Welcome to the HSCI-CRM Flow Cytometry Core Facility!
The CRM Flow Cytometry Core Facility offers dedicated research space, specialized equipment, and trained technicians to assist researchers with their cytometry sorting and analysis needs. Hours of Operation: Monday-Tuesday: 8:00AM-6:30PM; Wednesday - Friday 8:00AM-5:00PM Staff: Maris Handley, Nafeez Ishmam Ahmed, and Luis Orozco Phone: 617-643-0790 Email: mghcrmflowcore@mgb.org Location: Simches 4310
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You need to know!! (last updated 9/17/24): Flowjo portal licenses need to be renewed as of 9/15/24. The link is here: https://rc.partners.org/kb/article/1459 9/2/24: Aria 2 is down. Currently, we don't know when it will be available again. The Bigfoot schedule has been opened with the same hours, so sort availability isn't impacted for staff assisted sorting. We apologize to the self-run users for loss of available time 10/1/24: We will no longer be training new Aria users. If you have already started training, you are welcome to finish. If you are collecting data for analysis, you may train on the Cytpix. If you are sorting, you may train on the Bigfoot. Ongoing: The first Tuesday morning of each month is blocked off for instrument and facility cleaning; no instruments are available during this time. Holidays and facility closures (self-run users still have 24/7 access) Oct 14th: MGH holiday Nov 28-29th: Thanksgiving and following Friday Dec 25th: Christmas (other days TBD) Jan 1st: New Years Day Upcoming CytPix classes: Coming soon
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New? Start here:
Prior to scheduling time at our facility, you will need to 1. create an account on our online system, 2. link a fund number to that account, and 3. Complete a Biosafety Agreement. Biosafety Agreement – For all new users. Please complete this form if you intend to use our core facility. This needs to be submitted to us before scheduling any time. Administrative and Experimental Info Sheet — This sheet will provide you with all of the necessary information you will need prior to using our core. We are happy to answer any questions you may have! Pricing information for each of our services and equipment can be found here Interested in training on one of our machines? Click here
Sorting Human Cells:
We are now able to run a wider variety of BSL2 samples on our Aria3 and Bigfoot sorters. Per Universal Precautions: All primary human samples will be considered BSL2 samples. Prior to being able to run such samples, please complete the following questionnaire: BSL2+ PI Questionnaire - To be filled out by the PI of any lab that wishes to sort BSL2 samples. Our BSL2+ sorting policies can be found here. Please review them prior to sorting BSL2 samples with us. The Amendment form can be found here.
Note: High fluid pressures required in high-speed cell sorters can generate large amounts of aerosols of various and undefined droplet sizes during instrument failures, for instance, when a partial clog in the nozzle causes a deflection in the fluid stream that is hitting a hard surface, e.g., the waste catcher. The HSCI-CRM flow core provides sorters with varied levels of aerosol mitigation to protect instrument operators and anyone in the vicinity of the sorter. The level of containment is based on risk level. The most basic containment is provided by enclosing the stream within a sealed chamber, and the sort receptacles which are connected to this closed chamber have no gaps that would allow aerosols to escape. This is containment is suitable for cells that pose a very low risk. This might include murine cells that are uninfected, or fixed samples. For samples that are BSL1, for example BSL1 human cell lines, or cells infected with non-replicating viruses for transfection, the closed chamber has a vacuum applied via an Aerosol Management System (AMS) which actively removes any stray aerosols. The sorter is enclosed in a biosafety cabinet for additional operator safety. For BSL2 samples – any primary human cells, non-human primate cells, or infected samples – the sorter is enclosed in a biosafety cabinet with an AMS; it has also been placed in a separate room to limit access while the sort is happening.
Click on the links below to learn more about each of our instruments:
Aria 1 — FACSAriaIIu in hood Aria 3 — FACSAriaII in hood Bigfoot MACSQuant VYB Cytpix
Spectraviewers/Panel Design:
https://www.bio-rad-antibodies.com/spectraviewer.html https://www.biolegend.com/spectraanalyzer https://www.thermofisher.com/order/fluorescence-spectraviewer#!/ Fluorescent proteins Fluorofinder (our default configurations are here) https://www.thermofisher.com/order/panel-builder/#!/
Forms:
Biosafety Agreement – For all new users. Please complete and submit this form if you intend to use our core facility. This needs to be submitted to us before scheduling any time.
Human Sort Document – For anybody bringing live, tested, uninfected human samples (non-BSL2). You must either bring this form every time you bring live human cells, or link to fill out the online form here.
Anybody working with live human cells (non-BSL2 cell lines) MUST also bring documentation stating that the cells have been tested and found negative for HIV, HepB, and HepC and are otherwise non-infectious samples.
BSL2+ PI Questionnaire - To be filled out by the PI of anyone who wishes to sort BSL2 samples.
BSL2+ Amendment Form - For minor updates to the BSL2+ form
Other useful information: Introduction to Flow Cytometry – an excellent collection of flow cytometry training videos provided by BD Biosciences FlowBook - Intro to Flow - created by DeNovo Software, another great resource for learning about flow cytometry Compensation Tutorial – courtesy of Dr. Mario Roederer, this website will teach you about compensation Intro to Spectral Flow - educational videos from BD Biosciences Cytek's SpectroLearn - educational videos from Cytek Thermofisher videos about the Cytpix - videos to help getting started on the CytPix Data Analysis - Part 1 (uses FlowJo); Part 2 (uses FCSExpress) New England Cytometry – stay up to date on all of the flow cytometry happenings in the New England area
