Xenon Chamber User Guide
Contents
1. Xenon Chamber HAMPTON RESEARCH Solutions for Crystal Growth Table of Contents Introduction 2 Xenon 2 What is Included 2 Available Separately 2 Before Getting Started 3 Getting Familiar 4 Installation 5 Operation Requirements 6 Pre Operation 6 Operation 8 Shutting Down the Xenon Chamber 10 General Information 11 Specifications 1l Recommended Accessories 11 Important Safety Instructions 1 Warning Il References 12 Service and Support 12 Tips and Troubleshooting 13 Alignment of the Plunger 13 Alignment of the Chamber Lock 13 Warranty Policy 14 Product Reference Information 14 section The Xenon Chamber Introduction The Xenon Chamber is a pressure chamber designed to produce xenon derivatives of biologi cal macromolecular crystals Typically isomorphous derivatives are formed by diffusing heavy atoms such as lead or mer cury based compounds into the crystal with the anticipation that the heavy atom molecules will bind in an ordered fashion to each biological macromolecule Xenon Xenon is a noble gas which can bind to specific sites in a biological macromolecule There are numerous examples in the literature demonstrating that xenon macromolecule complexes can serve as heavy atom derivatives see page 11 The xenon macromolecule complex is obtained by placing the macromolecular crystal under a pressurized xenon atmosphere The use of xenon in a pressurized chamber allows one to manipulate th2. over the Dewar Step 9 Lower the Plunger and align the Dewar so that the CrystalCap Magnetic or CrystalCap Magnetic Copper here after referred to as CrystalCap aligns flush with the Magnetic Base on the bottom of the Plunger Step 10 Raise the Plunger and place the CrystalCap onto the Magnetic Base of the Plunger Step 11 Slide the Plunger with the CrystalCap to the left against the Left Guide Stop Screw over the Xenon Chamber Step 12 Lower Plunger into the Xenon Chamber Step 13 Close the Chamber Lock completely The chamber is now sealed and ready to accept xenon Step 14 Close Safety Shield Step 15 Check to see that all valves in the xenon tank and Xenon Pressure Regulator are closed Step 16 Turn the 3 Point Valve to the Apply Pressure position Step 17 Open valve on xenon gas cylinder Step 18 Open Pressure Regulator Valve by turning the T Handle clockwise to a pressure between 0 and 600 psi Step 19 Slowly turn the Black Needle Valve counter clockwise to set pressure in the Xenon Chamber This pressure can be monitored by the Chamber Pressure Gauge on the Chamber Platform Do not allow pressure to go beyond 600 psi Note If the pressure of the Xenon Chamber exceeds 600 psi pressure will vent through the Safety Valve Step 20 Once the desired pressure has been reached clo
3. 24 931 935 Service and Support For service and support of the Grease Machine please contact Hampton Research Please read and understand the Warranty Policy for your Xenon Chamber prior to contacting Hampton Research 12 p section Appendix B Tips and Troubleshooting Alignment of the Plunger Plunger alignment can be adjusted by modifying the either the Left Guide Stop Screw or Right Guide Stop Screw If the Plunger does not align properly over the dewar even after trying to reposition the dewar under the Plunger adjust the Right Guide Stop Screw by loosening the nut Adjust the Right Guide Stop Screw as appropriate and tighten the nut If the Plunger does not align properly over the Chamber Lock adjust the Left Guide Stop Screw by loosening the nut Adjust the Left Guide Stop Screw as appropriate and tighten the nut Alignment of the Chamber Lock If a problem persists where the fins of the Plunger do not align properly with the fins of the Chamber Lock adjust the Chamber Lock Stop Screw by loosening the nut Adjust the Chamber Lock Stop Screw as appropriate and tighten the nut 13 Warranty Policy Please complete and return the enclosed warranty validation paper This is your proof of purchase and warranty activation Failure to return the enclosed warranty validation paper will forego any and all rights to warranty prote
4. Legend 10 Left Guide Stop Screw 1 Dewar Dock 11 Guide Ko 2 Chamber P latform e ae i 3 Chamber Pressure Gauge i dad andie 4 Slow Release Gas Valve Plunger 15 Magnetic Base 5 3 Way Valve 16 Right Guide Stop Screw 6 Xenon Chamber 7 Chamber Lock 17 Safety Shield 18 Safety Lock 8 Lock Lever Stop Screw 9 Lock Lever 19 Inlet y section Quick Connector Attaches to Xenon Chamber Inlet Operating the Xenon Chamber Installation Tools Required Crescent Wrench Prior to installation of the Xenon Pressure Regulator check to see that the xenon tank valve is closed be sure the Pressure Regulator Valve is closed by turning the T Handle counter clockwise and that the Black Needle Valve is closed by turning it clockwise Installation and setup of the Xenon Chamber should be done on a sturdy flat surface within 20 inches of the xenon tank Regulated Pressure Gauge Xenon Tank Gauge Pressure Regulator Valve T Handle Black Needle Valve Attach to Xenon Gas Tank Xenon Chamber Pressure Regulator Step 1 Attach Xenon Chamber Pressure Regulator Catalog Number HR4 793 to the xenon tank Tighten using a crescent wrench Step 2 Attach other end of the Xenon Chamber Pressure Regulator to the Xenon Chamber Inlet Pull back on the quick connector to expose the metal ball bearings While pulling back on the quick connector attach to the male inlet on the Xenon Chamb
5. T Handle clockwise Step 7 Slowly open the Black Needle Valve by turning it counter clockwise This will release any remaining pressure within the Xenon Chamber Pressure Regulator through the Xenon Chamber Step 8 Disconnect the Xenon Chamber Pressure Regulator from the Xenon Chamber Inlet Pull back on the quick connector to expose the metal ball bearings This will cause an auto matic release of the quick connector from the male inlet on the Xenon Chamber Step 9 Disconnect Xenon Chamber Pressure Regulator from the xenon tank 10 y section Appendix A General Information Specifications Operation Pressure 0 600 psi 0 41 0 41 bar Pressure Regulator CGA Number 580 Pressure Regulator Hose Length 21 inches Recommended Accessories CrystalCap Magnetic CrystalCap Copper Magnetic 1000 ml Dewar Catalog Number HR4 699 1000 ml Dewar Cork Cap Catalog Number HR4 701 Replacement Mini Vial with Wick Catalog Number HR4 795 Xenon Chamber Pressure Regulator Catalog Number HR4 793 Xenon Recovery System Catalog Number HR4 797 Important Safety Instructions For your own safety and that of your equipment read and follow all the instructions in this section Keep these instructions available for reference by you and others Warning D jar D L jar se of this equipment requires working with pressurized xenon gas Only personnel famil with
6. 00m Dewar The CrystalCap holding the crystal will be lowered into the CrystalCap Vial containing liquid nitrogen or liquid propane Step 19 Lift the Magnetic Release Lever This will release the CrystalCap from the Magnetic Base on the bottom of the Plunger Raise the Plunger while lifting the Magnetic Release Lever and slide the Plunger away The crystal is now derivatized hopefully and frozen and is now ready for data collection Shutting Down the Xenon Chamber Once xenon derivatization has been completed depressurization is required in order to discon nect the xenon tank from the Xenon Chamber Step 1 Close the Tank Valve on the xenon tank Close the Pressure Regulator Valve by turning the T Handle counter clockwise until it spins freely Close the Black Needle Valve clockwise until it stops Step 2 Set the 3 Point Valve to Release Pressure Turn the valve to release any pressure in the Xenon Chamber Step 3 Open Chamber Lock raise Plunger and slide it to the right away from the Xenon Chamber Step 4 Remove the Mini Vial with Wick from the Xenon Chamber Step 5 Turn the 3 Point Valve to Apply Pressure with the Xenon Chamber open This will allow any pressure in the hose connecting the xenon tank to the Xenon Chamber to be released Step 6 Slowly open the Pressure Regulator Valve by turning the
7. ction and you will be subjected to standard repair costs of the product in question If you do not agree with the following terms and con ditions please contact Hampton Research Customer Service for return information Hampton Research Corp hereafter HR warrants for ninety 90 days from date of purchase by the initial purchaser that this product shall be free of defects in material and workman ship Any material or workmanship determined defective by HR within the ninety 90 days shall be repaired or replaced without charge for parts and labor provided the unit is returned in the original box and transportation costs prepaid to Hampton Research 34 Journey Aliso Viejo CA 92656 3317 or to such other service facility authorized by HR HR shall pay shipping costs to return the unit to its owner Products returned for repair must first be approved for return by Hampton Research Contact Hampton Research for a Repair Return Authorization Number Products returned without a Repair Return Authorization Number will not be accepted This warranty does not cover damage caused by accident misuse abuse neglect unauthorized or improperly performed repairs or alterations and or wear and tear occasioned by the use of the product and does not include any expense for inconvenience or loss of use while the product is being repaired or replaced HR expressly disclaims any liability for consequential damages arising from the sale use or inability to
8. e occupancy and binding of xenon by varying pressure and incubation time What is Included Included in this package are the following Quantity 1 Xenon Chamber Quantity 1 CrystalCap Holder for 1000ml Dewar Quantity 5 Mini Vial with Wick Available Separately Xenon Chamber Pressure Regulator amp Connector Catalog Number HR4 793 Xenon Recovery System Catalog Number HR4 797 Mini Vial With Wick 5 Pack Catalog Number HR 795 Before Getting Started Operators of the Xenon Chamber should be trained in the use and safety of a pressurized gas cylinder xenon gas tank Please use ANSI OSHA approved safety goggles and any other safety equipment recommended by Local State and Federal government regulations Hampton Research shall not be held liable for any damage resulting from handling or from contact from said product Please read all instructions carefully This apparatus is only to be used with inert gases It is recommended that the Xenon Chamber Pressure Regulator amp Connector Catalog Number HR4 793 be used with the Xenon Chamber Exact fittings cannot be guaranteed with other pressure regulators Please refer to operation specifications in Appendix A of the User Manual Getting Familiar The illustrations below provides an overview of your Xenon Chamber Please familiarize yourself with the Xenon Chamber prior to beginning the installation procedure Front View Side View
9. er Attachment will be confirmed by the quick connector clicking firmly onto the male inlet Operation Requirements The following items are necessary for the use of the Xenon Chamber 1 Xenon Gas 2 Liquid Nitrogen CrystalCap Magnetic CrystalCap Copper Magnetic 4 MicroTube amp CryoLoop or Mounted CryoLoop 5 1000ml Dewar 6 Tweezers Pre Operation Pre operation is recommended in order to determine the proper setting conditions and for aligning the dewar on the Dewar Dock This should be done simply using a CryoLoop with only mother liquor suspended within the CryoLoop no crystal Please use ANSI OSHA approved safety goggles and any other safety equipment recommended by Local State and Federal government regulations Step 1 Obtain a 1000ml Stainless Steel Dewar Flask Step 2 Place the CrystalCap Holder inside the Dewar Step 3 Place a CrystalCap Vial into the CrystalCap Holder Step 4 Set Dewar onto the Dewar Dock Step 5 Place approximately 100 microliters of mother liquor into a Mini Vial with Wick Shake the Mini Vial to allow the mother liquor to absorb into the wick Add additional mother liquor as necessary to maintain approximately 2 millimeters of mother liquid in the Mini Vial Step 6 Open Safety Cover Step 7 Place the Mini Vial with Wick into the Xenon Chamber Step 8 Raise the Plunger and slide it to the right against the Right Guide Stop Screw
10. hen close the Safety Shield The chamber is now sealed and ready to accept xenon Step 9 Turn the 3 Point Valve to the Apply Pressure position Step 10 Open valve on xenon gas tank to obtain a desired pressure then close valve on xenon gas tank Step 11 Open Pressure Regulator Valve to a pressure between 0 and 600 psi Step 12 Slowly turn the Black Needle Valve to set the desired pressure in the Xenon Chamber This pressure can be monitored by the Chamber Pressure Regulator on the Chamber Platform Do not allow pressure to go beyond 600 psi Step 13 Once the desired pressure has been reached close the Black Needle Valve and turn the 3 Point Valve to Hold Pressure Monitor the Chamber Pressure Gauge during incuba tion to assure pressure is maintained at the desired level Incubation time will vary depend ing upon the protein Typical incubation time is 15 minutes to 60 minutes Step 14 After incubation slowly turn the 3 Point Needle Valve to the Release Pressure posi tion Pressure will be released at the desired rate as determined in the pre operation setting of the Slow Release Gas Valve Step 15 Once pressure has been released open the Safety Shield Step 16 Open the Chamber Lock Step 17 Raise the Plunger and slide it to the right over the dewar and CrystalCap Vial con taining liquid nitrogen Step 18 Lower Plunger into the 10
11. n Chamber is already primed Once STEP 16 is reached again turn the 3 Point Valve SLOWLY Pressure will be applied to the Xenon Chamber immediately Please use caution Operation Begin actual operation of the Xenon Chamber only after going through the Pre Operation set up Note When beginning the Operation procedure after the Pre Operation procedure the hose connecting the xenon tank to the Xenon Chamber is already primed Once STEP 8 is reached turn the 3 Point Valve SLOWLY Pressure will be applied to the Xenon Chamber immedi ately Please use caution Step 1 Fill the 1000ml Dewar with liquid nitrogen Be sure that liquid nitrogen has also been placed within the CrystalCap Vial Step 2 Place approximately 100 microliters of mother liquor into a Mini Vial with Wick Shake the Mini Vial to allow the mother liquor to absorb into the wick Add additional mother liquor as necessary to maintain approximately 2 millimeters of liquid in the Vial Step 3 Open the Safety Shield Step 4 Place the Mini Vial with Wick into the bottom of the Xenon Chamber Step 5 Raise the Plunger and attach CrystalCap with crystal to the Magnetic Base of the Plunger Step 6 Slide the Plunger with the CrystalCap to the left against the Left Guide Stop Screw over the Xenon Chamber Step 7 Lower Plunger into the Xenon Chamber Step 8 Close the Chamber Lock t
12. se turn clockwise the Black Needle Valve and turn the 3 Point Valve to Hold Pressure Step 21 After one minute turn the 3 Point Valve to the Release Pressure position It is rec ommended that pressure be released over a 3 to 5 second period The at which pressure is released can be controlled by using the Slow Release Gas Valve Note Pressure is ONLY released from the Xenon Chamber not from the hose connecting the xenon tank to the Xenon Chamber Step 22 Once pressure has been released open the Safety Shield Step 23 Open the Chamber Lock Step 24 Raise the Plunger and slide it to the right Step 25 Lower Plunger into the CrystalCap Vial Raise the Magnetic Release Lever and raise the Plunger to leave the CrystalCap in the CrystalCap Vial At this point confirm the align ment of the Dewar and the CrystalCap holder with the Plunger Step 26 Remove the CrystalCap and observe the CryoLoop under a microscope The proper operation of the Xenon Chamber that is the speed at which pressure was increased and decreased should leave the mother liquor within the CryoLoop Rapid pressur ization and depressurization of the Xenon Chamber will cause the mother liquor to blow off the CryoLoop Please practice several times prior to the actual use of a crystal Note When repeating the Pre Operation procedure the hose connecting the xenon tank to the Xeno
13. the use of pressurized xenon gas should handle this device se ANSI OSHA approved safety goggles and any other safety equipment recommended by cal State and Federal government regulations se of the Xenon Chamber requires use of liquid nitrogen Personnel should also be famil with the use and handling of liquid nitrogen 11 References Machius M Henry L Palnitkar M Deisenhofer J A Xenon Derivative of UvrB Crystal Prepared Using the Hampton Research Xenon Chamber 1999 Proc Natl Acad Sci USA 96 11717 11722 Otwinowski Z amp Minor W 1997 Methods Enzymol Vol 276 edited by C W Carter Jr and R M Sweet New York Academic Press Sauer O Schmidt A Kratky C Freeze trapping isomorphous xenon derivatives of protein crystals J Appl Cryst 30 476 Schiltz M Fourme R Broutin amp Prange T 1995 Structure 3 309 316 Schitz M Prange T amp Fourme R 1994 J Appl Cryst 27 950 960 Schiltz M Shepard W Fourme R Prange T De La Fortelle R amp Bricogne G 1997 Acta Cryst D53 78 92 Soltis S M Stowell Wiener MC Phillips Jr amp Rees D C 1997 J Appl Cryst 30 190 194 Stowell Soltis SM Kisker C Peters J W Schindelin H Rees D C Cascio D Beamer L Hart PJ Wiener M C amp Whitby 1996 J Appl Cryst 29 608 613 Vitali J Robbins AH Almo C amp Tilton RF 1991 J Appl Cryst
14. use the product Any warranty implied by law including any warranty of mer chantability or fitness is expressly limited to the ninety 90 days warranty term The fore going statements of warranty are exclusive and in lieu of all other remedies Product Reference Information Purchase Date Invoice Number Serial Number Please save the following information and have it ready at the time of any questions or con cerns regarding the Xenon Chamber 2001 2003 Hampton Research Corp all rights reserved Printed in the United States of America This guide or parts thereof may not be reproduced in any form without the written permission of the publishers 14
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