Isolation of cellular material under microscopic visualization

• US 6,251,516 B1
• Filed: 02/04/1998
• Issued: 06/26/2001
• Est. Priority Date: 03/01/1994
• Status: Expired due to Term

• Current Assignee: United States Department Of Health And Human Services
• Original Assignee: United States Department Of Health And Human Services
• Inventors: Krizman, David B., Linehan, W. Marston, Chuaqui, Rodrigo, Emmert-Buck, Michael, Trent, Jeffry M., Peterson, John I., Liotta, Lance A., Smith, Paul D., Bonner, Robert F., Goldstein, Seth R.
• Primary Examiner(s): Cole, Elizabeth M.
• Assistant Examiner(s): RUDDOCK, ULA CORINNA
• Application Number:
  US09/018,596
• Time in Patent Office: 1,238 Days
• Field of Search: 428/346, 428/352
• US Class Current: 428/346
• CPC Class Codes: G01N 1/04 in the solid state, e.g. by cutting
• CPC Class Codes: G01N 1/08 involving an extracting tool, e.g. core bit
• CPC Class Codes: G01N 1/2806 Means for preparing replicas of specimens, e.g. for microscopal analysis
• CPC Class Codes: G01N 1/2813 Producing thin layers of samples on a substrate, e.g. smearing, spinning-on G01N1/30 takes precedence
• CPC Class Codes: G01N 1/286 involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising microtomes G01N1/06; pulverising in general B02C; mixing in general B01F
• CPC Class Codes: G01N 1/31 Apparatus therefor
• CPC Class Codes: G01N 15/1468 with spatial resolution of the texture or inner structure of the particle
• CPC Class Codes: G01N 2001/028 Sampling from a surface, swabbing, vaporising
• CPC Class Codes: G01N 2001/2833 Collecting samples on a sticky, tacky, adhesive surface
• CPC Class Codes: G01N 2001/284 using local activation of adhesive, i.e. Laser Capture Microdissection
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 33/50 Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing measuring or testing processes involving enzymes or microorganisms, compositions or test papers therefor; processes for forming such compositions, condition responsive control in microbiological or enzymological processes C12Q
• CPC Class Codes: G02B 21/32 Micromanipulators structurally combined with microscopes
• CPC Class Codes: Y10T 428/2813 Heat or solvent activated or sealable
• CPC Class Codes: Y10T 428/2839 with release or antistick coating

Abstract

A method of microdissection which involves forming an image field of cells of the tissue sample utilizing a microscope, identifying at least one zone of cells of interest from the image field of cells which at least one zone of cells of interest includes different types of cells than adjacent zones of cells, and extracting the at least one zone of cells of interest from the tissue sample. The extraction is achieved by contacting the tissue sample with a transfer surface that can be selectively activated so that regions thereof adhere to the zone of cells of interest to be extracted. The transfer surface includes a selectively activatable adhesive layer which provides, for example, chemical or electrostatic adherence to the selected regions of the tissue sample. After the transfer surface is activated, the transfer surface and tissue sample are separated. During separation, the zone of cells of interest remains adhered to the transfer surface and is thus separated from the tissue sample, the zone of cells of interest may then be molecularly analyzed.

Claims

1. An activatable film for activated use in capture microdissection where a source emits electromagnetic energy outside of a range of human vision, the activatable film comprising:

a film having a normal non-adherence to a biological sample;

the film optically transparent in the range of human vision for permitting the biological sample to be viewed through the film;

the film activatable upon heating for becoming adhesive at an activated region for adhering to a biological sample at the activated region; and

,

a dye on the film which is optically transparent, the dye coupling to and transducing the electromagnetic energy outside of the range of human vision to heat and activate the film to become adhesive at the activatable region.

2. An activatable film for activated use in capture microdissection according to claim 1 and wherein:

the film includes a region for indicia.

3. An activatable film for activated use in capture microdissection according to claim 1 and wherein:

the film is coated with a release agent to avoid bonding to a biological sample.

4. An activatable film for activated use in capture microdissection according to claim 1 and wherein:

the film includes a support substrate; and

,

an activation substrate.

5. An activatable film for activated use in capture microdissection according to claim 4 and wherein:

the activation substrate is chosen from a group consisting of an emulsion layer, a coated film, a separate impregnated web, thermal sensitive adhesives and waxes, hot glues and sealants, ultraviolet sensitive adhesives, ultraviolet sensitive waxes, ultraviolet sensitive curing optical adhesives, and thermal emulsions.

6. An activatable film for activated use in capture microdissection according to claim 4 and wherein:

the activation substrate is selected from the group consisting of ethylene vinyl acetate (EVA), polyurethanes, polyvinyl acetates, thermal sensitive adhesives, thermal sensitive waxes, thermally-activated hot glues, thermally-activated sealants, ultraviolet sensitive curing optical adhesives, thermal emulsions, acetal, acrylic, bismaleimides, cellulosics, epoxy, fluoroplastics, ketone-based resins, liquid crystal polymers, melamine-formaldehyde, nylon, phenolic, polyamide, polyacrylate, polybenzimidazole, polybutylene, polycarbonate, thermoplastic polyester, liquid crystal polymers, polybutylene terephthalate (PBT), polycyclohexylenedimethylene terephthalate (PCT), polyethylene terephthalate (PET), thermoset polyetherimide polyethylene polyester, branched polyethylene, ethylene acid copolymer, ethylene-ethyl acrylate (EEA), ethylene-methyl acrylate (EMAC), ethylene-vinyl alcohol copolymers (EVOH), high-density polyethylene, HMW-high-density polyethylene, Ionomer, linear low-density polyethylene, linear polyethylene, low-density polyethylene, UHMW polyethylene, very low-density polyethylene, thermoplastic polyimide, thermoset polyimide, polymethylpentene, Polyphenylene oxide, polyphenylene sulfide, PPS, polyphthalamide, polypropylene, polypropylene homopolymer, polypropylene impact copolymers, polypropylene random copolymers, silicones, styrenic resins, ABS, [ACS,] acrylic-styrene-acrylonitrile, expandable polystyrene, polystyrene, impact polystyrene, olefin-modified SAN, polystyrene, styrene-acrylonitrile (SAN) and styrene-butadiene copolymers.

7. An activatable film for activated use in capture microdissection according to claim 4 and wherein:

the support substrate is transparent.

8. An activatable film for activated use in capture microdissection according to claim 4 and wherein:

the support substrate is chosen from the group consisting of a transparent polymer and transparent glass.

9. An activatable film for activated use in capture microdissection according to claim 1 and wherein:

the dye couples to and transduces energy in an infrared range.

10. An activatable film for activated use in capture microdissection according to claim 1 and wherein:

the dye is selected from a group consisting of pytalocyanine dyes, indocyanine dyes, and, naphthanlocyanine dyes.

11. An activatable film for activated use in capture microdissection according to claim 1 and further including:

the film is a thermoplastic film.

12. An activatable film for activated use in capture microdissection according to claim 1 and further including:

a marker added to the film for indicating activation.