The EnVision® microplate reader has capabilities across various measurement technologies including absorbance, fluorescence intensity, fluorescence polarisation (FP), time-resolved fluorescence (TRF), luminescence and AlphaScreen. For automated high-throughput screening, the EnVision is serviced by a Denso Plate Handler 6-axis robot, which is capable of moving plates out of a microplate hotel (at ambient temperature), a temperature-controlled incubator, or from other pieces of equipment into/out of the EnVision instrument.

Technical Specifications

For absorbance and top-read fluorescence intensity measurements, the instrument features a monochromator that has the following ranges:

• Absorbance: 230 – 1000 nm
• Fluorescence intensity: 230 – 850 nm

For filter-based fluorescence intensity, FP, TRF and AlphaScreen measurements we currently have the following filters:

• 405/8
• 531/25
• 535/40
• 570/100 (used predominantly for AlphaScreen)
• 579/25
• 615/5
• 642/5

If your assay requires fluorescence intensity, FP, TRF or AlphaScreen measurements at wavelengths outside of these filter ranges, please consult directly with ACTD staff.

Assay Optimisation on the EnVision

Prior to running an experiment for the first time on the EnVision, a series of optimisation steps must be completed in order to maximise the results obtained.

• Plate dimensions: Clients should provide ACTD staff with a sample plate no less than 24 hours prior to their first appointment. Please also attach information on the plate type and catalogue number (a plate specification sheet would also be ideal).
• Measurement height: In the first instance, clients should provide at least 3 negative control wells and 3 positive control wells (with a strong and well-developed signal) within their plate on the day of the experiment.
• Detector gain (only for fluorescence intensity or FP): As above, clients should provide at least 3 wells with a strong, well-developed signal on the day of the experiment.

Plate Type

Often the plate type chosen will be specific for a given assay and the type of measurement technology to be used. Please keep in mind the following considerations when choosing a plate type:

• Opaque-walled plates are recommended to prevent signal cross-talk between wells when reading fluorescence.
• White plates reflect light resulting in higher signals whereas black plates tend to quench light. However, black plates are generally recommended for fluorescence assays as they provide a higher signal-to-noise value and reduce background autofluorescence.
• Time-resolved fluorescence assays use longer half-life fluorophores and incorporate a lag or delay time after excitation, therefore diminishing background autofluorescence. White plates are usually recommended for TRF applications.
• For cell-based assays that require culturing cells in an assay plate over multiple days or weeks, sterile tissue culture-treated (TC) plates are recommended.
• For AlphaScreen assays PerkinElmer AlphaPlates (light grey) are recommended. Black plates are generally not recommended for AlphaScreen assays.

Many of the plate types described above can be purchased from the ACTD supply store – please chat to the ACTD staff for more information.

Data Management

Data generated on the EnVision will be sent to the client either via an email from ACTD staff or for larger files through ANU CloudStor. It is the responsibility of the client to save all important data in a secure networked folder or hard drive and to make back-ups of this data regularly. The ACTD will store the raw EnVision data files for a period of one month before deleting them. As this temporary storage is hosted by the ANU network, the ACTD cannot guarantee this data and it is recommended that clients save, store and make a copy of their data as soon as it is sent to them by staff.