Method for positioning biological samples in a microscopic arrangement

Abstract

The invention relates to methods for positioning at least one preferably biological specimen in the specimen space of a microscope arrangement, and to devices for carrying out these methods. Methods and devices arc proposed, wherein the specimen's orientation relative to a detection objective's optical axis can be repeatedly changed and, in doing so, the specimen is held so that a substantially unobstructed view of the specimen is ensured from every detection direction. In different constructional variants, the specimen is held at a supporting device by adhesive forces or by a flowing medium, the specimen is held at a capillary opening by capillary action, or at least one specimen is embedded in a body of transparent gel, and the gel body is fixed in the specimen space by means of a rotatable holding device, and the detection direction is changed by rotating the holding device by a given angle of rotation.

1 . A method for embedding a specimen in a body of gel, comprising:
 inserting a first partial amount of the gel into a first half of a syringe that is divided in longitudinal direction;   placing the specimen on this partial amount;   placing a second partial amount of the gel on the specimen;   putting the second half of the divided syringe under pressure so that the gel is shaped into a gel body as a result of the cylindrical hollow shape formed between these two halves, the outer diameter of the gel body corresponding to the inner diameter of the syringe; and   enclosing the specimen in the gel body.   
     
     
         2 . A method for centering a specimen embedded in a body of hardening gel inside the gel body, comprising:
 moving the gel body, which comprises gel in a not yet hardened state, into a monitoring station with the enclosed specimen;   evaluating the position of the specimen inside the gel body by means of the monitoring station; and   as a result of the evaluation, orienting the gel body relative to the Earth's gravity in such a way that the specimen drops into the desired position within the gel body under the influence of gravitational force.   
     
     
         3 . A method for positioning a partial amount, as specimen, taken from a biological substance in the specimen space of a microscope arrangement, comprising:
 pressing a syringe with a sharply ground end facing the substance into the substance;   holding the partial amount is held in the syringe while withdrawing the syringe from the substance by means of a plunger which is guided so as to be displaceable inside the syringe and which is connected to a push rod linkage;   moving the syringe with the partial amount into the specimen space, and pushing the partial amount partially out of the syringe by means of the push rod linkage and plunger; and   detecting the pushed out portion as a sample while the remaining portion is held by the syringe.   
     
     
         4 . A device for positioning at least one biological specimen in the specimen space of a microscope arrangement for detecting the specimen, comprising:
 a reservoir for gel bodies of identical shape and size, a specimen being enclosed in each of the gel bodies;   a holding device located in the specimen space of the microscope arrangement for holding one of the respective gel bodies;   a removing and depositing device for removing individual gel bodies from the gel body reservoir and depositing them in the holding device; and   a removing and transporting device for removing and transporting the gel bodies from the holding device.   
     
     
         5 . The device according to  claim 4 , further comprising:
 an image evaluating device and a sorting device which can be controlled depending on the results of the image evaluation and by means of which the individual gel bodies are sorted into different storage vessels after removal from the holding device.   
     
     
         6 . A device for positioning at least one biological specimen in the specimen space of a microscope arrangement for detecting the specimen, comprising:
 a gel body in which a plurality of specimens are enclosed; and   a forward feed device which receives the gel body and which, when actuated, causes the gel body to be displaced in such a way that the enclosed specimens are guided through the specimen space successively in time and, in so doing, a three-dimensional image of each specimen is obtained.   
     
     
         7 . The device according to  claim 6 , further comprising:
 an image evaluating device;   a motor drive for the forward feed device; and   a control circuit which communicates with the image evaluating device and with the drive;   wherein the control circuit is designed in such a way that, after the three-dimensional image is completed, a control command is generated for the drive; and   wherein the control command triggers a forward feed movement and, therefore, causes an exchange of specimens in the specimen space.   
     
     
         8 . A device for positioning at least one biological specimen in the specimen space of a microscope arrangement for detecting the specimen, comprising:
 a gel body having cutouts, each for an individual specimen, at a plurality of positions;   a removing and filling device for removing individual specimens from a specimen reservoir and for inserting them into the cutouts; and   a forward feed device which receives the gel body and which, when actuated, causes the gel body to be displaced in such a way that the cutouts are guided successively in time to an insertion position, where a specimen is inserted into a cutout.   
     
     
         9 . The device according to  claim 8 , further comprising:
 a feed device for an amount of a medium which is initially liquid and which then hardens, this amount corresponding to the shape and size of a cutout, and for inserting this medium into each of the cutouts immediately after the insertion of the specimen.   
     
     
         10 . The device according to  claim 9 , further comprising:
 a heating station and/or cooling station which communicate(s) with the gel body so as to conduct heat and which, before the insertion of the medium into one of the cutouts, causes a heating of the gel at least in the area of the respective cutout and/or, after insertion of the medium into one of the cutouts, causes a cooling of the gel at least in the area of the respective cutout.   
     
     
         11 . The device according to  claim 10 ;
 wherein the forward feed speed and the heating and/or cooling temperature are adapted to the time period required for the hardening of the medium.   
     
     
         12 . The device according to  claim 4  outfitted with a cutting device for cutting out a cutout in the gel body intended for receiving a specimen. 
     
     
         13 . The device according to  claim 12 ;
 wherein a cutting device with a rotating knife is provided for cutting a conical or cylindrical portion of the gel out of the gel body.   
     
     
         14 . The device according to  claim 13 ;
 wherein a syringe is associated with the cutting device for feeding an individual specimen into the cutout.   
     
     
         15 . A device for embedding a specimen in a body of gel, comprising:
 a syringe which is divided into two halves in longitudinal direction for receiving the gel and a specimen; and   a plunger which is guided so as to be displaceable inside the syringe and which is connected to a push rod linkage to push out the gel which is shaped to form a gel body and which, being so shaped, encloses the specimen.   
     
     
         16 . A device for centering a specimen embedded in a body of hardening gel inside the gel body, comprising:
 a syringe which is connected to a rotary drive and in which the gel body of gel with the enclosed specimen is inserted in an as yet unhardened state;   a monitoring station surrounding the syringe, the position of the specimen inside the gel body being evaluated by means of the monitoring station; and   a control circuit for the rotary drive which, as a result of the evaluation, orients the gel body relative to the Earth's gravity in such a way that the specimen drops into the desired position within the gel body under the influence of gravitational force.   
     
     
         17 . A device for positioning a partial amount, as sample, taken from a biological substance in the specimen space of a microscope arrangement, comprising:
 a syringe with a sharply ground end facing the substance; and   a plunger which is guided so as to be displaceable inside the syringe and which is connected to a push rod linkage.