Acumen Technology

Acumen ^eX3 Microplate Cytometer Technology
Acumen ^eX3 uses multiple scanning lasers (at 405, 488 and 633 nm) to excite fluorescent objects on the bottom of microplates. The lasers fire sequentially to reduce crosstalk and give simultaneous detection in four photomultiplier tube (PMT) detectors, giving a maximum of 12 channels of data for true multiplexing at high throughputs. Acumen's maximum scanning resolution equates to that achieved using a 20x microscope objective.

Whole well scanning Acumen ^eX3 has a specialised scan lens with a very wide field of view of 400mm² (20 x 20mm or equivalent to 4 wells of a 96-well plate). This enables Acumen to scan entire wells and report data for all the cells to: give statistically robust data from a truly representative cell population - essential for rare event detection assays. offers normalisation of biological responses to total cell number. removes the need for image stitching for analysis of large objects such as C.elegans, tissue sections or cell colonies. Whole well scanning is not feasible for microscope-based imaging technologies due to their much smaller fields of view - typically <1mm² for a x10 objective, which equates to 49 fields of view per 96-well.	Using a x10 objective, an imaging system's field of view equates to 2.6% of a well in a 96-well plate

Cytometric analysis Acumen's lasers scan the entire field of view collecting fluoresence intensity readings at regular intervals using 4 PMTs. Thresholding algorithms identify all fluorescent intensities above the solution background for automatic object identification. Fluorescence detected by each PMT is displayed as 3-dimensional object intensity profiles. These profiles enable the calculation of a range of morphological and fluorescent parameters. Laser scanning and cytometric analysis mean that Acumen ^eX3 gives extremely fast results - averaging around 10 minutes per plate.	Blue lines indicate laser scanning across a single cell, circles represent intensity readings

Data storage Data from the Acumen ^eX3 can be collected in 3 levels of detail to suit the assay development, assay validation and screening scientist. Even during assay development files rarely exceed 350 MB. In high throughput screening mode, the software reports only population statistics, reducing file sizes down to a manageable 50 kB.