Malaghan Institute of Medical Research

The Malaghan Institute Flow Cytometry Suite provides access to the following resources:

Benchtop Analysers: FACSort, FACSCalibur, LSRII

Cell Sorter: FACSVantage DiVa

Ancillary Equipment: High Throughput Sampler for LSRII, BioPlex, AutoMACS

The Sample Injection Probes (SIPs) of our flow cytometers are compatible with 12x75mm, 5mL polystyrene round bottom test tubes and the recommended cell concentration is two million cells per mL, resuspended in a minimum volume of 500µl.

It is important to note that dedicated operator assistance is only provided for the flow sorting equipment - all researchers will be expected to run the analysers unassisted after the initial training session.

If you require further assistance after your initial training please ensure you contact a Flow Suite staff member to arrange a time PRIOR to your booking.

 BD FACSort and BD FACSCalibur

These two benchtop analysers are capable of supporting a wide range of basic flow cytometric applications. They have two lasers (blue, 488nm and red, 633nm) and are capable of detecting four colours. These two benchtop analysers are engineered with fixed optical, electronic and fluidic components, giving them the flexibility to function as an investigator-operated instrument, unlike the cell sorter.

 BD LSRII Special Order Research Product (SORP)

The LSRII flow cytometer is the only digital bench top analyser flow cytometer in the facility. It is an 18-colour, 5-laser machine that has optical detectors and removable optical filters that can permit the detection of different fluorophores and fluorophore combinations.

The 18 detectors are spread out over 3 Trigons (off the UV, Blue and Red lasers) and 2 Octagons (off the Violet and Green lasers) - refer to LSRII SORP Detector Configuration table at the bottom of this page for a complete list of bandpass filters.

The 5 lasers are as follows:
1.   355nm, 20mW, UV
2.   405nm, 50mW, Violet
3.   488nm, 100mW, Blue
4.   532nm, 150mW,Green
5.   640nm, 40mW, Red

 BD FACSVantage DiVa

The FACSVantage DiVa is a high performance, high speed, sterile, cell sorter that can isolate up to 15,000 individual cells per second. Sorted cells are then returned to the researcher who continues their study into how this cell of interest may be involved in the particular disease they are studying. The sorter is capable of simultaneously sorting up to 4 different cells from a given sample.

The cell sorter has 3 lasers (355nm, 100mW Nd:YAG UV laser, 488nm 100mW Argon ion blue laser and a 633nm, 35mW Helium-Neon red laser) and 7 possible detectors, listed below:

Unlike the bench top analysers, which are operated by the investigators themselves, sorter operation is provided as a service (see Facility Rates). This is because the instruments are much more complex than the bench top analysers and requires special training and a vast amount of experience to achieve the desirable results.

Because the sorter uses a stream-in-air sorting method, it aerosolises the samples. Consequently, the sorters cannot be used for sorting hazardous samples that require PC3 containment. However the sorter is equipped with an aerosol containment unit and a Personal Air-Purifying Respirator (PAPR), therefore some special accommodations can be made for potentially infectious sorts (up to PC2 level), please enquire in advance. Sort clients must complete the Biosafety Assessment - Cell Sorting Log Sheet in order to assess the risk level of the sort, before the sort can be accepted and performed.  Please contact a Flow Cytometry Suite staff member for a copy of this Log Sheet.

  BD High Throughput Sampler (HTS) for the LSRII

The HTS option allows users to rapidly acquire samples from plates, the HTS supports 96-well U-, V-, and flat bottom plates, as well at 384-well microtiter plates. BD reports carryover tolerances of <1% even during high-speed acquisition.

There are two throughput modes: Standard and High-Throughput and the recommended sample concentration is 1x107 cells/ml. The HTS can process a 96-well plate in ~45 minutes in "Standard" mode (taking 10µl of sample) and ~15 minutes in "High-Throughput" mode (taking 1µl of sample).

In "Standard" mode you can take anywhere between 10-200µl of your sample, so between 100,000 and 2x106 cells per well at the recommended concentration. And in "High-Throughput" mode you can take anywhere between 1-10µl, so between 10,000 and 100,000 cells at the recommended concentration.

In the flow cytometry analysis software FlowJo, it is simple to concatenate files (join FCS files together), so you can run one sample over several wells to acquire your target number of events, if you require more than 2x106 events (in standard mode) or more than 100,000 events in "High-Throughput" mode.

 BioRad BioPlex 200 – Multiplexing Bead Array Plate

A 96-well plate-based multiplexing platform that allows for simultaneous quantitative analysis of up to 100 different analytes/proteins in a single well.

The BioPlex 200 is built on xMAP® technology and is remarkably easy to use, it combines 2 lasers, fluidics, and real-time digital signal processing to distinguish up to 100 different colour-coded bead sets, each representing a different assay. The microplate platform automates the reading of 96-well plates and yields up to 9,600 data points in ~35 minutes. Our BioPlex is also equipped with the High-Throughput Fluidics (HTF) option which delivers up to 20L of sheath fluid (~40 plates) without user intervention.

A number of companies produce or provide xMAP® technology-compatible kits that can be run on the BioRad BioPlex 200:

 AutoMACS

The autoMACSTM Separator is a benchtop instrument for high-speed automated cell sorting. Employing the MACS® Technology, the autoMACS Separator is designed for positive selection as well as depletion of magnetically labelled cells.

The autoMACS Separator is operated with preset separation programs, thus, allowing optimisation of cell sorting approaches according to cell abundance and the intensity of marker expression.

The separated cells are immediately ready for experiments, cell analysis, or further subset sorting. Automated system cleaning allows convenient maintenance.