CryoCapsule microscopy specimen holder

CryoCapsule is a cyro correlative microscopy specimen holder that encapsulates the specimen between sapphire discs for live cell viewing, followed by high pressure freezing and electron microscopy.

High pressure freezing is the most advanced technology when it comes to vitrify a hydrated biological specimen while preserving the ultrastructure. Developed in the 80's [1], the technology evolved progressively to become accessible to a larger community. Still, the sample preparation prior to HPF remains tedious and often comes to advance expertise depending on the specimen [2].

The CryoCapsule is a new tool in the field of High Pressure Freezing (HPF) and correlative light and electron microscopy (CLEM). Comparable to a small petri dish, it is composed of a landmarked sapphire disc and a gold spacer ring (50um thick) maintained together by a plastic ring [3].

The specimens are encapsulated between the support sapphire disc (carbon landmarked) and a covering sapphire disc.

The CryoCapsule is loaded into a specific adapter (HPM010, HPM100, HPF compact 02). Live cell imaging is done directly on the specimen in the CryoCapsule prior to HPF.

CryoCapsule video

Post-HPF, the specimen is processed for freeze substitution [4] and room temperature sectioning.

CryoCapCell has developed a set of tools to manipulate the CryoCapsule [4] in most scientific environments.

References

1. Moor H, Bellin G, Sandri C, Akert K. The influence of high pressure freezing on mammalian nerve tissue. Cell Tissue Res [Internet] 1980 [cited 2013 Jul 24];209:201-16.
2. Mcdonald KL, Schwarz H, Thomas M, M?ller-Reichert T, Webb R, Buser C, Morphew M. "Tips and tricks" for high-pressure freezing of model systems. Methods Cell Biol [Internet] 2010 [cited 2011 Apr 19];96:671-93.
3. Heiligenstein X, Heiligenstein J, Delevoye C, Hurbain I, Bardin S, Paul-Gilloteaux P, Sengmanivong L, R?gnier G, Salamero J, Antony C, Raposo G. The CryoCapsule: Simplifying Correlative Light to Electron Microscopy. Traffic [Internet] 2014 [cited 2014 May 14];15:700-16.
4. Heiligenstein X, Hurbain I, Delevoye C, Salamero J, Antony C, Raposo G. Step by step manipulation of the CryoCapsule with HPM high pressure freezers. Methods Cell Biol [Internet] 2014 [cited 2014 Nov 27];124:259-74.

Decapsuleur and pencil - Tool to manipulate the CryoCapsule, correlative light and electron microscopy high pressure freezing cell, in most scientific environments.

Perforated petri dish - for CryoCapsule correlative light and electron microscopy high pressure freezing cell.