Sectioning volume samples

Patent metadata

Field	Value
Publication number	US-10054518-B2
Application number	US-201414533904-A
Country	US
Kind code	B2
Filing date	Nov 5, 2014
Priority date	Nov 5, 2013
Publication date	Aug 21, 2018
Grant date	Aug 21, 2018

Abstract

A microtome includes a blade located at an end of a trough that defines a cavity for holding a liquid; a sample block in which the at least one sample is suspended, the sample block is moveable relative to the blade such that when the sample block is passed across the blade a section is cut from the sample block; a plate that includes a support frame that defines an opening, and a transparent film extending across the opening, the transparent film being transparent to electrons, a grasper being configured to receive and retain the plate, wherein the grasper is moveable relative to the blade; and a pusher section that lacks the sample pusher section.

First claim

What is claimed is:

1. A method of cutting at least one sample suspended in a sample block of material, the method comprising:
passing the sample block across a blade one or more times;
with each passing of the sample block across the blade, cutting a sample section from the sample block, wherein one or more sample sections float on a liquid;
passing a blank block across the blade one or more times, wherein the blank block lacks the sample;
with each passing of the blank block across the blade, cutting a pusher section from the blank block, wherein one or more pusher sections float on the liquid and a last pusher section remains attached to the blade;
positioning a plate that includes an imaging region in the liquid and under at least the one or more sample sections;
clinging the one or more sample sections and the one or more pusher sections to the plate; and
removing the last pusher section from the blade.

2. The method of claim 1 , further comprising removing the plate from the liquid after the one or more sample sections and the one or more pusher sections are clung to the plate.

3. The method of claim 1 , wherein positioning the plate in the liquid and under the one or more sample sections comprises positioning the plate such that at least one of the samples of a sample section is positioned across the imaging region of the plate.

4. The method of claim 1 , wherein clinging the one or more sample sections and the one or more pusher sections to the plate comprises removing the liquid between the one or more sample sections and the one or more pusher sections and the plate.

5. The method of claim 1 , wherein clinging the one or more sample sections and the one or more pusher sections to the plate comprises lowering a level of the liquid relative to the plate so that all of the sections are lowered with the liquid onto the plate.

6. The method of claim 5 , wherein clinging the one or more sample sections to the plate comprises lowering the sample sections onto the plate such that the samples of the sample sections are positioned over the imaging region of the plate.

7. The method of claim 5 , wherein clinging the one or more sample sections and the one or more pusher sections to the plate comprises raising the level of the liquid before lowering the level of the liquid.

8. The method of claim 1 , wherein, with each passing of the sample block across the blade after the first pass of the sample block, after the sample section is cut from the sample block, the sample section sticks to the last sample section that was cut and is floating in the liquid.

9. The method of claim 1 , wherein the plate is positioned inside the liquid and under at least the one or more sample sections while the last pusher section remains attached to the blade.

10. The method of claim 9 , wherein removing the last pusher section from the blade comprises removing the last pusher section from the blade after at least one of the sample sections is clung to the plate.

11. The method of claim 1 , wherein the imaging region is a transparent region, the plate comprises a non-transparent region that provides a frame and defines the transparent region, and the transparent region includes a plastic film that extends across the transparent region and is secured to the non-transparent region.

12. The method of claim 11 , wherein clinging the one or more sample sections and the one or more pusher sections to the plate comprises clinging at least a portion of each sample section to the plate so that the sample in each sample section is adjacent the transparent region, and clinging at least a portion of the one or more pusher sections to the plate so that the pusher sections extend across the non-transparent region.

13. The method of claim 1 , wherein passing the sample block across the blade comprises passing the sample block across the blade one time such that one sample section floats on the liquid.

14. The method of claim 1 , wherein passing the blank block across the blade comprises passing the blank block across the blade one time such that one pusher section floats on the liquid.

15. The method of claim 1 , wherein passing the sample block across the blade comprises passing the sample block across the blade a plurality of times such that a plurality of sample sections float on the liquid.

16. The method of claim 1 , wherein passing the blank block across the blade comprises passing the blank block across the blade a plurality of times such that a plurality of pusher sections float on the liquid.

17. The method of claim 1 , wherein positioning the plate in the liquid and under the one or more floating sections comprises moving one or more sections and the plate with respect to each other such that the sections ride up a meniscus of the liquid that initially separates sections from the plate.

18. The method of claim 1 , wherein each sample section has a thickness of 20 nanometers to 2 micrometers.

19. A method of cutting a sample suspended in a sample block of material, the method comprising:
repeatedly passing a sample block across a blade located at the end of a trough that defines a cavity that holds a liquid;
with each passing of the sample block across the blade, cutting a sample section from the sample block, wherein a plurality of sample sections float onto the liquid held within the trough and remain adhered to each other;
passing a blank block across the blade one or more times, wherein the blank block lacks the sample, and with each passing of the blank block across the blade, cutting a pusher section from the blank block, at least one pusher section remaining attached to the blade;
clinging all of the plurality of sample sections to a plate that includes a support frame and an electron-transparent imaging region inside the support frame such that at least a sample in a sample section is positioned over the electron-transparent imaging region;
removing the pusher section that is attached to the blade from the blade;
detecting a relative physical feature between one or more of the blade, the trough, the liquid, the plate, and the sample block; and
controlling one or more of the passing, the cutting, the clinging, and the removing based on the detected relative physical feature.

20. The method of claim 19 , wherein detecting the relative physical feature between one or more of the blade, the trough, the liquid, the plate, and the sample block comprises one or more of:
detecting a distance between the blade and the sample block before the sample block is passed across the blade;
detecting an angle between the blade and the sample block before the sample block is passed across the blade;
detecting a distance between the plate and the blade; and
detecting an electrical feature between the plate and the liquid.

21. The method of claim 19 , wherein the pusher section is long enough to position the sample section over the electron-transparent imaging region of the plate.

22. The method of claim 19 , further comprising positioning the plate that includes the imaging region in the liquid and under the one or more floating sample sections while the pusher section remains attached to the blade.

23. The method of claim 19 , wherein removing the pusher section from the blade comprises removing the pusher section from the blade after at least one of the sample sections is clung to the plate.

24. The method of claim 19 , wherein clinging the sample sections to the

Assignees

• Hughes Howard Med Inst

Inventors

• Hess Harald F
• Peale David
• Lee Patrick R

Classifications

• Y10T83/6577

  With means to adjust additional means · CPC title

• G01N1/06 Primary

  providing a thin slice, e.g. microtome · CPC title

• G01N2001/065

  Drive details · CPC title

• Y10T83/148

  Including means to correct the sensed operation · CPC title

• Y10T83/0448

  With subsequent handling [i.e., of product] · CPC title