytes of the blood sample are arranged on or in the microfluidic chip by means of a microfluidic cell trap, electrophoresis and/or dielectrophoresis, immunocytes are positioned and/or held by means of electrophoresis and/or dielectrophoresis in at least one reaction chamber, and at least one reference chamber is provided as a chamber free from immunocytes by means of electrophoresis and/or dielectrophoresis.

7. The process in accordance with claim 1 , wherein the position and/or a change in the position of granules of immunocytes are determined directly and/or optically by means of the microscope, and an especially digital, holographic microscopy, shearography and/or a quantitative phase contrast microscopy are carried out with the microscope.

8. The process in accordance with claim 1 , wherein live immunocytes of the blood sample are identified by means of the microscope, the identification of live immunocytes being carried out in an automated manner, and only immunocytes identified as live immunocytes are taken into account during the observation and/or the analysis of a reaction of the immunocytes on contacting the immunocytes with the at least one allergen.

9. The process in accordance with claim 1 , wherein a position of granules in immunocytes, a motion of the granules and/or degranulation are detected over the observation time, the observation time being started with the contacting of the blood sample with the at least one allergen, and a plurality of reaction chambers of a microfluidic chip are observed several times consecutively at predefined time intervals during the observation time.

10. The process in accordance with claim 1 , wherein the observation and/or analysis are carried out in an automated manner, comprising using a digital image recording to detect a reaction of the blood sample upon contacting the blood sample with the at least one allergen and/or using an image processing software to provide and/or analyze recorded images.

11. The process for determining degranulation in cells, especially with a process in accordance with claim 1 , in which a blood sample is taken, and in which the blood sample is contacted in vitro with at least one reaction partner, wherein degranulation or the absence of degranulation is detected directly and/or optically by means of a microscope, the determination of the degranulation being carried out on the basis of an observation and/or measurement of a quantitative phase contrast.

12. The process in accordance with claim 1 , wherein the microscope is configured to generate a holographic image.

13. The process in accordance with claim 12 , wherein the microfluidic chip comprises transparent window surfaces in an area of the at least one reaction chamber on two sides facing away from one another, and a reference chamber, with transparent window surfaces associated with the at least one reaction chamber on two sides facing away from one another, the device further comprising a microfluidic cell trap or electrodes for electrophoretic and/or dielectrophoretic positioning of immunocytes in the at least one reaction chamber, wherein no immunocytes are positioned within the reference chamber or the at least one allergen cannot be directed into the reference chamber.

14. The process in accordance with claim 13 , wherein the at least one reaction chamber and/or a reference chamber have a base or a respective transparent window surface of 100 μm×100 μm on the two sides facing away from one another, and the at least one reaction chamber and/or the reference chamber have a height of less than 1 mm.

15. The process in accordance with claim 14 , wherein:
the microscope comprises a digital, holographic microscope, a shearography microscope and/or a phase contrast microscope;
the microfluidic chip is arranged between a light source and an objective lens device; and
the microscope is connected to a computer for viewing, recording and/or analyzing images and/or data.

16. A process for carrying out an allergy test, the process comprising:
contacting the blood sample with at least one allergen in vitro; and
determining in vitro at least one allergic reaction or an absence of the at least one allergic reaction by monitoring a position of granules of cells in a plurality of planes within an at least one reaction chamber of a microfluidic chip during an observation period via a microscope, each of the planes being different from one another and each of the planes being a horizontal plane, the cells comprising immunocytes, wherein the immunocytes of the blood sample are arranged in the at least one reaction chamber of the microfluidic chip, the at least one reaction chamber having an at least partially transparent configuration, th

Assignees

• Univ Bremen
• Univ Wien Med

Inventors

• Van Den Driesche Sander
• Vellekoop Michael
• Falldorf Claas
• Hafner Christine
• Breiteneder Heimo

Classifications

• G02B21/086

  for transillumination only · CPC title

• B01L3/502761

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title

• G01N2800/24

  Immunology or allergic disorders · CPC title

• G01N33/5076

  involving cell organelles, e.g. Golgi complex, endoplasmic reticulum · CPC title

• G02B21/0004

  specially adapted for specific applications · CPC title