Cover member, method and treatment module for treating a biological sample on a substrate

Patent metadata

Field	Value
Publication number	US-10549271-B2
Application number	US-201214357888-A
Country	US
Kind code	B2
Filing date	Nov 15, 2012
Priority date	Nov 16, 2011
Publication date	Feb 4, 2020
Grant date	Feb 4, 2020

Abstract

A cover member for a substrate supporting a biological sample comprises first and second opposing ends, first and second opposing surfaces, a void in the second surface which, when juxtaposed with a substrate, forms a chamber, and a fluid inlet toward the first end and in fluid communication with the void. The void is bounded by void walls having one or more contoured regions for enhancing fluid movement within the chamber. A treatment module for a biological sample comprises the cover member, a support surface for a substrate bearing the biological sample and clamp means operable to releasably retain the cover member in juxtaposition with the substrate for an incubation period. A method for incubating the biological sample with one or more reagents uses the cover member.

First claim

The claims defining the invention are as follows:

1. A treatment module for a biological sample, the module comprising:
a cover member having;
i. opposing ends comprising a first end and a second end;
ii. opposing surfaces comprising a first surface and a second surface;
iii. a void in the second surface which, when juxtaposed with a substrate, forms a chamber; and
iv. a fluid inlet toward the first end and in fluid communication with the void;

a support surface configured to support the substrate and a biological sample thereon; and
a pivot arm configured to pivot the cover member about a dispersing edge causing fluid to move from the inlet along the substrate from the dispersing edge toward a fluid outlet, the pivot arm being operable to pivot the cover member to an open condition and to a closed condition, wherein
the pivot arm is operable to pivot the cover member and lift the cover member away from the support surface such that, of the first and second opposing ends, only one of the first and second opposing ends is lifted away from the support surface.

2. The treatment module according to claim 1 , comprising a robot configured to position one or both of the substrate and the cover member in the treatment module and optionally, wherein the robot is configured to dispense reagent into an inlet of the cover member during a treatment protocol.

3. The treatment module according to claim 1 , comprising a coupling operable to interchangeably connect one or more outlets, including the fluid outlet, of the cover member with one or more of a vent to atmosphere and a respective one or more negative pressure sources.

4. The treatment module according to claim 1 , further comprising a substrate retention member configured to retain the substrate on the support surface during opening of the chamber or separation of the cover member and the substrate.

5. The treatment module according to claim 1 , wherein the treatment module is configured for use with an automated sample processing instrument comprising a plurality of treatment modules operable independently under control of a controller of the instrument; and wherein operation of one or more of a clamp, a thermal exchanger, a robot, a negative pressure source and fluid dispenser is under the control of the instrument controller.

6. The treatment module according to claim 1 , further including a wash bay for exposing a surface of the cover member to a wash reagent, wherein optionally, the support surface is shaped to receive a substrate having a sample thereon and, in the absence of a substrate, to form the wash bay.

7. A method for incubating a biological sample with one or more reagents using a cover member according to claim 1 , including the steps of:
a. providing the sample on a substrate;
b. positioning the substrate and the cover member to form the chamber;
c. positioning a dispensing probe tip in mating contact with a fluid inlet of the cover member; and
d. driving a first volume of a first reagent into the inlet with force sufficient for the first reagent to substantially cover the sample on the substrate; and optionally
agitating reagent within the chamber;
e. agitating reagent within the chamber.

8. A method according to claim 7 , including one or more further steps comprising:
a. applying a negative pressure at a fluid outlet of the cover member to draw reagent within the chamber toward the outlet; and
b. removing the substrate from a support surface and immersing a second surface of the cover member in a wash reagent.

9. A method for incubating a biological sample with one or more reagents using a treatment module according to claim 1 , including the steps of:
a. providing the sample on a substrate;
b. positioning the substrate and a cover member to form a chamber;
c. positioning a dispensing probe tip in mating contact with a fluid inlet of the cover member; and
d. driving a first volume of a first reagent into the inlet with force sufficient for the first reagent to substantially cover the sample on the substrate; and optionally
e. agitating reagent within the chamber.

10. A method according to claim 7 , including one or more further steps comprising:
a. applying a negative pressure at an outlet of the cover member to draw reagent within the chamber toward the outlet and
b. removing the slide from a support surface of the treatment module and immersing a second surface of the cover member in a wash reagent.

11. The treatment module according to claim 1 wherein the cover member is removable from the treatment module.

12. The treatment module according to claim 1 wherein the cover member void is bounded by void walls having one or more contoured regions for enhancing fluid movement within the chamber.

13. The treatment module according to claim 1 , wherein the cover member includes:
the fluid outlet toward the second end and in fluid communication with the void; and
a guide at the inlet, configured to direct fluid into the inlet.

14. The treatment module according to claim 1 , wherein the cover member includes:
the fluid outlet, the fluid outlet being disposed toward the second end and in fluid communication with the void; and
the dispersing edge, the dispersing edge being disposed in fluid communication with the fluid inlet;
wherein the cover member is adapted to pivot about the dispersing edge causing movement of the fluid in the fluid inlet from the dispersing edge toward the fluid outlet.

15. The treatment module according to claim 1 , wherein the cover member includes a fluid dispersing feature from which fluid is dispensed, wherein the fluid dispersing feature is configured to dispense fluid from the inlet onto at least a width of the substrate.

16. The treatment module according to claim 1 , wherein the cover member further comprises the fluid outlet toward the second end and in fluid communication with the void and through which fluid may be withdrawn.

17. The treatment module according to claim 1 , wherein the cover member void is bounded by void walls having one or more contoured regions and the one or more contoured regions comprise rounded corners connecting side walls of the void with an end wall of the cover member to encourage fluid flow within the chamber.

18. The treatment module according to claim 1 , wherein the cover member void is bounded by void walls having one or more contoured regions and wherein the one or more contoured regions comprise a rounded taper connecting side walls of the void with an end wall of the cover member to encourage fluid flow within the chamber.

19. The treatment module according to claim 1 , wherein the cover member void is bounded by void walls having one or more contoured regions and wherein the one or more contoured regions comprise rounded cornices connecting the walls of the void with a void ceiling in the second surface of the cover member.

20. The treatment module according to claim 1 , wherein a void ceiling in the second surface of the cover member has a finish configured to enhance reagent propagation from the fluid inlet to the fluid outlet, and optionally, wherein the finish is at least one of etched, corrugated, dimpled, sloped, bowed, and rippled.

21. The treatment module according to claim 1 , wherein the cover member is disposable.

22. The treatment module according to claim 1 , wherein the cover member is formed from at least two parts including a cover member body and a cover member insert, wherein the cover member

Assignees

• Leica Biosystems Melbourne Pty

Inventors

• Dockrill Mark Brian
• Favaloro Anthony
• Ng Kenneth Heng-Chong
• Limon Martin
• Toogood Peter
• Bagnato Stephen John

Classifications

• B01L7/52

  with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples · CPC title

• B01L2200/0689

  Sealing · CPC title

• B01L2300/0877

  Flow chambers · CPC title

• Y10T436/25

  including sample preparation · CPC title

• B01L2300/1822

  using Peltier elements · CPC title