Applications are developing in localization microscopy where the number of targets for imaging far outstrips the number of probes that can be chromatically separated. Sequential labelling with microfluidics allows a much larger number of targets to be addressed. Bruker microfluidics accessory units are designed to allow sequential labelling to be incorporated into localization microscopy protocols with the Vutara VXL single-molecule localization microscope.
Bruker offers two units—PlexFlo and PlexFlo96. The units are controlled by Vutara's SRX software and allow flexible configuration of fluidics sequences that are run in conjunction with imaging cycles to allow users to label an unlimited number of probes and image them in a sequential fashion.
PlexFlo contains 15 buffer reservoirs that can be swapped out during an experimental workflow, enabling the multiplexing of many labels in one experiment. These distinct features make PlexFlo optimal for DNA-PAINT applications.
PlexFlo96 enables even further multiplexing capabilities, as it contains 96 buffer reservoirs that can be swapped during one experiment. Consequently, PlexFlo96 is ideal for applications that require a large number of labels in one experiment, such as chromatin tracing.
The probe associated with each sequence has a specific color that is user-defined, and that is used as a color scale for visualization of experiments. During analysis with SRX, individual probes can be analyzed, or analysis can be carried out that compares probes.
The Bruker microfluidics unit can be used with Vutara to address any localization microscopy application where a large number of targets needs to be imaged. Fifteen probe vials are provided, and the system allows probe vials to be interchanged during experiments, thus allowing an unlimited number of targets.
Example localization microscopy applications:
The Bruker microfluidics unit is also compatible with the Opterra confocal for serial labelling confocal applications.
|Samples/Reservoirs (small volume)||15 x 2 ml vials (Eppendorf standard)||96 well in microwell plate|
|Wash/Buffer Reservoirs (large volume)||1 x 100 ml, 1 x 50 ml, 2 x 15 ml||8 x 125 ml|
|Waste Container||1 x 100 ml GL45 bottle|
|Temperature Control of Sample||No||39￮ - 99￮F; 4￮ - 37￮C (microwell plate)|
|Sample Swap During Experiment||Yes||Yes|
|Pressure Driven Flow Control||Yes||Yes, and vacuum for extraction from 96 well plate|
|Pressure Range||0 – 345 mbar (±0.1% CV above 10% full scale, ± 105 µbar resolution)|
|Flow Sensor||Thermal based, 1 µl/min to 1 ml/min (±1% CV)|
|Automatic Cleaning Procedure||Yes||Yes|
|Dimensions||370 mm x 300 mm x 204 mm|
Control of the unit is via SRX software. The software has a flexible interface module (Fluidics Manager) for the set-up of fluidics. An unlimited number of sequences can be set up, each with its own unique steps.
The Bruker microfluidics accessory is a unit designed to allow sequential labelling to be incorporated into localization microscopy protocols with the Vutara VXL single-molecule localization microscope. The unit is controlled by Vutara's SRX software and allows flexible configuration of fluidics sequences that are run in conjunction with imaging cycles to allow users to label an unlimited number of probes and image them in a sequential fashion.