Bio System Installation

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Gregory Henry
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1 Bio System Installation Installation at the Danish Biobank, Copenhagen.

2 The Project In January 2012 Statebourne were awarded the Tender to equip the Danish National Biobank at the Statens Serums Institute. The Tender requirements were for the installation of fully automated liquid nitrogen refrigerators to store 2 million 1.4ml samples. The new refrigerators would be incorporated within an overall system which also included 7 existing older LN2 storage units and the combined system was to be supplied by a bulk liquid nitrogen tank sited externally and remotely (70 metres from the Cryo-room). Additionally a complete super-insulated vacuum pipeline and distribution management system capable of meeting a required LN2 consumption target of 25 litres per tank per day was required. A custom storage racking system for each refrigerator was also required for the safe and fast retrieval of 96 well storage plates. Finally, the system required high levels of sample and personnel safety monitoring to be provided to ensure that the required storage protocols were adhered to and that the cryo-room would provide a safe working environment for Staten Serums Institute personnel.

3 The Solution Providing 15 x Biosystem 100+ refrigerators which are off centre neck vessels that can store over 135,168 x 1.4ml samples in vapour phase with a low consumption of Ln2 A central line de-gas system that would degas the pipeline to the room quickly with all waste gas being piped away Individual hot gas by pass on every Biosystem, so that all the hot gas in the internal SIVL is removed right up to the back of the Biosystem resulting in less Ln2 gas in the room Daisy chain fill systems so the Biosystems fill on demand, Biosystems 100+ fill every 3-4 days so force filling will waste Ln2 Group filling, filling the Biosystems in groups to reduce the pressure on the ventilation system For the older tanks these were put on a separate filling system connected to a 500 litre slave tank. A fully integrated Oxygen alarm system that would link to ventilation system and have a connection to a slam shut valve to provide ultimate failsafe protection. RESULT Running 20 vessel (including 7 older units) over a 14 day period the consumption rate was 21 litres/vessel/day. Under the 25 litre/vessel/day target. All without an Oxygen alarm being activated

The Problem Cryoroom Supply Tank The external supply tank was positioned 70 metres from the Cryo-room, so there was 77 metres of super insulated vacuum line (SIVL) leading to the Cryo-room, and a

4 The Problem Cryoroom Supply Tank The external supply tank was positioned 70 metres from the Cryo-room, so there was 77 metres of super insulated vacuum line (SIVL) leading to the Cryo-room, and a further 40 metres of SIVL required in the room. It was estimated that just to cool the pipeline down about 70 Litres of Ln2 would be required and this would generate 42,000 litres of Warm N2 gas. Without this being dealt with the following issues would be: A very high consumption of Ln2 as any refrigerator would use at least 70 litres of Ln2 just to cool the pipeline, resulting in a consumption of perhaps 100 litres of Ln2/refrigerator/day. The additional hot gas coming into the room would deplete the Oxygen, resulting in the oxygen alarms always being activated and workers having to leave the room. The high amount of warm gas would infiltrate the refrigerators meaning that samples were Warmed on every fill resulting sample damage over time. The long fill times would also put the samples in danger since there is a higher chance that solenoids sticking open resulting in over-filling and possible sample -Ln2 contamination Again long fill times would result in moisture build up in room, with ice developing in refrigerators making sample extraction easy. Integrating seven older refrigerators, with older control systems into the cryo-room filling system. To integrate the ventilation system with the oxygen alarm so that it can cope with the hot gas generated during filling.

The Solution for the pipeline Insulated Hose Central Degas with phase seperator SIVL Waste Gas Line For the pipeline into the room the Biosystems were linked to a large degas valve and phase

5 The Solution for the pipeline Insulated Hose Central Degas with phase seperator SIVL Waste Gas Line For the pipeline into the room the Biosystems were linked to a large degas valve and phase separator this cleared all hot gas from the line quickly and cleaned the Ln2 so that there was just liquid entering the room. For the internal pipeline each Biosystem had a hot gas by pass fitted so that all the hot gas from the internal pipeline is removed right up to the back of the Biosystem. All waste gas was piped away. All connecting hose were insulated to reduce Ln2 consumption Biosystems were linked together to fill at the same time, and filled on demand this again reduced the consumption of Ln2 compared to a timer filing system Biosystems were filled in groups so that the Oxygen alarms and ventilation system were not swamped

Solution for Older Refrigerators 500 Litre tank with auto-fill Old Refrigerators SIVL de gas system For the seven older LN2 Refrigerators

system that worked on a timer so that there was always enough LN2 for the vessels to fill from.

system so that the minimum amount of hot gas went through these older refrigerators.

6 Solution for Older Refrigerators 500 Litre tank with auto-fill Old Refrigerators SIVL de gas system For the seven older LN2 Refrigerators it was decided that these could not be incorporated into a central filling system, so a 500 litre slave tank was fitted,with an auto-fill system that worked on a timer so that there was always enough LN2 for the vessels to fill from. The autofill system had a hot gas by-pass fitted to reduce consumption as much as possible, and the older tanks were linked into a de-gas system so that the minimum amount of hot gas went through these older refrigerators. Due to the age of the processors on these older refrigerators only batches of units could be linked filled. All this meant that the older tanks could work with the system, and use the minimum amount of LN2 but still be automatic.

7 Cryostor 500 with autofill system and hot gas bypass

Sample Storage The samples were stored in the Biosystem dry storage drum, so were completely free from LN2 contact and would be stored in temperatures of between -180 to -195 C, so the samples were

8 Sample Storage The samples were stored in the Biosystem dry storage drum, so were completely free from LN2 contact and would be stored in temperatures of between -180 to -195 C, so the samples were both thermally safe and protected from LN2 contamination. All the Biosystems are fitted with an overflow protection system that gives a further protection to the samples. For further peace of mind each Biosystem holds 270 Litre reservoir so samples will still be kept cold for over 15 days in the event of Ln2 supply failure. Sample Access Inside the storage drum a matrix was built to make rack location easy, and prevent any single racks falling over and being difficult to recover.

9 Access and Storage To make access easier for users movable steps were used. Each sample rack has a number plate with a colour to make identification easy.

The alternative is to start a fill process, and with this installation a fill activation can consume 100 litres of Ln2 on

10 De Fog system With the narrow necks of the archive units the Biosystems can be easily misted up. When a vessel is fitted with a hot gas bypass this will prevent an electronic de-fog system functioning. The alternative is to start a fill process, and with this installation a fill activation can consume 100 litres of Ln2 on every press of the button. So a simple additional de-fog system was fitted that the user could activate keeping the sample area clear of fog, but without wasting large volumes of Ln2.

11 Safety Central safety slam shut valve connected to an Oxygen alarm system Oxygen alarms are fitted throughout room. The Oxygen alarms will activate ventilation at any slight drop of Oxygen levels, but before the alarms are activated. When alarms are activated all users should leave room. If Oxygen levels drop further a central slam shut valve will isolate the Cryo-room from the main supply tank. The oxygen alarm will activate the central alarm system.

Data logging and alarm system Radio linked data logger Each Biosystem and older refrigerators are fitted with a local radio linked alarm system.

12 Data logging and alarm system Radio linked data logger Each Biosystem and older refrigerators are fitted with a local radio linked alarm system. This sends data back to a central data logging box that will record all storage temperatures and alarm conditions without the need for additional wiring. Records all storage temperatures. Records all alarm conditions for any refrigerators. Records all Oxygen alarms and conditions. If alarm happens users will be notified via a text message with time and alarm error. Central data logger can be interrogated from an external PC so the alarm can be checked and user can be assured on the type of alarm. Tel: +44 (0) Fax: +44 (0) cryobiology@statebourne.com

Bio System Installation

Bio System Installation Molecular Collection Facility, Natural History Museum London. The Project Installation of a complete Cryogenic storage facility for the storage of samples from endangered species.