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XtalTool – An all-in-one sample holder for crystallization, crystal soaking and data collection

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Protein crystals are often fragile and suffer from mechanical stress during manipulation and/or mounting. This can lead to impairment and even complete loss of diffraction quality. XtalTool was developed to minimize the handling steps of crystals in order to obtain the best possible diffraction data[1].


Clever and functional patented design[2-4]

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Feature Function Comments
A Round plastic support (outer frame) Support for airtight COC film (D) 22 mm or 18 mm diameter available
B Positioning aids Correct positioning of the XtalTool on the well of a crystallization plate
C X-ray transparent polyimide film with 5 µm pores Crystal support, allows diffusion of (soaking) solutions
  • Covers inner frame from inside
  • Oval in XtalTool HT and round in classic XtalTool
D Airtight transparent removable COC film Sealing of the well during crystallization and crystal manipulation
  • Covers entire XtalTool from outside
  • Pierced for crystal manipulation, resealing is possible
  • To be removed for data collection
E Extension Mimics pin and stabilizes XtalTool in the cryostream To be screwed into XtalTool Base for data collection
* Predetermined breaking points To break away the outer frame for data collection Only in XtalTool HT

How does it work?

1. XtalTool is used as a cover slide for hanging drop crystallization: The protein droplets are pipetted onto the yellow polyimide film, then XtalTool is flipped over and placed onto a greased well of a 24 well crystallization plate. Crystal growth is monitored using a standard light microscope.

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2. Crystal manipulation such as ligand soaking or cryoprotection is performed in a gentle way directly on the XtalTool. The crystal is never touched directly and the well remains closed, so any unnecessary disturbance of the crystal is avoided:

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A tiny hole is carefully pierced into the upper COC film using a fine needle.

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A fine paper wick is inserted into this tiny hole and remains in gentle contact with the polyimide film to suck away all the crystal surrounding liquid through the perforated polyimide film.

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New soaking solution is applied through this tiny hole in the gap, right on the opposite side of the crystal, it diffuses through the pores of the polyimide film and thereby reaches the crystal.

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The COC film is resealed by placing a protected finger on the hole and gently sliding across the puncture.


3. X-ray diffraction data are collected in-situ at ambient or cryo temperature: The COC film is pealed off and XtalTool HT is screwed into an XtalTool Base. The outer frame is then broken away at the predetermined breaking points and the crystal on the XtalTool HT can be flash frozen and stored in liquid nitrogen until data is collected.

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Watch the video and learn how to use the XtalTool sample holder to improve the X-ray diffraction data of protein crystals[1]:


Available products:

Product Cat. No. Amount/Content Description
XtalTool HT 22 mm X-XT-103 20 pcs.
  • Features predetermined breaking points at the outer frame
  • Compatible with robot assisted sample mounting
XtalTool HT 18 mm X-XT-104 20 pcs.
XtalTool Bases X-XT-105 20 pcs. B5-style goniometer bases customized for use with the XtalTool and XtalTool HT
XtalTool X-XT-101 24 pcs. + 1 Base
  • Has a fixed frame of 22 mm diameter
  • Manually mounted on the goniometer
XtalTool Soaking Kit X-XT-102 1 Soaking Stand
1 Needle Holder Pen
4 Test XtalTool Sample Holders
2 Sets of Paper Wicks
20 Cannulas
24 Long Extruded Pipette Tips
Contains all necessary equipment to perform a proper crystal soaking experiment on the XtalTool and XtalTool HT
Christin Reuter

E-Mail Christin for technical inquiries or further information: xtals@jenabioscience.com

References:

[1] Feiler et al. (2019) An All-in-one Sample Holder for Macromolecular X-ray Crystallography with Minimal Background Scattering. J. Vis. Exp. 149:e59722.
[2] Patent DE 10 2017 129 761.8, Christian G. Feiler, Dirk Wallacher, Manfred S. Weiss.
[3] Patent DE 10 2018 125 129.7, Christian G. Feiler, Dirk Wallacher, Manfred S. Weiss.
[4] Patent DE 10 2018 129 125.6, Christian G. Feiler, Dirk Wallacher, Manfred S. Weiss.