Project Title. Department Address Medma-Account Telephone

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1 Project Title User First- and Surname Department Address Medma-Account Telephone I have read the terms and conditions of the Core Facility LIMA and will comply with them Date User Signature Department Head First- and Surname Date Signature Head of Department Cost Unit Alternative Cost Unit User Policy Core Facility LIMA

2 User Policy Live Cell Imaging Mannheim, LIMA at CBTM Version 2.0 March 2013 Responsible persons: Dr. Wolfgang Greffrath, C4 411, Tel. ( ) 9933; Scientific support and Management of the Core Facility LIMA Dr. Julia Bucher, C , Tel. ( ) 9937; Technical support of instruments, Laser safety Officer Handan Mörz, C2 206, Tel. ( ) 9938; Technical support sample preparations Sigrid Hugues, Dipl-BW (FH), C4 404, Tel. ( ) 9928 Administrative support 1. PREAMBLE: The Core Facility Live Cell Imaging Mannheim, LIMA offers confocal and multiphoton laser scanning microscopy related services at the Center of Biomedicine and Medical Technology Mannheim (CBTM) of the Medical Faculty Mannheim. LIMA was founded through funding by the Structure- and Innovation Fond for research, Baden- Württemberg (SI-BW) given to the Chair Prof. Dr. Rolf-Detlef Treede, as well as by two applications for major research devices according to article 91 b GG of the DFG ( principal applicants Treede/Greffrath; side applicants Professors Augustin, Boutros, Klüter, Schubert, Schultz and Sleeman). The Core Facility LIMA is located on the ground floor of the Tridomus building,house C, Ludolf-Krehl-Street 13-17, Mannheim, in rooms C1 103 and 104. At the time of foundation, LIMA s equipment includes an inverse laser scanning microscope (LSM) Leica SP5 Mid multi-photon system (UV-Vis-IR), an upright Leica LSM SP5 DS as well as a cell culture hood and further instruments; a detailed list of devices can be found in the attachment 1. LIMA seeks for constant improvement, modification and expansion of the existing laser scanning microscopes towards the current state of the art. 2. MANAGEMENT The Core Facility LIMA is headed by Dr. Wolfgang Greffrath and Dr. Julia Bucher of the Chair of Neurophysiology at the CBTM of the Medical Faculty Mannheim of User Policy Core Facility LIMA Page 1 of 7

3 Heidelberg University. Prof. Dr. Rolf-Detlef Treede is point of contact for users from the industrial field. The advisory board consults LIMA s management and represents the interests of the users, to whom the following members belong: 1. Applicants of research instrument proposal (see Preamble) as long as they are members of Heidelberg University 2. Executive Director of the CBTM 3. Medical Faculty Mannheim in the person of the Faculty s Managing Director The advisory board holds meetings together with LIMA s management on a regular basis in order to report on incidents and/or changes within the administrative structure. Should a member of the advisory board retire from the Board, the remaining members decide on who will possibly be appointed to the Advisory Board. 3. PREFACE 3.1 Services The Core Facility LIMA offers the following services: a) Training in the operation of the microscopes b) Consultation on experiments of the confocal laser microscopy, multi-photon microscopy and living cell microscopy c) Management of microscope time and invoicing e) Development and establishment of microscopic methods in cooperation with the users f) Maintenance of microscopes and computers g) Planning, organization and implementation of experiments at the microscopes against payment 3.2 User Fee LIMA charges user fees from the users for the above-namedservices in order to cover running costs (e.g. consumables, such aslens cleaning tissue, immersion media; instrument add-ons like new filters; evaluation software and its updates, etc.). Scope and amount are calculated on a personal and project basis as well as on the type of services used. Charges for usage and additional services are determined and adapted by the management of the Core Facility LIMA. Attachment 2 shows the actual breakdown and listing of user fees. 3.3 Annual report LIMA s management issues an annual report, keeping the advisory board informed about the most important events, activities and developments of the Core Facility. User Policy Core Facility LIMA Page 2 of 7

4 4. USAGE 4.1 Authorized users On informal request, user permission is given by LIMA to: 1. Employees from work groups of the co-applicants of the research instrument proposal 2. Members of the Medical Faculty Mannheim of University of Heidelberg and other scientific institutions 3. If applicable, external users from the industrial field Prerequisite for the user permission is a successfully completed training session. Permission shall be granted individually by LIMA s management after completion of training; authorized users are enabled to login at the instruments` computer and the booking-calendar labtima by activation of their medma-account. User registration and activation of the medma-account are prerequisites for working as an authorized user at LIMA`s instruments; only authorized users have access to rooms and instruments. It is explicitly forbidden to login at the instruments computer for persons, who have not been trained or other non-authorized persons, and may lead to the immediate cancellation of the user permission. The user authorization is valid for one year and may be extended on informal written request. 4.2 Hours of operation and usage Generally, LIMA is available to authorized usersaffiliated to the CBTM, 24 hours per day and 7 days per week. Access for all other users is limited to a core time on weekdays between 7 a.m. and 7 p.m. Support is provided by responsible persons at LIMA on weekdays at the usual working time. Microscope times are assigned by the following priority: 1. Members of work groups of applicants 2. Employees of the Medical Faculty Mannheim 3. Employees of the University of Heidelberg 4. Other groups 4.3 Booking and Invoicing Booking of the Core Facility s instruments is made electronically via labtima In addition to the electronic booking, each user needs to record microscope times and further information (lamp burning hours, lasers used, genetically modified organism, etc.) in an instrument-specific lab book. The maximum booking time is 4 hours during the day between 7a.m. and 7p.m.; longer booking hours are possible outside of this time, e.g. for time lapse series. User Policy Core Facility LIMA Page 3 of 7

5 Bookings made longer than 4 weeks in advance require prior permission of the management; if needed, additional individual exceptions can be made. Each user shall not make more than three bookings per week. Serial consecutive bookings done by various users belonging to the same research group shall be avoided. Generally, double bookings of both LIMA instruments are not allowed. Deviations from this procedure may be agreed on in individual cases. Electronic booking is a pre-requirement for the use of instruments and serves as a basis for the calculation of user fees. In addition, electronic bookings are synchronized by the LIMA`s management with the lab book notes at the microscopes. Free cancellation is possible at any time up to 24 hours before the booked microscope time. Subsequently, the booked period will be charged, insofar as no replacement can be provided by the user. If another user wants to take over your booking, the required rebooking can be done by a responsible person. According to agreement and availability, an additional specific objective or experiment-specific equipment may be booked in labtima. The exchange of objectives is exclusively made by a person in charge at LIMA. Extensive modifications at the instruments have to be announced and paid separately as remodeling time and can only be done together with a person in charge at LIMA. 4.4 Rights and duties The users are obliged to know LIMAs rules, comply with them and follow the instructions of persons in charge. With their signature they confirm knowledge of these rules at the time of their training session (see cover page). User rights: Access to microscope times at LIMA s instruments Training for the operation of the microscopes Scientific advice regarding experiments Development and establishment of microscopic methods in cooperation with the management Laser safety instruction once a year Annual report (see section 3.3) User duties: Each user is obligated to follow the practical instructions listed in attachment 3 as part of the terms & conditions to ensure proper operation. Each user is responsible for the proper use of the instruments during the booked time and is responsible for any damages caused. Damages or functional limitations of the instruments, which already exist at the beginning of User Policy Core Facility LIMA Page 4 of 7

6 the booked time, shall be reported immediately, later complaints cannot be accepted. Upon booking the user is obligated to pay the user fee. An exception is made for bookings which are cancelled at least 24 hours prior to the use in case of the microscope s malfunction, provided that the malfunction is not caused by the user. Correct registration does not guarantee any legal rights for microscope time. In case of actual or impending functional restrictions of the instruments, unforeseen or urgent repair work or maintenance etc., the instruments may have to be restricted or shut down at short notice the decision shall be made solely by a responsible person. The user is obligated to make a note in the enclosed lab book and to sign his notes. In particular, all errors of the instruments must be documented. Furthermore, error must be reported immediately (via and/or by phone). The user has to leave the workplace tidy and clean. The user is responsible for the correct attention of the corresponding guidelines of the general and specific laboratory rules, the ordinance on Biological Agents, the Gene Technology Law and other work and safety-related rules and regulations, compliance with these rules and regulations is presumed (see attachment 4). The user is not allowed to change microscope components and/or install any software. Removal of instruments, objectives and other microscope equipment for use outside of LIMA is strictly forbidden. The user is obligated to appreciate LIMA by citation in the acknowledgements of scientific publications, as well as in Bachelor s -, Master s and, Doctoral theses as soon as pictures, data and analyses are shown, which were generated in LIMA. The respective DFG s project number must be listed as well. For the upright microscope: We acknowledge the support of the Core Facility Live Cell Imaging Mannheim at the CBTM (DFG INST 91027/9-1 FUGG) For the inverse Multi-Photon microscope: We acknowledge the support of the Core Facility Live Cell Imaging Mannheim at the CBTM (DFG INST 91027/10-1 FUGG) Once a publication carries the status in press, a PDF version of the publication must be sent to: julia.bucher@medma.uni-heidelberg.de Payment of the user fee does not exclude co-authorship in a publication. If one of LIMA s persons in charge contributes a substantial intellectual and/or experimental contribution to the publication, he/she has to be treated like any other co-author. User Policy Core Facility LIMA Page 5 of 7

7 It is in LIMA s interest that all users generate and publish pictures and data at the instruments with the highest quality standards. Users are obligated to follow the management s instructions and suggested improvements to this effect. LIMA s management is entitled to ask for compliance of good scientific practice and, to this end, provide users with the mandatory guidelines. In this context the following publication shall be mentioned: Cromey DW Avoiding Twisted Pixels: Ethical Guidelines for the Appropriate Use and Manipulation of Scientific Digital Images of the journal Science and Engineering Ethics 2010 Dec;16(4):639-67; Springer publishing company. 5. DATA STORAGE Data may be temporarily stored locally on the computer of the microscope. Data can be saved by the user on drive (D:) in a specific file, which allows precise assignment. The user is obligated to transfer his data as soon as possible from the hard disk drive to his own medium and to delete his data from the hard disk drive. In case of acute shortage on the local computer, data older than one week will be deleted without further investigation. Until further notice, regular backups of the microscopic computer data will not be performed. LIMAs persons in charge take no responsibility for the user s data. 6. INVOICING, USE OF USER FEES User fees charged will be posted on the Core Facility LIMA s own account ( ). Charging is made quarterly in accordance with the actual user fee listing (see attachment 2). Current expenses of less than 1000 for consumables and microscope components can be paid by LIMA s persons in charge through its account. Expenses of more than 1000 must be paid in equal parts by the work groups who require those components. In any case, the Core Facility LIMA will be added as one user. The expense for each interested user is calculated as follows: X*1/(n+1); X = Amount, n = number of users +1 LIMA After purchasing, the component becomes LIMA s property and remains within the Core Facility. All extensions at the instruments must be ordered and fixed by a responsible person. Repairs and maintenance will be performed, wherever possible, by persons in charge. Extensive repairs require a manufacturer`s technician, whose commission is carried out only by a responsible person. A service contract with the manufacturer exists for the instruments. User Policy Core Facility LIMA Page 6 of 7

8 Management of the Core Facility LIMA in March 2013 Date Chair of Neurophysiology Date Core Facility LIMA Date Executive Director CBTM Date Medical Faculty Mannheim User Policy Core Facility LIMA Page 7 of 7

9 Attachment 1: Technical Equipment Inverse Leica SP5 Mid Multi photonsystem (UV-Vis-IR) - Microscope DMI 6000CS Bino equipped with bright field, phase contrast and interference contrast - External fluorescent lamp EL Filter system for epifluorescence: A (BP nm, LP 425 nm); I3 (BP nm, LP 515nm); N2.1 (BP nm, 590 nm); TX2 (BP 560/40 nm, LP 645/75 nm) - Available laser lines: Ar-Laser (568 nm/5 mw; 476 nm/5 mw; 488 nm/20 mw; 496 nm/5 mw; 514 nm/20 mw), DPSS (561 nm/20 mw), HeNe (594 nm/2,5 mw, 633 nm/10 mw) - Infrared Ti: Saphire Laser MaiTai Deep Sea (variable nm/2,5 W) - 8 channel AOTF; AOBS (0,6-2 nm selectivity) - Scanning table, Super-Z Galvo stage - Variable scanner: frame scan: up to 25 frames/s; line scan: up to 2800 lines/s - Resonance scanner: frame scan: up to 250 frames/s; Line scan: up to lines/s - 1 TLD-Detector for recording of transmission light images - 4 Internal detector channels for recording of fluorescent images - 2 External detector channels for MP - Objectives: 5x/0.15 NA air; 10x/0.30 NA air; 20x/0.7 NA air; 25x/0.95 NA water; 40x/1,3 NA oil; 40x/1,1 NA water - Tempered Ludin incubation chamber with gas mixer (gas concentration selectable 1-15% CO 2 ); Aerated mini-incubator for Z-Galvo table Upright Leica LSM SP5 DS - Microscope DM 6000 B - External fluorescent lamp EL Filter system for epifluorescence: I3 (BP nm, LP 515 nm); N2.1 (BP nm, 590 nm) - Available laser lines: Diode laser (405 nm/50 mw), Ar-laser (458 nm/5 mw; 475 nm/5 mw; 488 nm/20 mw; 514 nm/20 mw), HeNe (543 nm/1 mw, 633 nm/10 mw) - 6 channel AOTF - Excitation beam splitter: TD 488/543/633 nm; DD 458/514 nm, DD 488/543; RSP 500; grey filter: RT 30/70 - Conventional scanner: Up to 50 frames/s; line scan: Up to 2800 line/s - 1 TLD-detector for recording of transmission light images - 2 Internal detector channels for recording of fluorescent images - Objective: 5x/0.15 NA air; 10x/0.30 NA air; 20x 0.7 NA Mult. Imm.; 40x/1,3 NA oil; 63x/1,4 NA oil. A cell culture work bench HERAsafe, Thermo Scientific User Policy Core Facility LIMA Attachment 1: Technical Equipment Page 1 of 1

10 Attachment 2: Listing of user fees (November 2012) User fees are divided into 3 categories. Classification depends on the user s affiliation. Category 1: CBTM work groups and their staff, PhD and graduate students: 7,50 /hour Category 2: Members of the Medical Faculty Mannheim of Heidelberg University and other scientific institutions: 20 /hour Category 3: User from the industrial field: 100 /hour Night fee: A 50% reduction of user fees during the hours of 7 p.m. and 7 a.m. may be applied for long term experiments with a booking period of more than 6 hours. User training at the instruments will be charged in addition: Training of one user: 50 for the entire training session Training of 2 and more users: 30 /user for the entire training session Training fees will be charged starting January User Policy Core Facility LIMA Attachment 2: Listing of user fees Page 1 of 1

11 Attachment 3: Practical advices for the proper operation of instruments Before you start working: Check carefully the microscope and work place. If the work place is in an unacceptable condition (e.g. dirt), the instrument apparently does not run properly (e.g. misalignment etc.) or is damaged, you are obligated to inform a responsible person promptly and personally, at least via . Please report missing consumables, e.g. Kimwipes or immersion oil. Pay attention to the room temperature, LIMA rooms are air-conditioned. Please immediately report a noticeable increase higher than 26 C. During your work: If you are the first user of the day, turn on the fluorescent lamp, if needed. The fluorescent lamp should run for at least half an hour before being turned off. To prolong the fluorescent lamp life time keep the mechanical shutter closed as long as epifluorescent light is not needed. Activate only those laser lines in the software, which are finally needed for your probes and pay attention to the respective maximum value of the laser power. Unnecessary activation of the laser leads to its life-time shortage, even if the laser power is set to zero. For all lasers holds true: Laser needs to run for 2 hours after initialization. Turn off the laser if the subsequent booking is 2 hours later. If the subsequent booking is less than 2 hours then keep the laser running. Keep maximum power for the Ar + -laser at 20% as standard configuration. Higher laser settings e.g. for FRAP experiments require prior agreement. Carefully clean the objective and cover glass with a Kimwipe or Whatman lens paper soaked with isopropanol, if needed. Use the correct immersion medium (oil, glycerin, water, air) for your sample and select the appropriate objective. In addition, make sure the correction collar at the objective is adjusted properly. Please ask, if you are not sure! Please use only a small drop of the immersion medium and immediately remove excessive medium. Be concerned about the immersion medium and the availability of appropriate objectives before you start preparing your samples. User Policy Core Facility LIMA Attachment 3: Practical advices for the proper operation of the instruments Page 1 of 2

12 After you have finished your work: Clean the objective carefully and thoroughly with a Kimwipe or Whatman lens paper. In case you used glycerin or water take water for cleaning. Oil must be removed by isopropanol. If you are the last user of the day, shut down all systems and turn off the fluorescent lamp. After the fluorescence lamp has been turned off, it may not be turned on for at least one hour. Cooling of the laser has to run for at least 15 minutes before switching off. If you are registered as the last user of the day, implicitly remember to shut down the system and to turn off all instruments according to guidelines. This especially applies when cancelling your booking on short notice, since the preceding user does not know that you canceled your booking and keeps the system running. For the Multi Photon microscope: Remove the key from the microscope and keep in a code-locked key cabinet beside the entrance. User Policy Core Facility LIMA Attachment 3: Practical advices for the proper operation of the instruments Page 2 of 2

13 Attachment 4: Information about working at the S1 and with laser protection The rooms of the Core Facility LIMA s are security level 1 and are therefore only permitted for S1 samples. Here, the guidelines of the act on genetic engineering 12 paragraph 2 Gen TSV apply. Unsupervised work with hazardous substances is prohibited in LIMA s rooms. Instructions on working with genetically modified organisms (German GVOs) are within the responsibility of the user`s laboratory. Working with genetically modified organisms must be noted in the lab book by listing the species there. Working with infectious material (also donated blood) must be discussed with the persons in charge beforehand. Microscopes run class IIIb lasers (upright microscope) or IIIb and IV (inverse Multi Photon microscope). The participation at a laser safety instruction in accordance to accident prevention regulation Laserstrahlung BGV B2 is mandatory. Users are requested by the laser safety officer to participate at the general laser safety instruction once a year, as required by law. Each user is in charge of the microscopes lasers within the booked time period and is responsible for the proper and cautious operation of the lasers. LIMA s management will not take responsibility for improper laser operation and eventual consequent health damages. Modification and/or deflection of the beam path and therefore, of the laser beam, are principally not allowed. It is strictly prohibited to dismantle laser shields or laser coupling or even to open the laser cavity. Ensure that the laser s warning lights outside on the corridor are turned on when running the multi photon microscope. Always close the IR-laser s shutter when interrupting the experiment. User Policy Core Facility LIMA Attachment 4: Information about working at the S1 and with laser protection Page 1 of 1