Laser capture microdissection (LCM) extraction device and device carrier, and method for post-LCM fluid processing

• US 9,103,757 B2
• Filed: 08/13/2010
• Issued: 08/11/2015
• Est. Priority Date: 04/26/2000
• Status: Expired due to Fees

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Baer, Thomas M., Reamey, Robert H., Moravick, Keith E., Head, David F., Richardson, Bruce J., Richardson, Derrick A.
• Primary Examiner(s): LUDLOW, JAN M
• Application Number:
  US12/856,182
• Publication Number:
  US 20110059548A1
• Time in Patent Office: 1,824 Days
• Field of Search: 436/63, 436/174, 436/177, 436/178
• US Class Current: 1/1
• 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 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 1/40 Concentrating samples
• CPC Class Codes: G01N 2001/284 using local activation of adhesive, i.e. Laser Capture Microdissection
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

The present invention generally discloses an extraction system that provides a locale for fluid processing and extraction on a post-microcapture transfer film. The extraction system includes a transfer film carrier and an extraction device forming a reservoir. The extraction system selectively excludes regions of the transfer film from the reservoir to advantageously reduce contamination due to matter adhered to the transfer film by non-specific transfer.

Claims

1. A method for extracting biomolecules from a portion of cells from a cell sample on a carrier, wherein the cell sample includes a plurality of cells, the method comprising:

contacting a stand-off portion of the carrier to a substrate, wherein the stand-off portion is configured to maintain a distance between the substrate and a transfer film, wherein a cell sample is included on the substrate;

transferring a portion of cells selected for analysis from the cell sample included on the substrate to the transfer film, wherein the transfer film is included on the carrier;

mating the carrier to an extraction device, wherein the extraction device is configured to remove biomolecules from the portion of cells selected for analysis, wherein the extraction device comprises;

a carrier-receiving portion comprising a plurality of flanges, wherein the carrier-receiving portion comprises an inner surface, wherein the carrier-receiving portion further includes a landing portion having a landing surface, the landing surface defining an inner opening, and

a conduit interconnected to the carrier-receiving portion, the conduit extending between a first opening on the carrier-receiving portion and a second opening at a second end, wherein the carrier-receiving portion is adapted to receive the carrier and to form a reservoir by the carrier contacting the landing portion, wherein the carrier mating with the carrier-receiving portion closes the inner opening to seal the inner opening to prevent fluid flow through a bottom surface;

deflecting the flanges by passing a at least a portion of the carrier between the flanges so as secure the carrier to the extraction device in a compression-fit engagement;

forming the reservoir with the transfer film by contacting the stand-off portion to the landing surface, wherein the stand-off portion and landing surface are configured so that the stand-off portion is not included in the reservoir;

providing fluid into the reservoir to extract biomolecules from the portion of cells from the transfer film; and

removing the fluid from the reservoir.

2. The method of claim 1 wherein transferring a portion of cells selected for analysis from the cell sample to the transfer film includes transferring the portion of cells to the transfer film by specific transfer microcapture;

and the method further includes disposing the portion of cells that is adhered to the transfer film by specific transfer microcapture within the reservoir.

3. The method of claim 1 wherein transferring the portion of cells selected for analysis to the transfer film includes transferring the portion of cells selected for analysis to the transfer film by non-specific transfer micro-capture;

and

the method further includes substantially excluding matter that is adhered to the transfer film by non-specific transfer microcapture from the reservoir.

4. The method of claim 1 further comprising covering the stand-off portion.

5. The method of claim 2, wherein contacting the stand-off portion of the carrier to the substrate includes a non-specific transfer of matter to the stand-off portion.

6. The method of claim 1, wherein the landing portion reduces the extraction of matter from the stand-off portion of the carrier that contacted the substrate.