Lane Lab

LSC USER Information

Return to: Cytometry Home Page Return to: Lane Lab Home Page

Item Last modified 6/1/01

The LSC Shared Instrument Faciltiy (LSC)

The Facility is under the direction of Dr. Timothy F. Lane (UCLA School of Medicine)

And is supported by a Shared Instrument Grant from the National Institutes of Health and

with contributions from the following::

The UCLA/Jonsson Comprehensive Cancer Center

The Provost's Office, and the Office of Research at the UCLA School of Medicine

The following UCLA Departments

Obstetrics and Gynecology,

Biological Chemistry,

Microbiolgy and Immunology,

Pathology and Laboratory Medicine,

Molecular Cellular and Developmental Biology


Table of Contents:

Statement of Purpose

Shared Use Policy Statement

Disclaimer

Data and Data Storage

Samples, Sample Storage, and Sample Disposal

Definitions

Biohazard Notice

User Responsibilities

Approvals

Appendix A:

Web accessable forms are still under development

Form A: Investigators Request for Use

Form B: Investigators Protocols

Form C: List of Samples and Reagents

Form D: Statement of Biohazards and protections

Form E: Letter from UCLA Biosafety Office

Appendix B:

Recharge Rates

Appendix C:

List of allowed and excluded substances

Statement of purpose:

The LSC_ Shared Instrument Facility (LSC-SIF) at UCLA is a cytometry facility that contains a Laser scanning cytometer. Because of very limited staff and resources, it is mainly intended for the use of the LSC Users Group. The LSC Users group is willing to support the development of LSC appropriate projects within the Jonsson Comprehensive Cancer Center and the UCLA research community.

Because of its very limited resources, the SIF does is not capable of providing service as a traditional core facility and will only accept samples on a case by case basis. Access to the facility will depend on approval by Dr. Lane, and on fluctuating requirements of the LSC Users Group.

We welcome collaborations, and will provide consultation and assistance with experimental design as time permits.

Shared Use Policy for the LSC:

The LSC SIF is not a full service core facility. The equipment is intended for the use of the LSC Users Group, and to assist when possible the development of LSC appropriate projects at UCLA. For full service cytometry, we encourage you to contact:

The Janis V. Giorgi Flow Cytometry Laboratory at UCLA

The LSC and all equipment in this lab belong to, and are controlled by Dr. Timothy F. Lane (Dr. Lane), UCLA School of Medicine. Dr. Lane is the final arbiter with respect to decisions about access and use.

The LSC is a very complicated mechanical device with a non trivial computer interface and certain inherent hazards (lasers and moving parts). It is not available for un-trained users. Training sessions can be organized with advance notice (see Recharge rates).

With proper notice and, in the event that it does not impair the progress of Dr. Lane's research, we welcome trained users (called "operators") to analyze appropriate samples on our equipment. No toxic or biohazardous samples can be analyzed (see relevant Biohazard sections of this manual).

It is our policy to provide access to the LSC during normal business hours. We ask all users to respect our time and to contribute to the cost of analysis through a recharge mechanism (see Recharge Rates).

Except for work with collaborators, we do not offer assistance with sample collection, preparation, or staining. In some cases, we can make suggestions about reagents and methods that have worked for others. These will only be suggestions and are not guaranteed to work for you (see Disclaimer).

You will be required to submit a simple application (Forms B and C) for use of the instrument (see user information). This process is not intended to be excessive but does provide us with information about the samples that will be used on our instrument. It also provides us with relevant contact information and billing information. In rare cases, you may be required to provide additional documentation (Forms D and E) and a letter from the UCLA Biosafety office demonstrating that your materials have been properly sealed and decontaminated (see biohazard requirements).

We can provide a Cytometry data sheet for each sample, but may be limited in terms of abilities to provide extensive support of your data analysis needs. You are encouraged to obtain copies of the software to allow more complete analysis of your data.

Data policy: You should maintain your own copies of all data. We can not be responsible for keeping backup copies. In some cases data may have been stored as part of our routine backup procedures. We will be happy to try to find your data if you can provide exact dates and sample numbers.

All users should be prepared to contribute data for future applications to support the upkeep, operation, and replacement, of these instruments.

All inquiries should be directed to: Dr. Tim Lane tlane@mednet.ucla.edu

Disclaimer:

All methods developed by the lab, or suggested for your use, are provided as a courtesy. They are only suggestions which have worked for some applications in the past. We do not guarantee that our methods will meet your needs, or work for your cells or application.

Data

    Data constitutes the results of the requested scan. The Operator will provide Data to the User in the form of:

        1. a print out of the results of the scan.
        2. on request: an e-mail will be sent with an attachment of the entire session file. The user is responsible for providing an active mail address and for insuring that the data was received and stored correctly.
        3. On request: as files on a computer disk (includes a disk fee)

Data Storage Policy

It is the Users responsibility to maintain copies of all primary data. Every effort will be made to store all data for one year on electronic media (Jazz Disks, CD ROM). Such storage can not be guaranteed and is subject to the many potential errors that can accumulate in electronic media. We will not store hard copies of the data. In some cases we may be able to retrieve data if provided with an exact date and a suitably informative Sample Identifier.

Sample:

Any single unit to be analyzed. An 8 chamber slide would have 8 samples. A Tissue microarray might have hundreds of samples.

Sample ID Number:

The Operator will assign each sample with a coded number by maintaining a log book with numerical codes. The user will provide brief notes on the sample (name, type, etc.). All information retrieval will be organized through this ID number.

Sample Storage and Disposal:

Samples will not be stored. Because of biohazards, we can not dispose of your samples. Users are responsible for disposing of all samples. Failure to pick up your samples in a timely fashion could affect your ability to use the facility in the future.

Definitions:

SIF = the LSC_ Shared Instrument Facility

LSC Users Group:

PI email Project:
LANE, TF tlane@mednet.ucla.edu Quantitation of Nuclear Translocation and Functional Cloning of Target Genes in Breast and Ovarian Cancer
Bonavida, B bbonavida@mednet.ucla.edu Analysis of NK and CTL cell populations and the sensitization of tumor cells to TNFa-mediated apoptosis
Chaudhuri, G gchaudhuri@mednet.ucla.edu Analysis of Nitric Oxide (NO)-Induced Cell Cycle Arrest in Human Breast Cancer Cells
Herschman, H hherschman@mednet.ucla.edu Primary response gene expression in neuronal plasticity and neuronal differentiation
Iruela-Arispe, L arispe@mbi.ucla.edu Effect of angiogenesis inhibitors on endothelial cytoskeleton
Rao, J jrao@mednet.ucla.edu Cellular G/F-Actin Ratio as a Marker for Malignant Transformation and Apoptosis
Rome, L lrome@mednet.ucla.edu Quantitation of Vault Expression in Breast Cancer to Predict Drug Resistance and Prognosis
Zack, J jzack@mednet.ucla.edu Analysis of complex interactions in the human thymus

Investigator = Principle Investigators at UCLA or at other institutions who are requesting work or reagents from the SIF. This person is responsible for all work being requested and for authorizing all Users. The Investigator is also the person responsible for establishing safe laboratory practices for each sample type, guaranteeing that samples display no hazard to the operator or the instruments, and for paying for services incurred by the Users.

Users: = Any researcher (faculty, student, technician, or other) who brings samples to the facility.

This person will be approved by the Investigator to request work and reagents from the facility. This person will log each sample in to the log book and must be able to explain the identity of the sample, the type of analysis needed, the presence of any hazardous agents, and the measures taken to eliminate exposure to such agents.

Operator(s): = a subset of users who are trained to operate the instrument. They can run samples and provide data on the sample.

Sample: Any single unit to be analyzed. An 8 chamber slide would have 8 samples. A Tissue microarray might have hundreds of samples. (see Samples and Sample Storage)

Sample ID Number: The Operator will assign each sample with a coded number by maintaining a log book with numerical codes.

Log Book = a Laboratory Notebook which provides the site for entry of all primary information from the User related to each sample.

Data =the results of the requested scan. The Operator will provide Data to the User. (See Data and Data Storage)

Biohazard and Sample Responsibility Notice

The nature of microscope based experiments requires some amount of handling of each sample/slide by the microscopic technician/operator. Improperly prepared samples can pose a risk to personnel AND to the equipment. Do not assume, that the operator will be able to avoid non-routine hazards in your sample. It is your responsibility (the user/investigator) to seal and clean all contact surfaces from your samples so that there is no risk of exposure to the technicians OR to the instrument. Sloppy/leaky samples will be rejected and should not be presented to the lab (see User Responsibility Rules).

Biohazard Notice:

All samples/projects must be PRE-approved (see below) for use on these instruments.

No samples containing restricted biohazardous agents (level II or greater) can be analyzed or brought to the Lab without appropriate decontamination, and/or foolproof measures to restrict exposure.

No infectious, caustic, mutagenic or fixative agents can be used in such a manor that exposes personnel or equipment in this facility.

The samples must not pose a biohazard to personnel.

The samples must not pose a mechanical or chemical hazard to the personnel or instrumentation.

All samples must be presented in a fashion that would allow our technician to pick up the sample carrier (e.g. the glass slide) without the risk of biohazard or mechanical exposure.

This includes (but is not limited to): No: sharp edges, infections agents, caustic agents, undiluted fixatives, or noxious fumes, etc.

In most cases, this will require a cover slip and sealant so that fluids can not leak onto hands or surfaces

User Responsibility Rules:

    The user (i.e., anyone dropping off samples), is responsible for:

        1. knowing and understanding the types of agents that are contained in the samples and that their condition meets or exceeds safety requirements.
        2. informing our staff of sample contents and safety issues.

    Lack of such knowledge may cause exclusion of the sample from the facility and other actions.

Approvals:

For all work in the Lane Lab and the LSC Shared Instrument Facility, we must have on file:

    1. A statement of Approval for your project, provided by Dr. Lane,
    2. A letter from the Investigator (UCLA PI) requesting the use of the facility, agreeing to abide by it's rules (which may change without notice), and agreeing to pay for all services as specified in our Recharge Rates Announcement (which may change without notice) (see FORM A)
    3. For all non LSC-User group labs, a protocol explaining the goals or the project and describing how the work will be funded. This should include an explanation of why the work can't be done in the Janis V. Giorgi Flow Cytometry Laboratory, which is the more appropriate venue for most fee-for-service cytometry at UCLA. (see FORM B)
    4. a statement as to the nature of the samples (cell types, human/mouse) and all agents with particular mention of potential risks and biohazards. (See FORM C)

Additional Requirements when samples contain toxic, caustic, or infectious agents (viruses, chemicals, etc.)

V. a statement as to how the samples have been detoxified or how they have been sealed to prevent exposure of personnel AND equipment. (See FORM D)

VI. a letter from the UCLA biosafety office approving the work and referring specifically to the use of these samples on the LSC or other equipment in the Lane Lab. This is required for any sample exposed to a human infectious agent such as HIV/AIDS, Herpes, etc. (see FORM E).

 

LSC Shared Instrument Facility:

Appendix B:

Laboratory Charges

Operator-conducted Acquisition or Analysis (1hour Minimum. Thereafter, time calculated to the nearest 0.5 hour)

LSC USERS GROUP $60.00/hour

Members of LSC supporting Departments $70.00/hour

Other UCLA Users $90.00/hour

Lane Lab Operator $90.00/hour

Non-UCLA Affiliates (reviewed on a case-by-case basis)

Corporate Rates (reviewed on a case-by-case basis)

Staining and Analysis (2 sample minimum)

In general, the SIF does not provide reagents, slides, or staining services.

Single Color $20.00/sample

Dual Color $40.00/sample

Triple Color $60.00/sample

Slides/cover slips $1.00/each ($20/box)

Antibodies/Reagents (Stains > 5x105 Cells) $5.00/aliquot

Disks

650 MB Recordable Compact Disk (CD-R) $10.00/Disk

100 MB Zip Disk $20.00/Disk

Floppy diskette (High density)2.MB $5.00/Disk

Instructional Sessions

Analytic Cytometry Instructional Course (4 hours) $440.00

 
Appendix C:

List of Allowed and Excluded Substances

Allowed Substances

Most standard cytometry and flow cytometry reagents [DAPI, PI, etc]

Excluded Substances

No live/active human pathogenic material of any kind (viruses, bacteria, etc.). All material must be fixed and inactivated to the satisfaction of the UCLA BioSafety Officer and Dr. Lane.

Caustic, Sharp, or Volatile agents which can injure the operator or damage the microscope, computer, or other equipment..

Any agent requiring spicific Approval of the Environmental Health and Safety Office

Please contact the UCLA BioSafety Office or Dr. Lane for verification

   

Return to:[top]

Return to: [Home]