Devices and methods for producing microarrays of biological samples

• US 6,902,702 B1
• Filed: 11/27/2000
• Issued: 06/07/2005
• Est. Priority Date: 08/16/2000
• Status: Expired due to Fees

• Current Assignee: University Health Network
• Original Assignee: University Health Network
• Inventors: Winegarden, Neil A., McNeil, James B.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): Nagpaul, Jyoti
• Application Number:
  US09/723,400
• Time in Patent Office: 1,653 Days
• Field of Search: 422/100, 422/99, 422/103, 422/102, 422/104, 422129-131, 738/633.3, 738/642.2, 141/236, 141/244, 436/180, 269/21, 134/21, 134/43, 134/44, 134/171
• US Class Current: 422/504
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01L 13/02 for receptacle or instruments
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 3/0244 using pins
• CPC Class Codes: B01L 9/52 Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 2035/00158 Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

The present invention is directed to systems and methods for producing microarrays of biological samples using an improved slide platform, vacuum manifold, and variable pin contact velocity. Slides onto which spotting pins produce the microarray of biological samples are secured on a flat slide platform using magnetic unit bars. The vacuum manifold effects optimal cleaning of the spotting pins by inducing increased air flow and turbulence. By reducing the velocity of the spotting pins before or after contact with the blot pad or the microarray slide, the morphology of biological samples deposited may be kept uniform. Similarly, low impact and separating velocities while acquiring biological materials prevent the pins from being overloaded with solution on the outside of the pin and causing sample wastage and non-uniform spotting.

Claims

1. The manifold assembly for removing liquid from in combination, a plurality of microarray spotting members, and a plurality of microarray spotting members, the spotting members each having a spotting member body and a first open end portion for printing a spot on a microarray slide, comprising:

a plate, the plate defining a plurality of fluid flow apertures extending through the plate, each aperture located to cooperate with a corresponding spotting member, each aperture having an axis a first diameter, an upstream edge forming an inlet and a downstream edge forming an outlet, the edges defining the aperture, the corresponding spotting member body having a second diameter wherein the second diameter is greater than the first diameter so that the spotting member may not entirely pass through the aperture, and wherein the first open end portion of the spotting member is adapted to extend into the corresponding aperture to a position where there is space between the spotting member and the aperture; and

turbulence means for creating turbulence in air flowing from the inlet to the outlet in the space between the spotting member and the aperture for removing liquid from the first open end portions of the spotting members.

2. The assembly of claim 1, wherein the turbulence means comprises i) aligning means for aligning the spotting members proximate to the apertures with each spotting member body extending generally axially away from the corresponding aperture, ii) reciprocating means for repeatedly reciprocating the spotting members generally axially toward and away from the apertures while maintaining the spotting members axially aligned with the corresponding apertures and limiting axially inward travel to provide clearance with the apertures in the limiting position and iii) vacuum means for drawing air around the spotting members to flow from the inlet to the outlet.

3. The assembly of claim 2, wherein the vacuum means comprises a source of vacuum and a structure for coupling the apertures to communicate with the source of vacuum to draw air around the spotting members to flow from the inlet to the outlet.

4. The assembly of claim 1, wherein the apertures comprise channels, the channels defined by the upstream and downstream edges.

5. The assembly of claim 4, wherein the apertures are arranged in parallel rows.

6. The assembly of claim 1 or 2, wherein the spotting members comprise pins.

7. The assembly of claim 6, wherein the pins are selected from the group consisting of solid pins and split pins.

8. The assembly of claim 1, comprising 48 apertures capable of simultaneously removing liquid from 48 spotting members.

9. The assembly of claim 1, comprising 32 apertures capable of simultaneously removing liquid from 32 spotting members.

10. The assembly of claim 4, further comprising a cover secured parallel to the plate over the inlets, the cover defining a plurality of cover apertures therethrough, each cover aperture concentric with an inlet of the plate, and the diameter of each cover aperture being less than the diameter of its concentric inlet.

11. The assembly of claim 1 or 2, wherein the first open end portion is tapered.

12. The assembly of claim 11, wherein approximately half of the tapered first open end portion is adapted to extend into the aperture.

13. The assembly of claim 2, wherein the first open end portion comprises a tip and the spotting member is reciprocable by the reciprocating means between first and limiting positions, the tip located outside the aperture in the first position and the tip located inside the aperture in the limiting position.

14. The assembly of claim 3, wherein the source of vacuum pressure provides a pressure of 50 to 90 psi.

15. The assembly of claim 3, wherein the source of vacuum pressure provides a pressure of 60 psi.

18. A method of removing liquid from a plurality of microarray spotting members, comprising creating turbulence in air flowing from the inlet to the outlet of the assembly of claim 1 or the microarrayer of claim 16 in the space between the spotting member and the aperture for removing liquid from the first open end portions of the spotting members wherein turbulence is created in air by:

applying a source of vacuum to the spotting members of the assembly to draw air around the spotting members to flow from the inlet to the outlet, aligning the spotting members proximate to the apertures with each spotting member body extending generally axially away from the corresponding aperture, repeatedly reciprocating the microarray spotting members generally axially toward and away from the apertures and limiting axially inward travel to provide clearance with the aperture in the limiting position to create turbulence in air flowing from the inlet to the outlet in the space between the spotting member an the aperture for removing liquid from the first open end portions of the spotting members.

19. The method of claim 18, wherein the spotting members are about 100 micrometers away from the inlet prior to reciprocation.

20. The method of claim 19, wherein the spotting members are reciprocated about a distance of 1 mm.

21. The method of claim 18, wherein each spotting member includes a tapered first open end portion adapted to extend into the aperture, the tapered first open end portion received in the aperture during at least part of the reciprocation.

22. The method of claim 21, wherein the first tapered open end portion reciprocates in and out of the aperture.

23. The method of claim 21, wherein the tapered first open end portion is spaced apart from the aperture during reciprocation.

16. A microarrayer, said microarrayer comprising:

a) a manifold assembly for removing liquid from a plurality of microarray spotting members;

b) a plurality of spotting members each having a spotting member body and a first open end portion for printing a spot on a microarray slide;

c) a plate, the plate defining a plurality of fluid flow apertures extending through the plate, each aperture located to cooperate with a corresponding spotting member, each aperture having an axis, a first diameter, an upstream edge forming an inlet and a downstream edge forming an outlet, the edges defining the aperture, a corresponding spotting member body having a second diameter wherein the second diameter is greater than the first diameter so that the spotting member may not entirely pass through the aperture, and wherein the first open end portion of the spotting member is adapted to extend into the corresponding aperture to a position where there is space between the spotting member and the aperture; and

d) turbulence means for creating turbulence in air flowing from the inlet to the outlet in the space between the spotting member and the aperture for removing liquid from the first open end portions of the spotting members.

17. The microarrayer of claim 16, wherein the turbulence means comprises i) aligning means for aligning the spotting members proximate to the apertures with each spotting member body extending generally axially away from the corresponding aperture, ii) reciprocating means for repeatedly reciprocating the spotting members generally axially toward and away from the apertures while maintaining the spotting members axially aligned with the corresponding apertures and limiting axially inward travel to provide clearance with the apertures in the limiting position and iii) vacuum means for drawing air around the spotting members to flow from the inlet to the outlet.

24. A method of removing liquid from a plurality of microarray spotting members, the spotting members each having a spotting member body and a first open end portion for printing a spot on a microarray slide, the liquid removed through a plate, the plate defining a plurality of fluid flow apertures extending through the plate, each aperture located to cooperate with a corresponding spotting member, each aperture having an axis, a first diameter, an upstream edge forming an inlet and a downstream edge forming an outlet, the edges defining a corresponding aperture, a corresponding spotting member body having a second diameter wherein the second diameter is greater than the first diameter so that the spotting member may not entirely pass through the aperture, and wherein the first open end portion of the spotting member is adapted to extend into the corresponding aperture to a position where there is space between the spotting member and the aperture, the method comprising:

first aligning the spotting members proximate to the apertures with each spotting member body extending generally axially away from the corresponding aperture;

drawing air around the spotting members to flow from the inlet to the outlet; and

repeatedly reciprocating the spotting members generally toward and away from the corresponding apertures while maintaining the spotting members axially aligned with corresponding apertures and limiting axially inward travel to provide clearance with the apertures in the limiting position,

wherein turbulence is created in air flowing from the inlet to the outlet in the space between the spotting member and the aperture wall for removing liquid from the first open ends of the spotting members through the apertures.

25. The method of claim 24, wherein the spotting member is reciprocated between first limiting positions;

the tip outside the aperture in the first position and the tip inside the aperture in the limiting position.