Laser cell purification system

• US 6,316,234 B1
• Filed: 10/28/1999
• Issued: 11/13/2001
• Est. Priority Date: 09/19/1995
• Status: Expired due to Term

• Current Assignee: Bova, G. Steven
• Original Assignee: Bova, G. Steven
• Inventors: Bova, G. Steven
• Primary Examiner(s): Tate, Christopher R.
• Assistant Examiner(s): WINSTON, RANDALL O
• Application Number:
  US09/429,317
• Time in Patent Office: 747 Days
• Field of Search: 435/325, 435/1.1, 435/173.1, 435/173.7, 435/173.9, 435/284.1, 435/286.2
• US Class Current: 435/173.7
• 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 1/34 Purifying; Cleaning processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: G01N 1/44 Sample treatment involving radiation, e.g. heat
• CPC Class Codes: G01N 2001/284 using local activation of adhesive, i.e. Laser Capture Microdissection
• CPC Class Codes: G01N 2015/1497 Particle shape

Abstract

A process and apparatus for cell purification and ablation is disclosed. The present invention comprises a laser system which directs radiant energy at computer or manually selected individual cells thereby disrupting DNA, RNA and protein structure in those cells. The present invention produces a purified tissue section containing relatively intact DNA, RNA or protein from only the untreated cells. This purified sample is suitable for amplification of material by PCR or other techniques for the analysis of molecular genetic features in the selected cells of interest.

Claims

1. A method for purifying a heterogenous cell population in a tissue section, said method comprising:

(a) supporting a tissue section having a heterogenous population of cells on a tissue support under a microscope with an objective for viewing said tissue section;

(b) scanning said tissue section with a video camera and producing a mosaic of video images representing the tissue section;

(c) displaying said mosaic of video images representing the tissue section;

(d) analyzing each frame of said mosaic of video images based upon target cell selection criteria and selecting target cells of interest based upon said target cell selection criteria; and

(e) automatically and selectively ablating each selected target cell utilizing a radiant energy beam.

2. The method of claim 1 further comprising the step of automatically analyzing each frame of said mosaic of video images based upon target cell selection criteria and automatically selecting target cells of interest based upon said target cell selection criteria.

3. The method of claim 1 further comprising the step of visually analyzing each frame of said mosaic of video images based upon target cell selection criteria and manually selecting target cells of interest based upon said target cell selection criteria.

4. The method of claim 2 further comprising the steps of manually selecting additional target cells of interest or manually deselecting target cells automatically selected based on said target cell selection criteria.

5. The method of claim 1 further comprising the steps of:

(a) changing the magnification of said microscope to a higher magnification; and

(b) repeating steps (a) through (e) in claim 10.

6. The method of claim 1 wherein the step of automatically and selectively ablating each selected target cell produces a tissue section having relatively intact DNA, RNA or protein from only the non-selected cells.

7. A method for laser-treating specific target cells in a tissue section having a heterogenous population of cells, comprising:

(a) supporting said tissue section on a tissue support under a microscope with an objective for viewing said tissue section;

(b) scanning said tissue section with a video camera to produce a mosaic of video images representing said tissue section;

(c) optionally displaying said mosaic of video images;

(d) analyzing said mosaic of video images based upon preselected target cell selection criteria;

(e) selecting said specific target cells based upon predetermined target cell selection criteria; and

(f) laser-treating said selected target cells.

8. A method according to claim 7, wherein said analysis and selection are performed automatically by a computer.