Method and apparatus for attaching, detecting and retrieving a single cell on a surface

Abstract

The present disclosure relates to a method for attaching, detecting and retrieving a single cell on a manipulation medium comprising the steps of: attaching the single cell onto the manipulation medium by applying a carrier liquid; partially drying out the carrier liquid; detecting a property of the single cell by rotating the manipulation medium and scanning a light beam over the manipulation medium; loosening the single cell from the manipulation medium without damaging the single cell by manipulating the single cell with an instrument; and retrieving the single cell from the manipulation medium by aspirating the single cell into a tube, wherein the carrier liquid is dried out to a degree corresponding to that the single cell does not move during the rotation of the manipulation medium and that the single cell can be loosened by the instrument without damaging the single cell. The disclosure further relates to an apparatus for detection and manipulation of one or more object(s),

1 . An apparatus for detection and manipulation of one or more object(s), the apparatus comprising:
 a micromanipulation system comprising a control unit, an arm adapted for holding a tube, optionally adapted for holding the tube via one or more tube clamps, wherein the tube is adapted to move and/or push the object(s), and in which the tube is adapted to be moved in one or more directions following instructions from the control unit,   a microscope having a field of view, said field of view comprising at least one manipulation level where the object(s) is/are situated and the field of view being adapted such that the object(s) and the tube can be in the field of view at the same time.   
     
     
         2 . The apparatus according to  claim 1 , said apparatus being configured for attaching, detecting and retrieving a single cell on a manipulation medium. 
     
     
         3 . The apparatus according to any of  claims 1 - 2 , further comprising a manipulation clamp adapted to hold a manipulation medium at the manipulation level. 
     
     
         4 . The apparatus according to  claim 3 , wherein the manipulation clamp is adapted to rotate the manipulation medium. 
     
     
         5 . The apparatus according to any of  claims 3 - 4 , wherein the manipulation clamp is a slide adapted to translate the manipulation medium. 
     
     
         6 . The apparatus according to any of  claims 3 - 4 , in which the manipulation medium is a glass surface. 
     
     
         7 . The apparatus according to  claim 6 , wherein the glass surface is a glass disc. 
     
     
         8 . The apparatus according to any one of the preceding claims, wherein the object(s) is/are cells, such as blood cells. 
     
     
         9 . The apparatus according to any one of the preceding claims, wherein the object(s) is/are contained in a carrier liquid. 
     
     
         10 . The apparatus according to any one of the preceding claims, wherein the tube is a capillary tube. 
     
     
         11 . The apparatus according to  claims 3 - 10 , wherein the manipulation clamp is adapted to hold an isolation medium, said isolation medium being adapted to hold object(s). 
     
     
         12 . The apparatus according to  claim 11 , wherein the isolation medium is a microscope slide. 
     
     
         13 . The apparatus according to  claim 11 , wherein the isolation medium is a container, such as an Eppendorf tube. 
     
     
         14 . The apparatus according to any one of the preceding claims, wherein the tube is a pipette tube. 
     
     
         15 . The apparatus according to any one of the preceding claims, wherein the tube is adapted to scatter the light at wavelengths between 400 nm and 700 nm. 
     
     
         16 . The apparatus according to any one of the preceding claims, wherein the tube is connected to a pump adapted to aspirate with such a force that a cell will remain in an outermost part of the tube. 
     
     
         17 . The apparatus according to any one of the preceding claims, wherein the tube is adapted to aspirate liquid and/or one or more cells. 
     
     
         18 . The apparatus according to any one of the preceding claims, wherein the pump is adapted to aspirate liquid and/or one or more cells. 
     
     
         19 . The apparatus according to  claim 18 , wherein a proximal end of the tube comprises a cell clamp adapted to hold an individual cell, thereby preventing further liquid to be aspirated. 
     
     
         20 . The apparatus according to any one of  claims 17 - 19 , wherein the liquid is a carrier liquid. 
     
     
         21 . The apparatus according to any one of  claims 1 - 20 , wherein the arm and the tube clamp are adapted to hold the tube at an angle relative to the manipulation level. 
     
     
         22 . The apparatus according to any one of  claims 1 - 20 , wherein the arm and the tube clamp are adapted to hold the tube perpendicular to the manipulation level. 
     
     
         23 . The apparatus according to any one of the preceding claims, wherein the arm and the tube clamp are adapted such that the tube is under the microscope. 
     
     
         24 . The apparatus according to any one of the preceding claims, wherein the directions are defined in a three-dimensional room. 
     
     
         25 . The apparatus according to any one of the preceding claims, wherein the microscope is a fluorescence microscope. 
     
     
         26 . The apparatus according to any one of the preceding claims, wherein the microscope is a light microscope. 
     
     
         27 . The apparatus according to any one of the preceding claims, wherein the microscope comprises a filter adapted to colours of the object(s). 
     
     
         28 . The apparatus according to any one of the preceding claims, wherein the light source is mounted on the arm. 
     
     
         29 . The apparatus according to any one of the preceding claims, wherein the control unit is operated via input from a user interface. 
     
     
         30 . The apparatus according to  claim 29 , wherein the user interface is a mechanical control unit in the form of a keyboard or a joystick. 
     
     
         31 . The apparatus according to  claims 29 - 30 , wherein the user interface is based on a computer-implemented program. 
     
     
         32 . The apparatus according to any one of the preceding claims, wherein the control unit is operated via input from a vision system coupled to the microscope. 
     
     
         33 . The apparatus according to any one of the preceding claims, wherein said direction(s) is/are defined by one or more distances which are one or more micrometres. 
     
     
         34 . The apparatus according to any one of the preceding claims, further comprising a light source adapted to emit light, said light source being adapted to scatter the light in the tube in such a manner that the light from the tube can be seen in the microscope. 
     
     
         35 . A method for preparing a manipulation medium such that one or more object(s) can be detected and manipulated, said method comprising the steps of:
 providing the object(s), said object(s) being contained in a carrier liquid;   spreading the carrier liquid on the manipulation medium;   starting a process of drying the carrier liquid; and   stopping the process of drying the carrying liquid, thereby creating a loose bond between the object(s) and the manipulation medium.   
     
     
         36 . The method according to  claim 35 , wherein the step of spreading the carrier liquid is the process of “smearing”. 
     
     
         37 . The method according to any one of  claims 35 - 36 , further comprising the step of applying a cover glass onto the manipulation medium. 
     
     
         38 . The method according to any one of  claims 35 - 37 , wherein the object(s) is/are detected and manipulated in an apparatus according to any one of  claims 1 - 34 . 
     
     
         39 . A method for detection and manipulation of one or more objects, said method comprising the steps of:
 placing a manipulation medium prepared according to any one of  claims 35 - 381  into an apparatus according to any one of  claims 1 - 34 ;   detecting at least one object by means of the microscope; and   manipulating at least one object by observing the at least one object and the tube in the microscope such that the tube moves and/or pushes the object.   
     
     
         40 . The method according to  claim 39 , wherein the step of detecting at least one object comprises an analysis of the at least one object. 
     
     
         41 . The method according to  claim 40 , wherein the step of manipulating at least one object is carried out by the control unit via input from a vision system coupled to the microscope. 
     
     
         42 . The method according to  claims 39 - 41 , further comprising a step of transferring the object(s) by moving them to an isolation medium by means of the tube. 
     
     
         43 . The method according to  claim 42 , wherein the transfer step is carried out by the control unit via input from a vision system coupled to the microscope.