UCL Cancer Institute

Flow Cytometry

We have two systems for Flow Cytometry. Both allow the user to measure simultaneously multiple physical characteristics of a single particle (cell) in a moving fluid stream:

• The Cyan ADP analyses the relative size, internal complexity and fluorescence of a cell.
• The MoFlo is a high speed 4-way sorter which can measure up to 14 parameters.

Flow Cytometry - CyAn ADP

The CyAn ADP High-Performance Flow Cytometer is a state-of-the-art flow cytometer that utilizes multiple laser excitation sources to analyze biological cells, beads, or other microscopic particles. It remains a closed fluidic system – also allowing analysis of biohazardous samples (such as human blood samples, bacteria, etc), with appropriate precautions and authorization. It provides excitation with 405nm, 488nm and 633 nm lasers – with 9 detection channels providing a broader range of detection parameters in tailored to accommodate a wide range of experiments. This, combined with the system’s optics, provides enhanced performance for all applications including dim cell populations. High sensitivity and greater resolution for each parameter give you accurate results whether you are running a three color application or taking advantage of the nine fluorescence channels.

The CyAn ADP has cutting edge optics, utilizes high performance digitization with a pulse processing speed of up to 70,000 events per second (complementing a high acquisition) and analysis capability of up to 100,000,000 events. Typically, investigators acquire 5,000 to 100,000 cells per sample.

Up to 9 x 9 channel compensation can be performed – during (online) or after (offline) running sample controls – either manually, using bi-exponential transformation (VisiComp), or automatically.

Summit Software is easy to learn and operate. It offers a workspace concept, detachable menus, drag and drop mouse functions, as well as data presentation tools such as overlay plots and multi file plots. The comprehensive Graphical User Interphase (GUI) is convenient both for online and offline work. Data can be saved to the hard disk of a dedicated PC computer and can later be analyzed with the associated graphics software, Summit v4.3. Data can also be transferred via a USB to any other computer convenient to the user and analyzed with a copy of the Summit software, as well as Flow Jo.

It is easy to use and can be operated by the experimenters themselves - after the prerequisite course of training by our technicians. When attempting to use new and unfamiliar protocols please ask one of the technicians to clarify any uncertainties.

Please note that users must run their own samples after prerequisite session of training.

Measurements are made on a cell-by-cell basis so it is important that samples are prepared in a single cell suspension to reduce the occurrence of blockages. A full cleaning protocol must be carried out at the end of each use so that it is in a useable condition for the next person.

Training

All prospective users must have a training session prior to running there own samples.

Booking

Wolfson and Cancer Institute staff please book Via WORMS, other UCL staff and external users please contact us for booking.

Flow Cytometry - MoFlo

The MoFlo is an air-in-jet high-speed cell sorter, equipped with three lasers that can measure up to 14 parameters and sort 4 populations simultaneously. It is fitted with a CyClone, the automated deposition unit, allowing single cell deposition for cloning purposes. Sorting under sterile conditions can be done if requested and sorted cells can be collected in several various vessels (e.g. 1.5-50ml tubes, as well as 6-96-well and 10cm well plates).The MoFlo temperature can be controlled over the sorting time from 4-37°C. It can be set up with different nozzle sizes - from 50-150um - which allows a nice range of cell sizes to be analyzed. The MoFlo typically sorts at rates of 10,000 - 30,000 cells/second, with over 95% purity and good recovery rates. The sorting mode can be altered to give maximum purity, maximum recovery or maximum count accuracy - depending on the request. Due to the generation of aerosols, a risk assessment will be performed before accepting any samples.

The data can be analysed by the operator and given to you as a "ppt" file, or you may request a FCS file so you can do further analysis yourself, with the appropriate software (e.g. WinList, FlowJo, WinMdi). If you don't have any of these: please ask.

Applications

• Cell Cycle analysis
• Apoptosis analysis
• Multicolour analysis
• Functional analysis
• Cell Sorting

Booking

Access is at present allocated with equal priority to WIBR users (3 days a week) and the Windeyer Institute (2 days a week). Other users will be allocated time around these prioritised users, if available. Please contact Tomas for further information.

Prices: